Sewage contamination under water scarcity effects on stream biota: biofilm, grazers, and their interaction.

One of the most common anthropogenic impacts on river ecosystems is the effluent discharge from wastewater treatment plants. The effects of this contamination on stream biota may be intensified in Mediterranean climate regions, which comprise a drought period that leads to flow reduction, and ultimately to stagnant pools. To assess individual and combined effects of flow stagnation and sewage contamination, biofilm and gastropod grazers were used in a 5-week experiment with artificial channels to test two flow velocity treatments (stagnant flow/basal flow) and two levels of organic contamination using artificial sewage (no sewage input/sewage input). Stressors' effects were determined on biofilm total biomass and chlorophyll (Chl) content, on oxygen consumption and growth rate of the grazers (Theodoxus fluviatilis), and on the interaction grazer-biofilm given by grazer's feeding activity (i.e., biofilm consumption rate). The single effect of sewage induced an increase in biofilm biomass and Chl-a content, simultaneously increasing both grazers' oxygen consumption and their feeding activity. Diatoms showed a higher sensitivity to flow stagnation, resulting in a lower content of Chl-c. Combined stressors interacted antagonistically for biofilm total biomass, Chl-b contents, and grazers's feeding rate. The effect of sewage increasing biofilm biomass and grazing activity was reduced by the presence of flow stagnation (antagonist factor). Our findings suggest that sewage contamination has a direct effect on the functional response of primary producers and an indirect effect on primary consumers, and this effect is influenced by water flow stagnation.

Application of microbial fuel cell technology for vinasse treatment and bioelectricity generation.

OBJECTIVE: Our study evaluated the performance of different two-chambered microbial fuel cell (MFC) prototypes, operated with variable distance between electrodes and Nafion membrane and specific inoculum concentration, applied for vinasse treatment. RESULTS: The performance of the developed MFC resulted in a maximum current density of 1200 mA m-2 and power density of 800 mW m-2 in a period of 61 days. MFC performed a chemical oxygen demand removal at a rate ranging from 51 to 60%. CONCLUSIONS: Taking our preliminary results into consideration, we concluded that the MFC technology presents itself as highly promising for the treatment of vinasse.

Macroinvertebrate short-term responses to flow variation and oxygen depletion: A mesocosm approach.

In Mediterranean rivers, water scarcity is a key stressor with direct and indirect effects on other stressors, such as water quality decline and inherent oxygen depletion associated with pollutants inputs. Yet, predicting the responses of macroinvertebrates to these stressors combination is quite challenging due to the reduced available information, especially if biotic and abiotic seasonal variations are taken under consideration. This study focused on the response of macroinvertebrates by drift to single and combined effects of water scarcity and dissolved oxygen (DO) depletion over two seasons (winter and spring). A factorial design of two flow velocity levels - regular and low (vL) - with three levels of oxygen depletion - normoxia, medium depletion (dM) and higher depletion (dH) - was carried out in a 5-artificial channels system, in short-term experiments. Results showed that both stressors individually and together had a significant effect on macroinvertebrate drift ratio for both seasons. Single stressor effects showed that macroinvertebrate drift decreased with flow velocity reduction and increased with DO depletion, in both winter and spring experiments. Despite single stressors opposing effects in drift ratio, combined stressors interaction (vL×dM and vL×dH) induced a positive synergistic drift effect for both seasons, but only in winter the drift ratio was different between the levels of DO depletion. Stressors interaction in winter seemed to intensify drift response when reached lower oxygen saturation. Also, drift patterns were different between seasons for all treatments, which may depend on individual's life stage and seasonal behaviour. Water scarcity seems to exacerbate the oxygen depletion conditions resulting into a greater drifting of invertebrates. The potential effects of oxygen depletion should be evaluated when addressing the impacts of water scarcity on river ecosystems, since flow reductions will likely contribute to a higher oxygen deficit, particularly in Mediterranean rivers.

Removal of tetracycline from contaminated water by Moringa oleifera seed preparations.

The aim of this study was to evaluate tetracycline antibiotic (TA) removal from contaminated water by Moringa oleifera seed preparations. The composition of synthetic water approximate river natural contaminated water and TA simulated its presence as an emerging pollutant. Interactions between TA and protein preparations (extract; fraction and lectin) were also evaluated. TA was determined by solid-phase extraction followed by high-performance liquid chromatography-mass spectrometry. Moringa extract and flour removed TA from water. The extract removed TA in all concentrations, and better removal (40%) was obtained with 40 mg L(-1); seed flour (particles < 5 mm), 1.25 and 2.50 g L(-1) removed 28% and 29% of tetracycline, respectively; particles > 5 mm (0.50 g L(-1)) removed 55% of antibiotic. Interactions between TA and seed preparations were assayed by haemagglutinating activity (HA). Specific HA (SHA) of extract (pH 7) was abolished with tetracycline (5 mg L(-1)); fraction (75%) and lectin HA (97%) were inhibited with TA. Extract SHA decreased by 75% at pH 8. Zeta potential (ZP) of extract 700 mg L(-1) and tetracycline 50 mg L(-1) , pH range 5-8, showed different results. Extract ZP was more negative (-10.73 to -16.00 mV) than tetracycline ZP (-0.27 to -20.15 mV); ZP difference was greater in pH 8. The focus of this study was achieved since Moringa preparations removed TA from water and compounds interacting with tetracycline involved at least lectin-binding sites. This is a natural process, which do not promote environmental damage.

