Diel cycles of carbon, nutrient and metal in humic lakes of permafrost peatlands.

Despite the importance of surface waters of permafrost landscapes in carbon (C) emission and dissolved C and metal storage and export, the majority of available observations in high latitude aquatic systems deal with punctual or seasonal sampling without accounting for diurnal variations in temperature and primary productivity-respiration cycles. Towards providing comprehensive understanding of diel variations in CO2 emission, organic C and element concentrations in lakes of frozen peatlands, we monitored, each 2 h over 2 days, the water temperature, pH, CO2 fluxes, CO2, CH4, dissolved organic and inorganic carbon (DOC and DIC, respectively), nutrients, carboxylic acids, bacterial number, and major and trace elements in two acidic (pH = 3.6 and 4.0) and humic (DOC = 15 and 35 mg L-1) thermokarst lakes of discontinuous permafrost zone in Western Siberia. We discovered a factor of 2 to 3 higher CO2 concentrations and fluxes during the night compared to daytime in the high-DOC lake. The emission fluxes in the low-DOC lake increased from zero to negative values during the day to highly positive values during the end of night and early morning. The methane concentration varied within a factor of 5 without any link to the diurnal cycle. The bulk of dissolved (< 0.45 µm) hydrochemical parameters remained highly stable with ±10% variation in concentration over 2 days of observation (DOC, DIC, SUVA254nm, carboxylates (formate, oxalate, puryvate and glutarate), Mn, Fe, Al, other trace elements). Concentrations of Si, P, K, Cu varied within ±20% whereas those of Zn and Ni ranged by a factor of 2 to 4 without any link to diurnal pattern. Overall, the impact of diel cycle on CH4, DOC, nutrient and metal concentration was below 10%. However, neglecting night-time period may underestimate net CO2 emission by ca. 30 to 50% in small organic-rich thaw ponds and switch the CO2 exchange from uptake/zero to net emission in larger thermokarst lakes. Given the dominance of large lakes in permafrost regions, the global underestimation of the emission flux may be quite high. As such, monitoring CO2 concentrations and fluxes in thermokarst lakes during months of extended night time (August to October) is mandatory for assessing the net emissions from lentic waters of frozen peatlands.

Contrasted resistance and resilience of two mangrove forests after exposure to long-term and short-term anthropic disturbances.

Mangroves, coastal forests under the influence of tides, are known to be very resilient when they face natural disturbances such as storms or tsunami. While they provide several ecological services, they are threatened by many anthropic pressures. The aim of this study was to assess and to compare the stability of two mangrove fringes defined by contrasted set of natural constraints and exposed to pretreated domestic wastewaters discharges. The in situ experimental system set up in Mayotte Island (Indian Ocean) allowed us to determine both the short-term (2 years) and the long-term (9 years) resistance and the resilience. We focused on vegetation and crabs, an essential component of mangroves fauna. Wastewater discharges induced increases in tree coverage, leaves productivity and pigment content, and a decrease in crab diversity and density. Within 2 years after the release of the disturbance, several parameters reach back control values indicating fast resilience. Our results notably emphasized the high stability of the mangrove fringe dominated by Rhizophora mucronata trees, which was both more resistant and more resilient. This makes this fringe more suitable for application purposes, such as outfall for domestic wastewaters treatment plants.

Limited impact of several years of pretreated wastewater discharge on fauna and vegetation in a mangrove ecosystem.

It was hypothesized that mangroves, tropical wetlands, could be used for the finishing treatment of domestic wastewaters. Our aim was to determine if a nutrient-stressed mangrove could tolerate long-term discharges of pretreated wastewater (PW). Since 2008, in an in situ experimental system set up in Mayotte Island (Indian Ocean), domestic PW are discharged into two impacted areas (675 m2) dominated by different species of mangrove trees. Anthropogenic inputs during > 4.5 years led to an increase in vegetation growth associated with an increase in leaf pigment content, leaf surface and tree productivity. A marked increase in tree mortality was observed. There was no effect on crabs and meiofauna densities, but significant modifications of community structures. These effects may be directly linked to PW inputs, or indirectly to the modifications of the environment associated with higher tree growth. However, our results indicate that there was no major dysfunction the ecosystem.

Response of three biofilm-forming benthic microorganisms to Ag nanoparticles and Ag+: the diatom Nitzschia palea, the green alga Uronema confervicolum and the cyanobacteria Leptolyngbya sp.

