Microbial interactions and nitrogen removal performance in an intermittently rotating biological contactor treating mature landfill leachate.

Developing efficient landfill leachate treatment is still necessary to reduce environmental risks. However, nitrogen removal in biological treatment systems is often poor or costly. Studying biofilms in anoxic/aerobic zones of rotating biological contactors (RBC) can elucidate how microbial interactions confer resistance to shock loads and toxic substances in leachate treatment. This study assessed the nitritation-anammox performance in an intermittent-rotating bench-scale RBC treating mature leachate (diluted). Despite the leachate toxicity, the system achieved nitritation with an efficiency of up to 34 % under DO values between 0.8 and 1.8 mg.L-1. The highest average ammoniacal nitrogen removal was 45.3 % with 10 h of HRT. The 16S rRNA sequencing confirmed the presence of Nitrosonomas, Aquamicrobium, Gemmata, and Plantomyces. The coexistence of these bacteria corroborated the selective pressure exerted by leachate in the community structure. The microbial interactions found here highlight the potential application of RBC to remove nitrogen in landfill leachate treatment.

Biofilm stratification and autotrophic-heterotrophic interactions in a structured bed reactor (SBRIA) for carbon and nitrogen removal.

The structured-bed reactor with intermittent aeration (SBRIA) is a promising technology for simultaneous carbon and nitrogen removal from wastewater. An in depth understanding of the microbiological in the reactor is crucial for its optimization. In this research, biofilm samples from the aerobic and anoxic zones of an SBRIA were analyzed through 16S rRNA sequencing to evaluate the bacterial community shift with variations in the airflow and aeration time. The control of the airflow and aeration time were essential to guarantee reactor performances to nitrogen removal close to 80%, as it interfered in nitrifying and denitrifying communities. The aeration time of 1.75 h led to establishment of different nitrogen removal pathways by syntrophic relationships between nitrifier, denitrifier and anammox species. Additionally, the predominance of these different species in the internal and external parts of the biofilm varied according to the airflow.

Optimization of airflow and aeration cycles in a new structured bed reactor configuration for carbon and nitrogen removal.

The Structured Bed Reactor with Recirculation and Intermittent Aeration (SBRRIA) is a reactor configuration that presents high efficiency of organic matter and nitrogen removal, besides low sludge production. However, operational parameters, as the recirculation rate, aeration time, and airflow, are not fully established. A bench-scale structured bed reactor with intermittent aeration was fed with synthetic effluent simulating the characteristics of sanitary sewage. The reactor was operated for 280 days with an operational hydraulic retention time (HRT) of 10 h. The reactor was operated without effluent recirculation for the first time since this approach was not yet reported, and was named Structured Bed Reactor with Intermittent Aeration (SBRIA). The COD removal was higher than 81% for all operational conditions, and the total nitrogen removal ranged from 10 to 80%. The highest efficiencies were obtained with an aeration time of 1 h 45 min (total cycle of 3 h) and an airflow rate of 4.5 L.min-1. Different nitrification and denitrification behaviours were observed, resulting in nitrification efficiencies over 90% when the reactor was submitted to higher aeration times and denitrification efficiencies above 90% when the reactor was submitted to low aeration times. The airflow ranges tested in this study affected the nitrification and the total nitrogen efficiencies. Even without effluent recirculation, the temporal profile showed that there were no peaks in the concentration of the nitrogen forms in the reactor effluent, saving electrical energy up to 75% due to pumping.

Microbiome taxonomic and functional profiles of two domestic sewage treatment systems.

