Standardization of activated sludge for biodegradation tests

Activated sludges are an inoculum source commonly used in biodegradation studies, as wastewater treatment facilities constitute an entry point to the environment for many chemicals. In this paper, the main issues relating to the use of activated sludge in biodegradability tests are presented. Special attention is also devoted to discussing the factors affecting both the activity of the microbial communities and the test results. After a short survey of the state of the art of microbiology of activated sludge, the paper focuses on the methods used to reduce the variations in the diversity, quality and quantity of these communities. Finally, use of surrogates as reference materials in biodegradability tests is discussed

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

Ecotoxicological experiments have been performed in laboratory-scale microcosms to investigate thesensitivity of phototrophic biofilm communities to the alachlor herbicide, in relation to the stages ofphototrophic biofilm maturation (age of the phototrophic biofilms) and physical structure (intact biofilmversus recolonization). The phototrophic biofilms were initially cultivated on artificial supports in aprototype rotating annular bioreactor (RAB) with Taylor–Couette type flow under constant operatingconditions. Biofilms were collected after 1.6 and 4.4 weeks of culture providing biofilms with differentmaturation levels, and then exposed to nominal initial alachlor concentration of 10 ug L−1in either intactor recolonized biofilms for 15 days in microcosms (mean time-weighted average concentration – TWACof 5.52 ± 0.74 ug L−1).At the end of the exposure period, alachlor effects were monitored by a combination of biomass descrip-tors (ash-free dry mass – AFDM, chlorophyll a), structural molecular fingerprinting (T-RFLP), carbonutilization spectra (Biolog) and diatom species composition. We found significant effects that in terms ofAFDM, alachlor inhibited growth of the intact phototrophic biofilms. No effect of alachlor was observedon diatom composition or functional and structural properties of the bacterial community regardless ofwhether they were intact or recolonized. The intact three-dimensional structure of the biofilm did notappear to confer protection from the effects of alachlor. Bacterial community structure and biomass levelof 4.4 weeks – intact phototrophic biofilms were significantly influenced by the biofilm maturation pro-cesses rather than alachlor exposure. The diatom communities which were largely composed of mobileand colonizer life-form populations were not affected by alachlor.This study showed that the effect of alachlor (at initial concentration of 10 ug L−1or mean TWAC of5.52 ± 0.74 ug L−1) is mainly limited to biomass reduction without apparent changes in the ecologicalsuccession trajectories of bacterial and diatom communities and suggested that carbon utilization spec-tra of the biofilm are not damaged resulting. These results confirmed the importance of consideringthe influence of maturation processes or community age when investigating herbicide effects. This isparticularly 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) (TayloreCouette 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

Mangrove Facies Drives Resistance and Resilience of Sediment Microbes Exposed to Anthropic Disturbance

Mangrove forests are coastal ecosystems continuously affected by various environmental stresses and organized along constraint gradients perpendicular to the coastline. The aim of this study was to evaluate the resistance and resilience of sediment microbial communities in contrasted vegetation facies, during and after exposure to an anthropic disturbance. Our hypothesis was that microbial communities should be the most stable in the facies where the consequences of the anthropic disturbance are the most similar to those of natural disturbances. To test this, we focused on communities involved in N-cycle. We used an in situ experimental system set up in Mayotte Island where 2 zones dominated by different mangrove trees are daily exposed since 2008 to pretreated domestic wastewater (PW) discharges. These freshwater and nutrients inputs should increase microbial activities and hence the anoxia of sediments. We monitored during 1 year the long-term impact of this disturbance, its short-term impact and the resilience of microbial communities on plots where PW discharges were interrupted. Microorganism densities were estimated by qPCR, the nitrification (NEA) and denitrification (DEA) enzyme activities were evaluated by potential activity measurements and pigment analyses were performed to assess the composition of microbial photosynthetic communities. At long-term PW discharges significantly modified the structure of phototrophic communities and increased the total density of bacteria, the density of denitrifying bacteria and DEA. Similar effects were observed at short-term, notably in the facies dominated by Ceriops tagal. The results showed a partial resilience of microbial communities. This resilience was faster in the facies dominated by Rhizophora mucronata, which is more subjected to tides and sediment anoxia. The higher stability of microbial communities in this facies confirms our hypothesis. Such information should be taken into account in mangrove utilization and conservation policies