Although the industrial use of nanoparticles has increased over the past decade, the knowledge about their interaction with benthic phototrophic microorganisms in the environment is still limited. This study aims to characterize the toxic effect of ionic Ag+ and Ag nanoparticles (citrate-coated silver nanoparticles, AgNPs) in a wide concentration range (from 1 to 1000 µg L-1) and duration of exposure (2, 5 and 14 days) on three biofilm-forming benthic microorganisms: diatom Nitzschia palea, green algae Uronema confervicolum and cyanobacteria Leptolyngbya sp. Ag+ has a significant effect on the growth of all three species at low concentrations (1-10 µg L-1), whereas the inhibitory effect of AgNPs was only observed at 1000 µg L-1 and solely after 2 days of exposure. The inhibitory effect of both Ag+ and AgNPs decreased in the course of the experiments from 2 to 14 days, which can be explained by the progressive excretion of the exopolysaccharides and dissolved organic carbon by the microorganisms, thus allowing them to alleviate the toxic effects of aqueous silver. The lower impact of AgNPs on cells compared to Ag+ can be explained in terms of availability, internalization, reactive oxygen species production, dissolved silver concentration and agglomeration of AgNPs. The duration of exposure to Ag+ and AgNPs stress is a fundamental parameter controlling the bioaccumulation and detoxification in benthic phototrophic microorganisms.

Comparative responses of river biofilms at the community level to common organic solvent and herbicide exposure.

Residual pesticides applied to crops migrate from agricultural lands to surface and ground waters. River biofilms are the first aquatic non-target organisms which interact with pesticides. Therefore, ecotoxicological experiments were performed at laboratory scale under controlled conditions to investigate the community-level responses of river biofilms to a chloroacetanilide herbicide (alachlor) and organic solvent (methanol) exposure through the development referenced to control. Triplicate rotating annular bioreactors, inoculated with river water, were used to cultivate river biofilms under the influence of 1 and 10 µg L(-1) of alachlor and 25 mg L(-1) of methanol. For this purpose, functional (thymidine incorporation and carbon utilization spectra) and structural responses of microbial communities were assessed after 5 weeks of development. Structural aspects included biomass (chlorophyll a, confocal laser scanning microscopy) and composition (fluor-conjugated lectin binding, molecular fingerprinting, and diatom species composition). The addition of alachlor resulted in a significant reduction of bacterial biomass at 1 µg L(-1), whereas at 10 µg L(-1), it induced a significant reduction of exopolymer lectin binding, algal, bacterial, and cyanobacterial biomass. However, there were no changes in biofilm thickness or thymidine incorporation. No significant difference between the bacterial community structures of control and alachlor-treated biofilms was revealed by terminal restriction fragment length polymorphism (T-RFLP) analyses. However, the methanol-treated bacterial communities appeared different from control and alachlor-treated communities. Moreover, methanol treatment resulted in an increase of bacterial biomass and thymidine incorporation as well. Changes in dominant lectin binding suggested changes in the exopolymeric substances and community composition. Chlorophyll a and cyanobacterial biomass were also altered by methanol. This study suggested that the concentration-dependent effect of alachlor mainly remains limited to biomass and growth inhibition without apparent changes of structural and functional characteristics measured. Our work also establishes the potential toxic effects of organic solvents on river biofilm in ecotoxicological experiments. For the ecotoxicological experiments, the alternative of dissolution in organic solvent followed by its evaporation, depositing the chemical on a glass surface prior to dissolution in river water used here, appears to allow exposure while minimizing the effect of organic solvent.

Influence of the natural growth environment on the sensitivity of phototrophic biofilm to herbicide.

Ecotoxicological experiments were performed in laboratory-scale microcosms to investigate community-level structural responses of river phototrophic biofilms from different environments to herbicide exposure. Biofilms were initially cultivated on artificial supports placed in situ for 4 weeks at two sites, site M, located in an agricultural watershed basin and site S, located in a forested watershed basin. The biofilms were subsequently transferred to microcosms and, after an acclimatisation phase of 7 days were exposed to alachlor at 10 and 30 µg L(-1) for 23 days. Alachlor effects were assessed by a combination of structural parameters, including biomass (ash-free dry mass and chlorophyll a), molecular fingerprinting of the bacterial community (polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE)) and diatom species composition. Alachlor impacted the chlorophyll a and ash-free dry mass levels of phototrophic biofilms previously cultivated at site S. The structural responses of bacterial and diatom communities were difficult to distinguish from changes linked to the microcosm incubation period. Phototrophic biofilms from site S exposed at 30 µg L(-1) alachlor were characterised by an increase of Achnanthidium minutissimum (K-z.) Czarnecki abundance, as well as a higher proportion of abnormal frustules. Thus, phototrophic biofilms with different histories, exhibited different responses to alachlor exposure demonstrating the importance of growth environment. These observations also confirm the problem of distinguishing changes induced by the stress of pesticide toxicity from temporal evolution of the community in the microcosm.