Anaerobic systems for domestic sewage treatment, like septic tanks and anaerobic filters, are used in developing countries due to favorable economic and functional features. The anaerobic filter is used for the treatment of the septic tank effluent, to improve the COD removal efficiency of the system. The microbial composition and diversity of the microbiome from two wastewater treatment systems (factory and rural school) were compared through 16S rRNA gene sequencing using MiSeq 2 × 250 bp Illumina sequencing platform. Additionally, 16S rRNA data were used to predict the functional profile of the microbial communities using PICRUSt2. Results indicated that hydrogenotrophic methanogens, like Methanobacterium, were found in higher abundance in both systems compared to acetotrophic methanogens belonging to Methanosaeta genus. Also, important syntrophic microorganisms (Smithella, Syntrophus, Syntrophobacter) were found in the factory and rural school wastewater treatment systems. Microbial communities were also compared between stages (septic tank and anaerobic filter) of each wastewater treatment stage, revealing that, in the case of the rural school, both microbial communities were quite similar most likely due to hydraulic short-circuit issues. Meanwhile, in the factory, microbial communities from the septic tank and anaerobic filter were different. The school system showed lower COD removal rates (2-30%), which were probably related to a higher abundance of Firmicutes members in addition to the hydraulic short-circuit and low abundance of Chloroflexi members. On the other hand, the fiberglass factory presented higher COD removal rates (60-83%), harboring phyla reported as the core microbiome of anaerobic digesters (Bacteroidetes, Chloroflexi, and Proteobacteria phyla). The knowledge of the structure and composition of wastewater treatment systems may provide support for the improvement of the pollutant removal in anaerobic process.

Anoxic Microbial Community Robustness Under Variation of Hydraulic Retention Time and Availability of Endogenous Electron Donors.

The ADNMED (Anaerobic Digestion, Nitrification, and Mixotrophic Endogenous Denitrification) system comprises a triple chamber configuration that was shown to provide high-quality effluent regarding carbon, nitrogen, and sulfide. Hydraulic retention time (HRT) was 7 h in the anaerobic and anoxic chambers, and 5 h in the aerobic chamber (stage A). Sewage was directly added to the anoxic chamber to provide extra organic electron donors for denitrification (stage B) to improve the nitrogen removal efficiency (stage A 47 ± 19%). The addition of sewage at a flow rate equivalent to 10% of the feed flow increased nitrogen removal efficiency to 61 ± 12%. Illumina® sequencing revealed a restructuring of the microbial community in the anoxic chamber, according to the availability of the endogenous electron donors for denitrification. At stage A, denitrification was related to the decay of biomass, while the addition of sewage during stage B stimulated the establishment of fermentative bacteria.

The use of non-adapted anaerobic consortium in batch reactors enable to couple polychlorinated biphenyl degradation and community adaptation.

The removal of polychlorinated biphenyls (PCBs) and PCB biosorption was investigated in anaerobic batch reactors with non-adapted sludge fed with 1.5 mg L-1 of six PCB congener (PCB 10, 28, 52, 153, 138 and 180), mineral medium and co-substrates. PCBs were analyzed by gas chromatography using headspace solid-phase microextraction (HS-SPME). In the methanogenic reactor the methane production, COD (Carbon Organic Demand) removal (90% of initial 2292.60 mg L-1) and consumption of volatile organic acids were verified. Nevertheless, anaerobic activity was not observed in the reactor with inactivated biomass and biosorption range of 38% to 89% was measured for distinct PCB congeners in this reactor. The PCB removal was calculated from the PCB bioavailable (not biosorbed) and reached 76% of total PCBs. The selection of some representatives of the Thermotogaceae family, Sedimentibacter and Pseudomonas at 101 days of operation in the methanogenic reactor was correlated with PCB degradation. In addition, the various removal rates for each PCB congener indicate that the removal depends on bioavailability. The selection of the former non-adapted microbiota in the methanogenic reactor combined with PCB degradation occurred at 101 days. These results allow to assert that it is possible to simultaneously couple PCB degradation and community selection, without the previous adaptation step, which is a time-consuming stage.

Evaluation of anionic surfactant removal by anaerobic degradation of commercial laundry wastewater and domestic sewage.

