Biomethane recovery through co-digestion of cheese whey and glycerol in a two-stage anaerobic fluidized bed reactor: Effect of temperature and organic loading rate on methanogenesis.

Anaerobic digestion for CH4 recovery in wastewater treatment has been carried out with different strategies to increase process efficiency, among which co-digestion and the two-stage process can be highlighted. In this context, this study aimed at evaluating the co-digestion of cheese whey and glycerol in a two-stage process using fluidized bed reactors, verifying the effect of increasing the organic loading rate (OLR) (2-20 g-COD.L-1.d-1) and temperature (thermophilic and mesophilic) in the second stage methanogenic reactor. The mesophilic methanogenic reactor (R-Meso) (mean temperature of 22 °C) was more tolerant to high OLR and its best performance was at 20 g-COD.L-1.d-1, resulting in methane yield (MY) and methane production (MPR) of 273 mL-CH4.g-COD-1 and 5.8 L-CH4.L-1.d-1 (with 67% of CH4), respectively. Through 16S rRNA gene massive sequencing analysis, a greater diversity of microorganisms was identified in R-Meso than in R-Thermo (second stage methanogenic reactor, 55 °C). Firmicutes was the phyla with higher relative abundance in R-Thermo, while in R-Meso the most abundant ones were Proteobacteria and Bacteroidetes. Regarding the Archaea domain, a predominance of hydrogenotrophic microorganisms could be observed, being the genera Methanothermobacter and Methanobacterium the most abundant in R-Thermo and R-Meso, respectively. The two-stage system composed with a thermophilic acidogenic reactor + R-Meso was more adequate for the co-digestion of cheese whey and glycerol than the single-stage process, promoting increases of up to 47% in the energetic yield (10.3 kJ.kg-COD-1) and 14% in organic matter removal (90.5%).

Statistical optimization of methane production from brewery spent grain: Interaction effects of temperature and substrate concentration.

This study evaluated the effects of thermal pretreatment of brewery spent grain (BSG) (by autoclave 121 °C, 1.45 atm for 30 min) on methane production (CH4). Operation temperature (31-59 °C) and substrate concentration (8.3-19.7 g BSG.L-1) factors were investigated by Response Surface Methodology (RSM) and Central Composite Design (CCD). Values ranging from 81.1 ± 2.0 to 290.1 ± 3.5 mL CH4.g-1 TVS were obtained according to operation temperature and substrate concentration variation. The most adverse condition for methanogenesis (81.1 ± 2.0 mL CH4.g-1 TVS) was at 59 °C and 14 g BSG.L-1, in which there was increase in the organic matter concentration from 173.6 ± 4.94 to 3036 ± 7.78 mg.L-1) result of a higher final concentration of volatile fatty acids (VFA, 2662.7 mg.L-1). On the other hand, the optimum condition predicted by the statistical model was at 35 °C and 18 g BSG.L-1 (289.1 mL CH4.g-1 TVS), which showed decrease in the organic matter concentration of 78.6% and a lower final concentration of VFA (533.2 mg.L-1). Hydrogenospora and Methanosaeta were identified in this optimum CH4 production condition, where acetoclastic methanogenic pathway prevailed. The CH4 production enhancement was concomitant to acetic acid concentration decrease (from 578.9 to 135.7 mg.L-1).

Microbial and functional characterization of an allochthonous consortium applied to hydrogen production from Citrus Peel Waste in batch reactor in optimized conditions.

Energy recovery from lignocellulosic waste has been studied as an alternative to the problem of inappropriate waste disposal. The present study aimed at characterizing the microbial community and the functional activity of reactors applied to H2 production through lignocellulosic waste fermentation in optimized conditions. The latter were identified by means of Rotational Central Composite Design (RCCD), applied to optimize allochthonous inoculum concentration (2.32-5.68 gTVS/L of granular anaerobic sludge), pH (4.32-7.68) and Citrus Peel Waste (CPW) concentration (1.55-28.45 g/L). After validation, the conditions identified for optimal H2 production were 4 gSTV/L of allochthonous inoculum, 29.8 g/L of CPW (substrate) and initial pH of 8.98. In these conditions, 48.47 mmol/L of H2 was obtained, which is 3.64 times higher than the concentration in unoptimized conditions (13.31 mmol H2/L using 15 g/L of CPW, 2 gTVS/L of allochthonous inoculum, pH 7.0). Acetogenesis was the predominant pathway, and maximal concentrations of 3,731 mg/L of butyric acid and 3,516 mg/L of acetic acid were observed. Regarding the metataxonomic profile, Clostridium genus was dramatically favored in the optimized condition (79.78%) when compared to the allochthonous inoculum (0.43%). It was possible to identify several genes related to H2 (i.e dehydrogenases) and volatile fatty acids (VFA) production and with cellulose degradation, especially some CAZymes from the classes Auxiliary Activities, Glycoside Hydrolases and Glycosyl Transferase. By means of differential gene expression it was observed that cellulose degradation and acetic acid production pathways were overabundant in samples from the optimized reactors, highlighting endo-ß-1,4-glucanase/cellulose, endo-ß-1,4-xylanase, ß-glucosidase, ß-mannosidase, cellulose ß-1,4-cellobiosidase, cellobiohydrolase, and others, as main the functions.