Changes in tolerance to herbicide toxicity throughout development stages of phototrophic biofilms.

Ecotoxicological experiments have been performed in laboratory-scale microcosms to investigate the sensitivity of phototrophic biofilm communities to the alachlor herbicide, in relation to the stages of phototrophic biofilm maturation (age of the phototrophic biofilms) and physical structure (intact biofilm versus recolonization). The phototrophic biofilms were initially cultivated on artificial supports in a prototype rotating annular bioreactor (RAB) with Taylor-Couette type flow under constant operating conditions. Biofilms were collected after 1.6 and 4.4 weeks of culture providing biofilms with different maturation levels, and then exposed to nominal initial alachlor concentration of 10 µg L(-1) in either intact or recolonized biofilms for 15 days in microcosms (mean time-weighted average concentration - TWAC of 5.52 ± 0.74 µg L(-1)). At the end of the exposure period, alachlor effects were monitored by a combination of biomass descriptors (ash-free dry mass - AFDM, chlorophyll a), structural molecular fingerprinting (T-RFLP), carbon utilization spectra (Biolog) and diatom species composition. We found significant effects that in terms of AFDM, alachlor inhibited growth of the intact phototrophic biofilms. No effect of alachlor was observed on diatom composition or functional and structural properties of the bacterial community regardless of whether they were intact or recolonized. The intact three-dimensional structure of the biofilm did not appear to confer protection from the effects of alachlor. Bacterial community structure and biomass level of 4.4 weeks - intact phototrophic biofilms were significantly influenced by the biofilm maturation processes rather than alachlor exposure. The diatom communities which were largely composed of mobile and colonizer life-form populations were not affected by alachlor. This study showed that the effect of alachlor (at initial concentration of 10 µg L(-1) or mean TWAC of 5.52 ± 0.74 µg L(-1)) is mainly limited to biomass reduction without apparent changes in the ecological succession trajectories of bacterial and diatom communities and suggested that carbon utilization spectra of the biofilm are not damaged resulting. These results confirmed the importance of considering the influence of maturation processes or community age when investigating herbicide effects. This is particularly important with regard to the use of phototrophic biofilms as bio-indicators.

A photosynthetic rotating annular bioreactor (Taylor-Couette type flow) for phototrophic biofilm cultures.

In their natural environment, the structure and functioning of microbial communities from river phototrophic biofilms are driven by biotic and abiotic factors. An understanding of the mechanisms that mediate the community structure, its dynamics and the biological succession processes during phototrophic biofilm development can be gained using laboratory-scale systems operating with controlled parameters. For this purpose, we present the design and description of a new prototype of a rotating annular bioreactor (RAB) (Taylor-Couette type flow, liquid working volume of 5.04 L) specifically adapted for the cultivation and investigation of phototrophic biofilms. The innovation lies in the presence of a modular source of light inside of the system, with the biofilm colonization and development taking place on the stationary outer cylinder (onto 32 removable polyethylene plates). The biofilm cultures were investigated under controlled turbulent flowing conditions and nutrients were provided using a synthetic medium (tap water supplemented with nitrate, phosphate and silica) to favour the biofilm growth. The hydrodynamic features of the water flow were characterized using a tracer method, showing behaviour corresponding to a completely mixed reactor. Shear stress forces on the surface of plates were also quantified by computer simulations and correlated with the rotational speed of the inner cylinder. Two phototrophic biofilm development experiments were performed for periods of 6.7 and 7 weeks with different inoculation procedures and illumination intensities. For both experiments, biofilm biomasses exhibited linear growth kinetics and produced 4.2 and 2.4 mg cm(-)² of ash-free dry matter. Algal and bacterial community structures were assessed by microscopy and T-RFLP, respectively, and the two experiments were different but revealed similar temporal dynamics. Our study confirmed the performance and multipurpose nature of such an innovative photosynthetic bioreactor for phototrophic biofilm investigations.

Activated sludge as inoculum for ready biodegradability testing: effect of source.