An expanded granular sludge bed reactor was evaluated for the anaerobic digestion of commercial laundry wastewater and domestic sewage focused on the removal of linear alkylbenzene sulfonate (LAS). The reactor was operated in three stages, all under mesophilic conditions and with a hydraulic retention time of 36 h. At stage I, the laundry wastewater was diluted with tap water (influent: 15.3 ± 4.9 mg LAS/L); at stage II, 50% of the feed volume was domestic sewage and 50% was a mixture of tap water and laundry wastewater (influent: 15.8 ± 4.9 mg LAS/L); and at stage III, only domestic sewage was used as a diluent of the laundry wastewater (influent: 24.1 ± 4.1 mg LAS/L). Due to the addition of domestic sewage the organic compounds content and LAS in the influent increased. Under such conditions, it was observed that LAS removal rate decreased from 77.2 ± 14.9% (stage I) to 55.3 ± 18.4% (stage III). Statistical tests indicated that the decrease of the LAS removal rate was significant and indicated a correlation between the removal of LAS and specific organic loading rate. The analysis of 16S rRNA gene sequencing revealed genera similar to Geobacter, Desulfovibrio, Syntrophomonas, Syntrophus, Desulfobulbus, Desulfomonile, and Desulfomicrobium, which were related to the degradation of LAS.

Proof of concept and improvement of a triple chamber biosystem coupling anaerobic digestion, nitrification and mixotrophic endogenous denitrification for organic matter, nitrogen and sulfide removal from domestic sewage.

In this study, two versions of a triple chamber biosystem, coupling anaerobic digestion, nitrification and mixotrophic endogenous denitrification (ADNMED), were evaluated and compared. They were designed to maximize the use of endogenous electron donors produced by anaerobic digestion (residual organic matter and sulfide) to abate a portion of the influent nitrogen contained in domestic sewage while removing the inconvenience of effluent sulfide. The first version was able to abate 40% of the influent nitrogen but presented operational and hydrodynamic problems, which resulted in sulfide emissions. A modified second version was proposed, improving the first approach and achieving a nitrogen abatement of more than 60% and a sulfide-free effluent, complying with local emission standards. The results demonstrated that endogenous electron donors produced by anaerobic digestion should not be neglected, and a significant cost reduction in nitrogen removal from domestic sewage could be achieved by exploiting their potential with novel reactor configurations.

Carbon-nitrogen removal in a structured-bed reactor (SBRRIA) treating sewage: Operating conditions and metabolic perspectives.

The present study evaluated the efficiency of a structured-bed reactor subjected to recirculation and intermittent aeration (SBRRIA) to promote nitrogen and carbon removal from domestic sewage. The intermittent aeration and the recycling rate of 3 keeps the desired mixing degree inside the SBRRIA. Four different operational conditions were tested by varying the hydraulic retention time (HRT) from 12 to 8 h and aerated and non-aerated periods (A/NA) from 2 h/1 h and 3 h/1 h. At the THD of 8 h and A/NA of 2 h/1 h there was a decrease in the nitrification process (77.5%) due to the increase of organic matter availability, affecting the total-N removal performance. However, by increasing the aerated period from 2 h to 3 h, the nitrification efficiency rose to 91.1%, reaching a total-N removal efficiency of 79%. The system reached a maximum total-N loading removed of 0.117 kgN.m-3.d-1 by applying an HRT of 8 h and an intermittent aeration cycle of 3 h, aerated and 1 h non-aerated. The simultaneous nitrification and denitrification (SND) process was related to a complex interplay among microorganisms affiliated mostly to Acidovorax sp., Comamonas sp., Dechloromonas sp., Hydrogenophaga sp., Mycobacterium sp., Rhodobacter sp., and Steroidobacter sp.

Anaerobic degradation of anionic surfactants by indigenous microorganisms from sediments of a tropical polluted river in Brazil.