Orange Bagasse Pellets as a Carbon Source for Biobutanol Production.

Due to the environmental concerns, the conversion of lignocellulosic waste can be the key to produce bioproducts and biofuels such as butanol. This study aimed to present and evaluate orange bagasse pellets (OBP) as a carbon source to produce butan-1-ol production via ABE fermentation using Clostridium beijerinckii. These bagasse pellets were characterized, holocellulose (18.99%), alfacellulose (5.37%), hemicellulose (13.62%), lignin (6.16%), pectin (7.21%), protein (3.14%), and was tested under three different pretreatments, which were the following: (a) ultrasound, (b) autohydrolysis, and (c) acid-diluted hydrolysis followed by enzymatic hydrolysis to verify an amount of fermentable total reducing sugars. ANOVA was used and pretreatments followed by enzymatic hydrolysis do not enhance a significant amount of available sugars compared to raw bagasse. The ABE fermentation was carried out in batch reactors at 37 °C under agitation of 160 rpm and anaerobic conditions, using OBP without treatment followed by enzymatic hydrolysis. Using a non-mutant microorganism, the fermentation achieved butyric acid yields of 3762.68 mg L-1 for control and 2488.82 mg L-1 for OBP and the butanol production was 63.86 mg L-1 and 196.80 mg L-1 for OBP and the control (glucose) assay, respectively. The results of this solvent's production have shown that OBP has the potential for ABE fermentation and a promising feedstock.

The influence of upflow velocity and hydraulic retention time changes on taxonomic and functional characterization in Fluidized Bed Reactor treating commercial laundry wastewater in co-digestion with domestic sewage.

A large-scale (19.8L) Fluidized Bed Reactor (FBR) operated for 592 days was used to assess the removal performance of linear alkylbenzene sulfonate (LAS). Adjustments in hydraulic retention time (HRT) (18 and 30 h), ethanol (50, 100, 200 mg L-1) and linear alkylbenzene sulfonate (LAS) concentration (6.3-24.7 mg L-1) with taxonomic and functional characterization of biomass using Whole Genome Shotgun Metagenomic (WGSM) represented a major step forward for optimizing biological treatments of LAS. In addition, the variation of the upflow velocity (0.5, 0.7 and 0.9 cm s-1) was investigated, which is a parameter that had not yet been correlated with the possibilities of LAS removal in FBR. Lower Vup (0.5 cm s-1) allied to higher ethanol concentration (200 mg L-1) resulted in lower LAS removal (29%) with predominance of methanogenic archaea and genes related to methanogenesis, while higher Vup (0.9 cm s-1) led to aerobic organisms and oxidative phosphorylation genes. An intermediate Vup (0.7 cm s-1) and higher HRT (30 h) favored sulfate reducing bacteria and genes related to sulfur metabolism, which resulted in the highest LAS (83%) and COD (77%) removal efficiency.

Controlling methane and hydrogen production from cheese whey in an EGSB reactor by changing the HRT.

This study assessed the effects of hydraulic retention time (HRT; 8 h-0.25 h) on simultaneous hydrogen and methane production from cheese whey (5000 mg carbohydrates/L) in a mesophilic (30 °C) expanded granular sludge bed (EGSB) reactor. Methane production was observed at HRTs from 4 to 0.25 h. The maximum methane yield (9.8 ± 1.9 mL CH4/g CODap, reported as milliliter CH4 per gram of COD applied) and methane production rate (461 ± 75 mL CH4/day Lreactor) occurred at HRTs of 4 h and 2 h, respectively. Hydrogen production increased as methane production decreased with decreasing HRT from 8 to 0.25 h. The maximum hydrogen yield of 3.2 ± 0.3 mL H2/g CODap (reported as mL H2 per gram of COD applied) and hydrogen production rate of 1951 ± 171 mL H2/day Lreactor were observed at the HRT of 0.25 h. The decrease in HRT from 8 to 0.25 h caused larger changes in the bacterial populations than the archaea populations. With the decrease in HRT (6 h-0.25 h), the Shannon diversity index decreased (3.02-2.87) for bacteria and increased (1.49-1.83) for archaea. The bacterial dominance increased (0.059-0.066) as the archaea dominance decreased (0.292-0.201) with the HRT decrease from 6 to 0.25 h.

