Lorena Lima de Oliveira

Anaerobic degradation of linear alkylbenzene sulfonate (LAS) in fluidized bed reactor by microbial consortia in different support materials.

Four anaerobic fluidized bed reactors filled with activated carbon (R1), expanded clay (R2), glass beads (R3) and sand (R4) were tested for anaerobic degradation of LAS. All reactors were inoculated with sludge from a UASB reactor treating swine wastewater and were fed with a synthetic substrate supplemented with approximately 20 mg l(-1) of LAS, on average. To 560 mg l(-1) COD influent, the maximum COD and LAS removal efficiencies were mean values of 97+/-2% and 99+/-2%, respectively, to all reactors demonstrating the potential applicability of this reactor configuration for treating LAS. The reactors were kept at 30 degrees C and operated with a hydraulic retention time (HRT) of 18h. The use of glass beads and sand appear attractive because they favor the development of biofilms capable of supporting LAS degradation. Subsequent 16S rRNA gene sequencing and phylogenetic analysis of samples from reactors R3 and R4 revealed that these reactors gave rise to broad microbial diversity, with microorganisms belonging to the phyla Bacteroidetes, Firmicutes, Actinobacteria and Proteobacteria, indicating the role of microbial consortia in degrading the surfactant LAS.

Development of a method by HPLC to determine LAS and its application in anaerobic reactors

This work describes the development and validation of a method using High Performance Liquid Chromatography (HPLC) for Linear Alkylbenzene Sulfonate (LAS) determination in sample of wastewater from anaerobic reactors. The applied LAS standard was the dodecylbenzene sulfonic acid-sodium salt, which presents four main peaks in the chromatogram, related to different homologous of the linear alkyl chain. Different chromatographic conditions were tested with C-18, C-12 and C-8 as stationary phases, ultraviolet and fluorescence detectors, mobile phase (MP) compositions and programming time of elution gradient. The best chromatographic condition was obtained with the C-8 column, MP: methanol and of sodium perchlorate (0.075 mol L-1). The validation of this method was made with the calibration of LAS curves using water and synthetic substrate as solvents. The method was validated in order to demonstrate its precision, linearity, limit of detection of each homolog and its instrumental precision.

Treatment of linear alkylbenzene sulfonate in a horizontal anaerobic immobilized biomass reactor.

Linear alkylbenzene sulfonate (LAS) is an anionic surfactant widely used to manufacture detergents and found in domestic and industrial wastewater. LAS removal was evaluated in a horizontal anaerobic immobilized biomass reactor. The system was filled with polyurethane foam and inoculated with sludge that was withdrawn from an up flow anaerobic sludge blanket reactor that is used to treat swine wastewater. The reactor was fed with easily degradable substrates and a solution of commercial LAS for 313 days. The hydraulic retention time applied was 12h. The system was initially operated without detergent and resulted to 94% reduction of demand. The mass balance in the system indicated that the LAS removal efficiency was 45% after 18 0days. From the 109 th day to the 254 th day, a removal efficiency of 32% was observed. The removal of LAS was approximately 40% when 1500 mg of LAS were applied in the absence of co-substrates suggesting that the LAS molecules were used selectively. Microscopic analyses of the biofilm revealed diverse microbial morphologies and denaturing gradient gel electrophoresis profiling showed variations in the total bacteria and sulfate-reducing bacteria populations. 16S rRNA sequencing and phylogenetic analyses demonstrated that members of the order Clostridiales were the major components of the bacterial community in the last step of the reactor operation.

Influence of support material on the immobilization of biomass for the degradation of linear alkylbenzene sulfonate in anaerobic reactors

Two horizontal-flow anaerobic immobilized biomass reactors (HAIB) were used to study the degradation of the LAS surfactant: one filled with charcoal (HAIB1) and the other with a mixed bed of expanded clay and polyurethane foam (HAIB2). The reactors were fed with synthetic substrate supplemented with 14 mg l(-1)of LAS, kept at 30+/-2 degrees C and operated with a hydraulic retention time (HRT) of 12h. The surfactant was quantified by HPLC. Spatial variation analyses were done to quantify organic matter and LAS consumption along the reactor length. The presence of the surfactant in the load did not affect the removal of organic matter (COD), which was close to 90% in both reactors for an influent COD of 550 mg l(-1). The results of a mass balance indicated that 28% of all LAS added to HAIB1 was removed by degradation. HAIB2 presented 27% degradation. Molecular biology techniques revealed microorganisms belonging the uncultured Holophaga sp., uncultured delta Proteobacterium, uncultured Verrucomicrobium sp., Bacteroides sp. and uncultured gamma Proteobacterium sp. The reactor with biomass immobilized on charcoal presented lower adsorption and a higher kinetic degradation coefficient. So, it was the most suitable support for LAS anaerobic treatment.

Las degradation in a fluidized bed reactor and phylogenetic characterization of the biofilm

A fluidized bed reactor was used to study the degradation of the surfactant linear alkylbenzene sulfonate (LAS). The reactor was inoculated with anaerobic sludge and was fed with a synthetic substrate supplemented with LAS in increasing concentrations (8.2 to 45.8 mg l-1). The removal efficiency of 93% was obtained after 270 days of operation. Subsequently, 16S rRNA gene sequencing and phylogenetic analysis of the sample at the last stage of the reactor operation recovered 105 clones belonging to the domain Bacteria. These clones represented a variety of phyla with significant homology to Bacteroidetes (40%), Proteobacteria (42%), Verrucomicrobia (4%), Acidobacteria (3%), Firmicutes (2%), and Gemmatimonadetes (1%). A small fraction of the clones (8%) was not related to any phylum. Such phyla variety indicated the role of microbial consortia in degrading the surfactant LAS.

Anaerobic degradation of linear alkylbenzene sulfonate in fluidized bed reactor

An anaerobic fluidized bed reactor was used to assess the degradation of the surfactant linear alkylbenzene sulfonate (LAS). The reactor was inoculated with sludge from an UASB reactor treating swine wastewater and was fed with a synthetic substrate supplemented with LAS. Sand was used as support material for biomass immobilization. The reactor was kept in a controlled temperature chamber (30±1 oC) and operated with a hydraulic retention time (HRT) of 18 h. The LAS concentration was gradually increased from 8.2±1.3 to 45.8±5.4 mg.L-1. The COD removal was 91%, on average, when the influent COD was 645±49 mg.L-1. The results obtained by chromatographic analysis showed that the reactor removed 93% of the LAS after 270 days of operation.

Evaluation of the microbial diversity in sequencing batch reactor treating linear alkylbenzene sulfonate under denitrifying and mesophilic conditions using swine sludge as inoculum

The objective of this study was to evaluate the degradation of Linear Alkylbenzene Sulfonate (LAS) in anaerobic sequencing batch reactor (ASBR) under denitrifying conditions using swine sludge as inoculum. The reactor was operated for 104 days with synthetic substrate containing nitrate, and LAS was added later (22 mg/L). Considering the added mass of the LAS, the adsorbed mass in the sludge and discarded along with the effluent, degradation of the surfactant at the end of operation was 87%, removal of chemical oxygen demand was 86% and nitrate was 98%. The bacterial community was evaluated by cutting the bands and sequencing of polymerase chain reaction (PCR) fragments and denaturing gradient gel electrophoresis (DGGE). The sequences obtained were related to the phylum Proteobacteria and the alpha-and beta-proteobacteria classes, these bacteria were probably involved in the degradation of LAS. The efficiently degraded LAS in the reactor was operated in batch sequences in denitrifying conditions.