Linear alkylbenzene sulfonate (LAS) is widely used in the formulation of domestic and industrial cleaning products, the most synthetic surfactants used worldwide. These products can reach water bodies through the discharge of untreated sewage or non-effective treatments. This study evaluates the ability of the microorganisms found in the Tiete river sediment to degrade this synthetic surfactant. The experiment was conducted in a bioreactor, operated in batch sequences under denitrifying conditions, with cycles of 24 hours and stirring at 150rpm, using 430 mL of sediments and 1 070mL of a synthetic substrate consisting of yeast extract, soluble starch, sodium bicarbonate and sucrose. LAS was added at different concentrations of l5mg/L and 30mg/L. The reactor operation was divided into the biomass adaptation to the synthetic substrate without LAS and three experimental conditions: a) addition of l5mg/L of LAS; b) 50% reduction the co-substrate concentration and 15 mg/L of LAS, and c) addition of 30mg/L of LAS and 100% co-substrate concentration. The results showed that the degradation efficiency of LAS was directly related to the addition of co-substrates and the population of denitrifying bacteria. The removal of LAS and nitrate can be achieved simultaneously in wastewater with low organic loads. The reduction in the co-substrates concentration was directly influenced by the number of denitrifying bacteria (2.2x10(13) to 1.0 x 10(8) MPN/gTVS), and consequently, LAS degradation (60.1 to 55.4%). The sediment microorganisms in the Tiete river can be used as an alternative inoculum in the treatment of wastewater with nitrate and LAS contamination.

Biodegradation of linear alkylbenzene sulfonate in commercial laundry wastewater by an anaerobic fluidized bed reactor.

The biodegradation of linear alkylbenzene sulfonate (LAS) from commercial laundry wastewater was evaluated in an anaerobic fluidized bed reactor (FBR) fed with synthetic substrate (598 mg L(-1) to 723 mg L(-1) of organic matter) supplemented with 9.5±3.1 mg L(-1) to 27.9±9.6 mg L(-1) of LAS. The average chemical oxygen demand (COD) removal efficiency was 89% and the biodegradation of LAS was 57% during the 489 days of anaerobic FBR. Higher levels of volatile fatty acids (VFA) were observed in the effluent at the stage with the best LAS removal performance. Increasing the surfactant concentration did not increase the VFA production in the effluent. The predominant VFAs after the addition of LAS were as follows: isovaleric acid and valeric acid, followed by propionic acid, caproic acid and formic acid. The similarities of 64% and 45% to Archaea and Bacteria domains were observed in the samples taken in the operating period of anaerobic FBR fed with 23.6±10 mg L(-1) and 27.9±10 mg L(-1) of LAS. During the operation stages in the reactor, Gemmatimonas, Desulfobulbus and Zoogloea were determined as the most abundant genera related to surfactant degradation using 454-Pyrosequencing.

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.

Bacterial communities in thermophilic H2-producing reactors investigated using 16S rRNA 454 pyrosequencing.

In this study, the composition and diversity of the bacterial community in thermophilic H2-producing reactors fed with glucose were investigated using pyrosequencing. The H2-producing experiments in batch were conducted using 0.5 and 2.0 g l(-1) glucose at 550 °C. Under the two conditions, the H2 production and yield were 1.3 and 1.6 mol H2 mol glucose(-1), respectively. Acetic, butyric, iso-butyric, lactic and propionic acids were detected in the two reactors. The increase in substrate concentration favored a high H2 yield. In this reactor, a predominance of acetic and iso-butyric acids, 27.7% and 40%, were measured, respectively. By means of pyrosequencing, a total of 323 and 247 operational taxonomic units were obtained, with a predominance of the phylum Firmicutes (68.73-67.61%) for reactors with 0.5 and 2.0 g l(-1) glucose, respectively. Approximately 40.55% and 62.34% of sequences were affiliated with Thermoanaerobacterium and Thermohydrogenium, microorganisms that produce H2 under thermophilic conditions.