Results of ready biodegradability tests (RBT) are barely reproducible owing to a well-known lack of definition in inoculum source and quality. In this study, the degree of variability expected when only activated sludges are used as inoculum source was investigated. For this, the characteristics of activated sludges collected in municipal wastewater treatment plants operating at various massic loading rates (MLR; 0.1, 0.5 and 0.9 kgBOD5 kgVSS(-1) d(-1)) were compared. In order to provide suitable cellular densities for RBT, inocula were obtained after settling of activated sludges and analyzed in terms of active and cultivable cell densities, dehydrogenasic activity, BOD5 and a general profile of hydrolytic enzymes. In our analysis, biomass obtained from the High-MLR treatment plant constituted the inoculum having the highest biodegradation potential both with respect to microbial densities and to enzyme activities. This biomass also yielded the fastest biodegradation kinetics in dodecyl benzene sulfonate RBT. An attempt of biomass homogenization of inocula on the basis of cultivable cell density and dehydrogenasic activity gave negative results with this chemical compound. Since, in practice, restriction of activated sludge sources may be difficult, our results emphasize the importance of further studies aimed at homogenization of inoculum quality and quantity.

Kinetics of soluble microbial product formation in substrate-sufficient batch culture of activated sludge.

The kinetics of soluble microbial product (SMP) formation under substrate-sufficient conditions appear to exhibit different patterns from substrate-limited cultures. However, energy spilling-associated SMP formation is not taken into account in the existing kinetic models and classification of SMP. Based on the concepts of growth yield and energy uncoupling, a kinetic model describing energy spilling-associated SMP formation in relation to the ratio of initial substrate concentration to initial biomass concentration ( S(0)/ X(0)) was developed for substrate-sufficient batch culture of activated sludge, and was verified by experimental data. The specific rate of energy spilling-associated SMP formation showed an increasing trend with the S(0)/ X(0) ratio up to its maximum value. The SMP productivity coefficient ( alpha(p/e)) was defined from the model on the basis of energy spilling-associated substrate consumption. Results revealed that less than 5% of energy spilling-associated substrate consumption was converted into SMP.

Excess sludge reduction in activated sludge processes by integrating biomass alkaline heat treatment.

With new EC regulations, alternative treatment and disposal techniques of the excess sludge produced by activated sludge wastewater treatment plants have to be developed. To decrease activated sludge production yield, microbial cell lysis can be amplified to enhance cryptic growth (biomass growth on lysates). Cell breakage techniques (thermal, alkaline and a combination) were studied to generate Ralstonia eutropha (strain model) and waste activated sludge lysates and to evaluate their biodegradability. Gentle treatment conditions by alkaline waste treatment (20 min at 60 degrees C and pH 10 by NaOH addition) allowed waste activated sludge to be solubilized by a two step process (instantaneous and post-treatment) giving a dissolved organic carbon released by the total suspended solids treated of 267 mgDOC x g(-1)TSS. The biodegradation of the soluble fraction of the lysates by fresh sludge reached 75 and 90% after 48 and 350 hrs of incubation respectively. A validation on a laboratory scale by insertion of a liquor alkaline heat treatment loop in a biological synthetic wastewater treatment process was carried out. A reduction of 37% of the excess sludge was obtained without altering the purification yield of the process.

Reductive cleavage of demeton-S-methyl by Corynebacterium glutamicum in cometabolism on more readily metabolizable substrates.

Corynebacterium glutamicum is able to biotransform demeton-S-methyl, an organophosphorus compound, during cometabolism with more readily metabolizable substrates. Among the cosubstrates used, fructose is the growth substrate that is most favorable for demeton-S-methyl biotransformation. The reaction mechanism of demeton-S-methyl biotransformation involves reductive cleavage of an S-C bond, which leads to accumulation of dimethyl thiophosphate in the culture medium.

Growth rate influences reductive biodegradation of the organophosphorus pesticide demeton by Corynebacterium glutamicum.

The organophosphorous pesticide, demeton-S-methyl was transformed by Corynebacterium glutamicum in co-metabolism with more readily degradable substrates. Glucose, acetate and fructose were tested as growth substrates, and the highest demeton-S-methyl biotransformation average rate (0.78 mg l(-1) h(-1)) and maximum instantaneous rate (1.4 mg l(-1) h(-1)) were achieved on fructose. This higher efficiency seems to be linked to the atypical behavior of C. glutamicum grown on fructose, characterized by a prolonged period of accelerating growth instead of a constant growth rate observed on glucose or acetate. More precisely, for growth rates in the 0.1-0.4 h(-1) range, a direct coupling between the specific demeton-S-methyl consumption rate and the growth rate was demonstrated on fructose during batch-, steady state continuous- or continuous cultures with a controlled transient growth rate (accelerostat technology). The demeton-S-methyl biotransformation was more favoured during an acceleration phase of the growth rate.