Influence of tetracycline on the microbial community composition and activity of nitrifying biofilms.

The present work aims to evaluate the bacterial composition and activity (carbon and nitrogen removal) of nitrifying biofilms exposed to 50 µg L(-1) of tetracycline. The tetracycline removal efficiency and the occurrence of tetracycline resistance (tet) genes were also studied. Two sequencing batch biofilm reactors (SBBRs) fed with synthetic wastewater were operated without (SBBR1) and with (SBBR2) the antibiotic. Both SBBRs showed similar organic matter biodegradation and nitrification activity. Tetracycline removal was about 28% and biodegradation was probably the principal removal mechanism of the antibiotic. Polymerase chain reaction-denaturing gradient gel electrophoresis analysis of the bacterial community showed shifts leading to not only the fading of some ribotypes, but also the emergence of new ones in the biofilm with tetracycline. The study of the tet genes showed that tet(S) was only detected in the biofilm with tetracycline, suggesting a relationship between its occurrence and the presence of the antibiotic.

A flat microbial fuel cell for decentralized wastewater valorization: process performance and optimization potential.

A very compact flat microbial fuel cell (MFC), with 64 cm2 each for the anode surface and the cathode surface and 1 cm3 each for the anode and cathode chambers, was tested for wastewater treatment with simultaneous electricity production with the ultimate goal of implementing an autonomous service in decentralized wastewater treatment systems. The MFC was operated with municipal wastewater in sequencing batch reactor mode with re-circulation. Current densities up to 407 W/m3 and a carbon removal of 83% were obtained. Interruption in the operation slightly decreased power density, while the re-circulation ratio did not influence power generation. The anode biofilm presented high conductivity, activity and diversity. The denaturing gradient gel electrophoresis band-pattern of the DNA showed the presence of several ribotypes with different species of Shewanellaceae and Geobacteraceae families.

Coagulant properties of Moringa oleifera protein preparations: application to humic acid removal.

This work aimed to characterize the coagulant properties of protein preparations from Moringa oleifera seeds in the removal of humic acids from water. Three distinct preparations were assayed, namely extract (seeds homogenized with 0.15 M NaCl), fraction (extract precipitated with 60% w/v ammonium sulphate) and cMoL (protein purified with guar gel column chromatography). The extract showed the highest coagulant activity in a protein concentration between 1 mg/L and 180 mg/L at pH 7.0. The zeta potential of the extract (-10 mV to -15 mV) was less negative than that of the humic acid (-41 mV to -42 mV) in a pH range between 5.0 and 8.0; thus, the mechanism that might be involved in this coagulation activity is adsorption and neutralization of charges. Reduction of total organic carbon (TOC) and dissolved organic carbon (DOC) was observed in water samples containing 9 mg/L carbon as humic acid when treated with 1 mg/L of the extract. A decrease in colour and in the aromatic content of the treated water was also observed. These results suggested that the extract from M. oleifera seeds in a low concentration (1 mg/L) can be an interesting natural alternative for removing humic acid from water in developing countries. The extract dose determined in the present study does not impart odour or colour to the treated water.

Influence of the organic loading rate on the growth of Galactomyces geotrichum in activated sludge.

Recent studies reporting on the useful implementation of fungi in wastewater treatment plants triggered the need to improve fungi based systems. Therefore, it is crucial to investigate the conditions that promote their selection. The present work aims to study the effect of the organic loading rate on the growth of filamentous fungi. Three sequencing batch reactors (SBR), fed with an easily biodegradable substrate (acetate), were operated at different organic loading rates: 4.3 g COD L(-1) day(-1) (SBR1), 1.0 g COD L(-1) day(-1) (SBR2) and 0.5 g COD L(-1) day(-1) (SBR3). High amounts of fungal filaments were observed in the SBR operating at higher organic loading rate, as ascertained by direct microscopic inspection, while, at lower organic loading rates, overabundance of fungal filaments was not observed. Sequence retrieved from the isolated fungal filaments presented high similarity (99 %) to Galactomyces geotrichum.