Anaerobic degradation of anionic surfactants by indigenous microorganisms from sediments of a tropical polluted river in Brazil / Degradación anaeróbica de tensioactivos aniónicos en microorganismos autóctonos de los sedimentos de un río tropical contaminado en Brasil

Linear alkylbenzene sulfonate (LAS) is widely used in the formulation of domestic and industrial cleaning products, the most synthetic surfactants used worldwide. These products can reach water bodies through the discharge of untreated sewage or non-effective treatments. This study evaluates the ability of the microorganisms found in the Tietê river sediment to degrade this synthetic surfactant. The experiment was conducted in a bioreactor, operated in batch sequences under denitrifying conditions, with cycles of 24 hours and stirring at 150rpm, using 430mL of sediments and 1 070mL of a synthetic substrate consisting of yeast extract, soluble starch, sodium bicarbonate and sucrose. LAS was added at different concentrations of 15mg/L and 30mg/L. The reactor operation was divided into the biomass adaptation to the synthetic substrate without LAS and three experimental conditions: a) addition of 15mg/L of LAS; b) 50% reduction the co-substrate concentration and 15mg/L of LAS, and c) addition of 30mg/L of LAS and 100% co-substrate concentration. The results showed that the degradation efficiency of LAS was directly related to the addition of co-substrates and the population of denitrifying bacteria. The removal of LAS and nitrate can be achieved simultaneously in wastewater with low organic loads. The reduction in the co-substrates concentration was directly influenced by the number of denitrifying bacteria (2.2x10(13) to 1.0x10(8)MPN/gTVS), and consequently, LAS degradation (60.1 to 55.4%). The sediment microorganisms in the Tietê river can be used as an alternative inoculum in the treatment of wastewater with nitrate and LAS contamination.

El alquilbenceno sulfonato lineal (LAS) es el tensoactivo sintético más usado en todo el mundo en los produtos de limpeza domestica e industrial y puede llegar a las masas de agua a través de la descarga de aguas residuales sin tratamiento o con un tratamiento ineficaz. El objetivo del estudio consistió en evaluar la capacidad de la microbiota presente en el sedimento del río Tietê en la degradación del tensoactivo anionico - LAS. El experimento se llevó a cabo en un bioreactor de lotes secuenciales en condiciones de desnitrificación con ciclos de 24 horas, agitación de 150rpm, usando 430mL de sedimento y 1 070mL de sustrato sintético constituido por extracto de levadura, almidón soluble, bicarbonato de sodio y sacarosa. El LAS fue añadido a diferentes concentraciones de 15mg/L y 30mg/L. El funcionamiento del bioreactor se dividió en la adaptación de la biomasa con sustrato sintético sin LAS y tres condiciones experimentales: A) adición de 15mg/L de LAS; B) 15mg/L de LAS y reducción del 50% de la concentración del co-sustrato y C) 30mg/L de LAS y la concentración de 100% de co-substrato. Los resultados obtenidos muestran que la eficiencia en la degradación del LAS está directamente relacionada con la población de bacterias desnitrificadoras y que el sedimento del río Tietê se puede utilizar como inóculo en el tratamiento de LAS en condiciones desnitrificadoras. La población de bacterias fue capaz de degradar el LAS independiente de la fuente de carbón adicionada. La remoción de LAS y de nitrato se puede lograr simultáneamente en aguas residuales con una baja carga orgánica. La reducción de la concentración del co-sustrato fue influenciado directamente por la población de bacterias desnitrificantes (2.2x10(13) a 1.0x10(8)MNP/gTVS) y por lo tanto la degradación de LAS (60.1-55.4%). Los microorganismos en el sedimento del río Tietê se pueden usar como inóculo alternativo para el tratamiento de efluentes contaminados con nitrato y LAS.

Microbial characterization and degradation of linear alkylbenzene sulfonate in an anaerobic reactor treating wastewater containing soap powder.

The aim of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) in an anaerobic fluidized bed reactor (AFBR) treating wastewater containing soap powder as LAS source. At Stage I, the AFBR was fed with a synthetic substrate containing yeast extract and ethanol as carbon sources, and without LAS; at Stage II, soap powder was added to this synthetic substrate obtaining an LAS concentration of 14 ± 3 mg L(-1). The compounds of soap powder probably inhibited some groups of microorganisms, increasing the concentration of volatile fatty acids (VFA) from 91 to 143 mg HAc L(-1). Consequently, the LAS removal rate was 48 ± 10% after the 156 days of operation. By sequencing, 16S rRNA clones belonging to the phyla Proteobacteria and Synergistetes were identified in the samples taken at the end of the experiment, with a remarkable presence of Dechloromonas sp. and Geobacter sp.