A kinetic model incorporating energy spilling for substrate removal in substrate-sufficient batch culture of activated sludge.

Batch assays are currently used to study the kinetic behavior of microbial growth. However, it has been shown that the outcome of batch experiments is greatly influenced by the initial ratio of substrate concentration (S(o)) to biomass concentration (Xo). Substrate-sufficient batch culture is known to have mechanisms of spilling energy that lead to significant nongrowth-associated substrate consumption, and the Monod equation is no longer appropriate. By incorporating substrate consumption associated with energy spilling into the balance of the substrate oxidation reaction, a kinetic model for the observed specific substrate consumption rate was developed for substrate-sufficient batch culture of activated sludge, and was further verified by experimental data. It was demonstrated that the specific substrate consumption rate increased with the increase of the S(o)/Xo ratio, and the majority of substrate was consumed through energy spilling at high S(o)/Xo ratios. It appears that the S(o)/Xo ratio is a key parameter in regulating metabolic pathways of microorganisms.

Towards a reduction in excess sludge production in activated sludge processes: biomass physicochemical treatment and biodegradation.

To decrease activated sludge production, microbial cell lysis can be amplified to enhance cryptic growth (biomass growth on lysates). Cell breakage techniques (thermal, alkaline, acid) were studied to generate Alcaligenes eutrophus and sludge lysates and to evaluate their biodegradability. Gentle treatment conditions produced the best results. Complete cell deactivation was obtained for temperatures higher than 55 degrees C. The release kinetics were similar for temperatures varying from 60 degrees C to 100 degrees C. A 20-min incubation was suitable for reaching 80% of the maximum releasable carbon. In thermal-chemical hydrolysis, NaOH was the most efficient for inducing cell lysis. Carbon release was a two-step process. First an immediate release occurred, which was of the same order of magnitude for A. eutrophus and sludge [100-200 mg dissolved organic C (DOC) g total suspended solids (TSS)-1], followed by a post-treatment release. The second step was virtually equivalent to the first for sludge, and weaker for A. eutrophus (< 50 mg DOC g TSS-1). The biodegradability of the soluble fraction, both the immediate and the post-treatment carbon release, was investigated. The optimal degradation yield, obtained with sludge cells, reached 55% after 48 h of incubation and 80% after 350 h. The most consistent lysis and biodegradation results occurred at pH 10 and 60 degrees C after a 20-min incubation.

Mechanism of enhanced oxygen transfer in fermentation using emulsified oxygen-vectors.

Limitations of oxygen transfer in fermentation can be solved using auxiliary liquids immiscible in the aqueous phase. The liquids (called oxygen-vectors) used in this study were hydrocarbon (n-dodecane) and perfluorocarbon (forane F66E) in which oxygen is highly soluble (54.9 mg/L in n-dodecane and 118 mg/L in forane F66E at 35 degrees C in contact with air at atmospheric pressure). It has been demonstrated that the use of n-dodecane emulsion in a culture of Aerobacter aerogenes enabled a 3. 5-fold increase of the volumetric oxygen transfer coefficient(k(L)a) calculated on a per-liter aqueous phase basis. The droplet size of the vector played a crucial role in the phenomena. When a static contact between gas bubble and vector droplet was established in water, the vector covered the bubble, in agreement with positive values of the spreading coefficient for these fluids. The determination of the oxygen transfer coefficients (k(L)) in a reactor with a definite interfacial area enabled the main resistance to be located in the boundary layer of the waterside either for a gas-water or a vector-water interface. Because oxygen consumption by weakly hydrophobic cells can only occur in the aqueous phase, the oxygen transfer is achieved according to the following pathway: gas-vector-water-cell. Finally, a mechanism for oxygen transfer within this four-phased system is proposed.

Enhancement of oxygen transfer rates in fermentation using oxygen-vectors.

Oxygen transfer is one of the bottlenecks in conventional fermentation technology and it has so far been almost totally overlooked with regards to high cell densities and immobilized cells. This review presents some new concepts to improve oxygen supply in aerobic fermentations, especially the use of oxygen-vectors. The oxygen-vectors generally used are liquids which are insoluble in the fermentation media. Their utilization in an emulsified form can significantly increase the oxygen transfer coefficient between gas and aqueous phases. It seems that the vector acts as an active intermediate in the oxygen transport from gas bubbles to aqueous phase, but the mechanisms involved in this unconventional technique of aeration are not yet known.