Structure and activity of lacustrine sediment bacteria involved in nutrient and iron cycles.

Knowledge of the bacterial community structure in sediments is essential to better design restoration strategies for eutrophied lakes. In this regard, the aim of this study was to quantify the abundance and activity of bacteria involved in nutrient and iron cycling in sediments from four Azorean lakes with distinct trophic states (Verde, Azul, Furnas and Fogo). Inferred from quantitative PCR, bacteria performing anaerobic ammonia oxidation were the most abundant in the eutrophic lakes Verde, Azul and Furnas (4.5-16.6%), followed by nitrifying bacteria (0.8-13.0%), denitrifying bacteria (DNB) (0.5-6.8%), iron-reducing bacteria (0.2-1.4%) and phosphorus-accumulating organisms (<0.3%). In contrast, DNB dominated sediments from the oligo-mesotrophic lake Fogo (8.8%). Activity assays suggested that bacteria performing ammonia oxidation (aerobic and anaerobic), nitrite oxidation, heterothrophic nitrate reduction, iron reduction and biological phosphorus storage/release were present and active in all Azorean lake sediments. The present work also suggested that the activity of DNB might contribute to the release of phosphorus from sediments.

In situ microbial fuel cell-based biosensor for organic carbon.

The biological oxygen demand (BOD) may be the most used test to assess the amount of pollutant organic matter in water; however, it is time and labor consuming, and is done ex-situ. A BOD biosensor based on the microbial fuel cell principle was tested for online and in situ monitoring of biodegradable organic content of domestic wastewater. A stable current density of 282±23mA/m(2) was obtained with domestic wastewater containing a BOD(5) of 317±15mg O(2)/L at 22±2°C, 1.53±0.04mS/cm and pH 6.9±0.1. The current density showed a linear relationship with BOD(5) concentration ranging from 17±0.5mg O(2)/L to 78±7.6mg O(2)/L. The current generation from the BOD biosensor was dependent on the measurement conditions such as temperature, conductivity, and pH. Thus, a correction factor should be applied to measurements done under different environmental conditions from the ones used in the calibration. These results provide useful information for the development of a biosensor for real-time in situ monitoring of wastewater quality.

Towards implementation of a benthic microbial fuel cell in lake Furnas (Azores): phylogenetic affiliation and electrochemical activity of sediment bacteria.

This work was conducted to examine the composition and electrochemical activity of the bacterial community inhabiting lake Furnas sediments (Azores). Fingerprinting analysis of the bacterial 16S rRNA gene fragment was done by denaturing gradient gel electrophoresis. The sequences retrieved from lake Furnas sediments were affiliated to Bacteroidetes/Chlorobi group, Chloroflexi, Alfa-, Delta-, and Gamma-subclasses of Proteobacteria, Cyanobacteria, and Gemmatimonadetes. A cyclic voltammetric study was carried out with an enriched sediment bacterial suspension in a standard two chamber electrochemical cell using a carbon paper anode. Cyclic voltammograms (scan rate of 50 mV/s) showed the occurrence of oxidation-reduction reactions at the carbon anode surface. The benthic microbial fuel cell operated with lake Furnas sediments presented a low power density (1 mW/m(2)) indicating that further work is required to optimise its power generation. These results suggested that sediment bacteria, probably from the Delta- and Gamma-subclasses of Proteobacteria, were electroactive under tested conditions.

Determination of total and available fractions of PAHs by SPME in oily wastewaters: overcoming interference from NAPL and NOM.