Impact of inocula and operating conditions on the microbial community structure of two anammox reactors.

The microbial community structure of the biomass selected in two distinctly inoculated anaerobic oxidation of ammonium (anammox) reactors was investigated and compared with the help of data obtained from 454-pyrosequencing analyses. The anammox reactors were operated for 550 days and seeded with different sludges: sediment from a constructed wetland (reactor I) and biomass from an aerated lagoon part of the oil-refinery wastewater treatment plant (reactor II). The anammox diversity in the inocula was evaluated by 16S rRNA gene-cloning analysis. The diversity of anammox bacteria was greater in the sludge from the oil-refinery (three of the five known genera of anammox were detected) than in the wetland sludge, in which only Candidatus Brocadia was observed. Pyrosequencing analysis demonstrated that the community enriched in both reactors had differing compositions despite the nearly similar operational conditions applied. The dominant phyla detected in both reactors were Proteobacteria, Chloroflexi, Planctomycetes, and Acidobacteria. The phylum Bacteroidetes, which is frequently observed in anammox reactors, was not detected. However, Acidobacteria and GN04 phyla were observed for the first time, suggesting their importance for this process. Our results suggest that, under similar operational conditions, anammox populations (Ca. Brocadia sinica and Ca. Brocadia sp. 40) were selected in both reactors despite the differences between the two initial inocula. Taken together, these results indicated that the type of inoculum and the culture conditions are key determinants of the general microbial composition of the biomass produced in the reactors. Operational conditions alone might play an important role in anammox selection.

Influence of volatile fatty acid concentration stability on anaerobic degradation of linear alkylbenzene sulfonate.

Linear alkylbenzene sulfonate (LAS) is an anionic surfactant used in cleaning products, which is usually found in wastewaters. Despite the greater LAS removal rate related to a lower concentrations of volatile fatty acids (VFA), the influence of different ranges of VFA on LAS degradation is not known. LAS degradation was evaluated in upflow anaerobic sludge blanket (UASB) and expanded granular sludge bed (EGSB) reactors at different ranges of VFA concentrations. The reactors were fed with a synthetic wastewater containing LAS (14 mg/L). A greater LAS removal rate (40-80%) was related to the lower and narrower range of acetic acid concentration (1-22 mg/L) in the EGSB reactor. In the UASB reactor, the acetic acid concentrations presented a wider range (2-45 mg/L), and some low LAS removal rates (around 20-25%) were observed even at low acetic acid concentrations (<10 mg/L). The high recirculation rate in the EGSB reactor improved substrate-biomass contact, which resulted in a narrower range of VFA and greater LAS removal rate.

Optimization of linear alkylbenzene sulfonate (LAS) degradation in UASB reactors by varying bioavailability of LAS, hydraulic retention time and specific organic load rate.

Degradation of linear alkylbenzene sulfonate (LAS) in UASB reactors was optimized by varying the bioavailability of LAS based on the concentration of biomass in the system (1.3-16 g TS/L), the hydraulic retention time (HRT), which was operated at 6, 35 or 80 h, and the concentration of co-substrates as specific organic loading rates (SOLR) ranging from 0.03-0.18 g COD/g TVS.d. The highest degradation rate of LAS (76%) was related to the lowest SOLR (0.03 g COD/g TVS.d). Variation of the HRT between 6 and 80 h resulted in degradation rates of LAS ranging from 18% to 55%. Variation in the bioavailability of LAS resulted in discrete changes in the degradation rates (ranging from 37-53%). According to the DGGE profiles, the archaeal communities exhibited greater changes than the bacterial communities, especially in biomass samples that were obtained from the phase separator. The parameters that exhibited more influence on LAS degradation were the SOLR followed by the HRT.

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.