Dispersed ice of permafrost peatlands represents an important source of labile carboxylic acids, nutrients and metals

Thawing of frozen organic and mineral soils and liberation of organic carbon (OC), macro- and micro-nutrients and trace elements from pore ice in high latitude regions represent a potentially important but poorly quantified retroactive linkage to climate warming. This is especially true for permafrost peatlands, occupying a sizable proportion of all permafrost territories and presenting a large and highly vulnerable stock of soil OC which can be subjected to fast thawing at currently circum-zero temperatures. The conventional method of assessing the labile water-soluble fraction of permafrost soils is aqueous extraction from dried soil. However, this technique does not allow collecting native ice present in soil pores and is therefore likely to underestimate or overestimate the pool of labile soil C and nutrients. Here, we present results of direct pore ice analyses performed on native peat cores from the western Siberia Lowland in comparison to the water extraction (10 and 100 gdry peat L-1) of soluble components from the same peat subjected to freeze drying. Aqueous leachates of permafrost peat from both thawed (0–45 cm) and frozen (45–130 cm) layers yielded high concentrations of DOC, nutrients, carboxylic acids and trace metals, comparable or higher to those in peat porewater and dispersed peat ice. We found strong (a factor of 3 to 30) enrichment in the frozen part of the core (below 45 cm, which is active layer depth) in dissolved OC, many carboxylates (acetate, formate, lactate, butyrate, propionate, pyruvate), inorganic nutrients (Si, P, N) and trace elements (Fe, Al, Mn, Zn, Sr and Ba). The dispersed ice which is present in peat below active layer represents highly labile reservoir of organic and inorganic nutrients which should be considered in permafrost thaw scenario

Autogenic versus environmental control during development of river biofilm

In the natural environment, microbial community structure of river biofilm is controlled by biotic and abiotic factors. This study explored the capacity to manipulate the structure of microbial communities by modifying environmental conditions during the course of biofilm development. River epilithic biofilm was cultivated in situ on artificial substrates placed parallel to river water flow. Substrates were incubated for 3 and 5.5 weeks in river to allow natural biofilm development, at two sites with contrasting physico-chemical characteristics. The first site (Aurade´ , Gers, France) was located in an agricultural watershed basin and the second site (Larroque, Haute-Garonne, France) was located in a forested watershed basin. After 3 weeks of biofilm development, a subset of substrates was collected from one site and transplanted to the second site where they remained for 2.5 further weeks. Epilithic bacterial community structure (at 3 weeks from each site and at 5.5 weeks from biofilms with and without transplantation) was assessed using PCR-DGGE of 16S rDNA fragment. Biofilm biomass was estimated using ash free dry mass (AFDM). After 3 weeks of development, biofilms from the two sites exhibited comparable AFDM values (average of 1.4¡0.2 g.mx2). A difference between the two sites was observed after 5.5 weeks of development: AFDM decreased for biofilms from the agricultural watershed basin (from 1.4 to 0.18 g.mx2) as a consequence of grazing pressure (Bithynia), and increased for biofilms from the forested agricultural watershed (from 1.4 to 2.6 g.mx2). Microbial community analyses revealed a differentiated community structure between biofilms from the different sites and exhibited a change of microbial community structure after 5.5 weeks of biofilm development. These observations confirm a process of ecological succession in microbial communities. Changing the incubation site during biofilm development modified the trajectory of these ecological successions, suggesting that site characteristics mainly conditioned the structure of these microbial communities

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

Caractérisation, dynamique et facteurs de contrôle des communautés bactériennes de biofilms de rivière

TOULOUSE3-BU Sciences (315552104) / SudocSudocFranceF

ETUDE DES POSSIBILITES DE STANDARDISATION DES INOCULA DES TESTS DE BIODEGRADABILITE FACILE (CAS DES BOUES ACTIVEES DE STATION D'EPURATION)

TOULOUSE-INSA (315552106) / SudocSudocFranceF

Carbon sequestration potential of Mg carbonate and silicate biomineralization in the presence of cyanobacterium Synechococcus

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