BACKGROUND, AIM, AND SCOPE: Polycyclic aromatic hydrocarbons (PAHs) are often found in oily wastewaters. Their presence is usually the result of human activities and has a negative effect on the environment. One important step in addressing this problem is to evaluate the effectiveness of PAH removal by biological processes since these are the most cost-effective treatments known today. Many techniques are presently available for PAH determination in wastewaters. Solid phase microextracion (SPME) is known to be one of the most effective techniques for this purpose. When analyzing complex matrices with substances such as natural organic matter (NOM) and non-aqueous phase liquids (NAPL), it is important to differentiate the free dissolved PAH from matrix-bonded PAH. PAHs associated with the bonded fraction are less susceptible to biological treatment. The present study concerns the development of a simple and suitable methodology for the determination of the freely dissolved and the total fraction of PAHs present in oily wastewaters. The methodology was then applied to an oily wastewater from a fuel station retention basin. MATERIAL AND METHODS: Headspace SPME was used for analyzing PAH since the presence of a complex or dirty matrix in direct contact with the fiber may damage it. Four model PAHs-anthracene, fluorene, phenanthrene, and pyrene-were analyzed by GC-MS. Negligible depletion SPME technique was used to determine the free fraction. Total PAH was determined by enhancing the mass transfer from the bonded phase to the freely dissolved phase by temperature optimization and the use of the method of standard additions. The PAH absorption kinetics were determined in order to define the optimal sampling conditions for this method. The fitting of the experimental data to a mathematical model was accomplished using Berkeley Madonna software. Humic acid and silicon oil were used as model NOM and NAPL, respectively, to study the effect of these compounds on the decrease of SPME response. Then, the method was evaluated with wastewater from a fuel station spill retention basin. RESULTS: The SPME kinetic parameters-k (1) (uptake rate), k (2) (desorption rate), and K (SPME) (partition coefficient)-were determined from experimental data modeling. The determination of the free fraction required 15-min sampling to ensure that PAH depletion from sample was below 1%. For total PAH, a 30-min extraction at 100 degrees C ensured the maximum signal response in the GC-MS. For the determination of free and total PAHs, extractions were performed before reaching the SPME equilibrium. The wastewater used in this study had no free fraction of the analyzed PAHs. However, the four studied PAHs were found when the method for total PAH was used. DISCUSSION: The addition of NOM and NAPL dramatically decreased the efficiency of the SPME. This decrease was the result of a greater partition of the PAHs to the NAPL and NOM phases. This fact was also observed in the analysis of the fuel station spill retention basin, where no free PAH was measured. However, using the method of standard addition for the determination of total PAH, it was possible to quantify all four PAHs. CONCLUSIONS: The method developed in the present study was found to be adequate to differentiate between free and total PAH present in oily wastewater. It was determined that the presence of NOM and NAPL had a negative effect on SPME efficiency. RECOMMENDATIONS AND PERSPECTIVES: The presence of binding substances had a great influence on SPME kinetics. Therefore, it is of extreme importance to determine their degree of interference when analyzing oily wastewaters or results can otherwise be erroneous. Other factors influencing the total PAH determinations should be considered in further studies.

Synthesis and degradation of poly-beta-hydroxybutyrate in a sequencing batch biofilm reactor.

The aim of this work was the study of poly-beta-hydroxybutyrate (PHB) formation and degradation in a sequencing batch biofilm reactor (SBBR). The SBBR was operated in cycles comprising three individual phases: mixed fill, aeration and draw. A synthetic substrate solution with acetate and ammonium was used. PHB was formed during the aeration phase immediately after acetate depletion, and was subsequently consumed for biomass growth, owing to the high oxygen concentration in the reactor. It was observed a combination of suspended and biofilm growth in the SBBR with predominance of the fixed form of biomass (506 Cmmol and 2102 Cmmol, respectively). Maximum PHB fraction of suspended biomass (0.13 Cmol/Cmol) was considerably higher than that of biofilm (0.01 Cmol/Cmol). This may possibly be explained by a combination of two factors: lower mass transfer limitation of acetate and higher fraction of heterotrophs in suspended biomass compared to the ones of biofilm.

Fluorene and phenanthrene uptake by Pseudomonas putida ATCC 17514: kinetics and physiological aspects.

Pseudomonas putida ATCC 17514 was used as a model strain to investigate the characteristics of bacterial growth in the presence of solid fluorene and phenanthrene. Despite the lower water-solubility of phenanthrene, P. putida degraded this polycyclic aromatic hydrocarbon (PAH) at a maximum observed rate of 1.4 +/- 0.1 mg L(-1) h(-1), higher than the apparent degradation rate of fluorene, 0.8 +/- 0.07 mg L(-1) h(-1). The role of physiological processes on the biodegradation of these PAHs was analyzed and two different uptake strategies were identified. Zeta potential measurements revealed that phenanthrene-grown cells were slightly more negatively charged (-57.5 +/- 4.7 mV) than fluorene-grown cells (-51.6 +/- 4.9 mV), but much more negatively charged than glucose-grown cells (-26.8 +/- 3.3 mV), suggesting that the PAH substrate induced modifications on the physical properties of bacterial surfaces. Furthermore, protein-to-exopolysaccharide ratios detected during bacterial growth on phenanthrene were typical of biofilms developed under physicochemical stress conditions, caused by the presence of sparingly water-soluble chemicals as the sole carbon and energy source for growth, the maximum value for TP/EPS during growth on phenanthrene (1.9) being lower than the one obtained with fluorene (5.5). Finally, confocal laser microscopy observations using a gfp-labeled derivative strain revealed that, in the presence of phenanthrene, P. putida::gfp cells formed a biofilm on accessible crystal surfaces, whereas in the presence of fluorene the strain grew randomly between the crystal clusters. The results showed that P. putida was able to overcome the lower aqueous solubility of phenanthrene by adhering to the solid PAH throughout the production of extracellular polymeric substances, thus promoting the availability and uptake of such a hydrophobic compound.