4-Nonylphenol degradation changes microbial community of scale-up Anaerobic Fluidized Bed Reactor.

Nonylphenol Ethoxylate (NPe) is a nonionic surfactant widely applied in domestic and industrial uses and its degradation generates the endocrine disruptor 4-Nonylphenol (4-NP). The effects of this compound in biological sewage treatment are uncertain, especially in anaerobic systems. The aim of this study was to assess the 4-NP removal and degradation in scale-up (20 L) Anaerobic Fluidized Bed Reactor (AFBR) filled with sand as support material, operated with Hydraulic Retention Time (HRT) of 18 h, fed with synthetic sewage plus 4-NP, performed in four phases named Phase I (894 mg COD L-1), Phase II (878 mg COD L-1, 127 µg 4-NP L-1), Phase III (940 mg COD L-1, 270 µg 4-NP L-1) and Phase IV (568 mg COD L-1, 376 µg 4-NP L-1). 4-NP did not affect reactor stability and organic matter removal remained stable at 94%. Highest 4-NP removal (78%) occurred for highest 4-NP influent (Phase IV), which resulted from biomass adaptation in the presence of ethanol. Through the 4-NP total mass balance, about 70% was biodegraded and 1% adsorbed on the sand bed. 4-NP addition promoted selection of microbial consortium strongly linked to aromatic compounds and surfactants degradation such as Geothrix, Holophaga, Aeromonas, Pelobacter, Pseudomonas, Delftia.

Dynamics and response of microbial diversity to nutritional conditions in denitrifying bioreactor for linear alkylbenzene sulfonate removal.

The aim of this study was to evaluate the microbial structure and phylogenetic diversity under the influence of nutritional conditions and hydraulic retention time (HRT) in fluidized bed reactors (FBR), operated in short HRT (8 h - FBR8; 12 h - FBR12) for linear alkylbenzene sulfonate (LAS) removal from laundry wastewater. After each phase, biofilm samples from FBR8 and FBR12 were submitted to microbial sequencing by Mi-Seq Illumina®. Higher LAS removal rates were observed after 313 days, achieving 99 ± 3% in FBR12 (22.5 ± 5.9 mg LAS/L affluent) and 93 ± 12% in FBR8 (20.6 ± 4.4 mg LAS/L affluent). Different modifications involving genera of bacteria were observed throughout the reactors operation. The identified microorganisms were, mostly, related to LAS degradation and nitrogen conversion such as Dechloromonas, Flavobacterium, Pseudomonas, and Zoogloea.

Bacillus sp. isolated from banana waste and analysis of metabolic pathways in acidogenic systems in hydrogen production.

The autochthonous bacterial strain was isolated from banana waste (BW) for hydrogen production and organic acids from different pure substrates and BW. The potential production of H2 and metabolic pathways were analyzed at pH 7.0 at 37 °C. Facultative anaerobic bacterium similar to Bacillus sp. was identified, with generation time (Tg) and growth rate (µ) of 0.43 h and 1.60 h-1, respectively, from glucose. The hydrogen production (P) using pure substrates was observed between 10.81 mmol.L-1 and 17.75 mmol.L-1 from xylose and maltose, respectively. The biggest and smallest hydrogen P of 18.77 mmol.L-1 and 1.72 mmol.L-1, were obtained with 3.5 and 0.5 g.L-1 of cellobiose, respectively. The highest hydrogen yield (126.93 mL.g-1carbohydrates added) was obtained with 2 g.L-1 cellobiose. In the assay using banana waste (5 g.L-1) the maximum P and yield (YH2) of 31.7 mmol.L-1 and 94.66 mL H2 g-1carbohydrates added, was obtained respectively. The main metabolic pathway of hydrogen production by Bacillus sp. RM1 from banana waste was acetic-butyric acid of 487.69 g.L-1 and 535.88 g.L-1, respectively. The accessibility of various carbon sources by Bacillus sp. RM1 in fermentation can benefit the hydrogen production from complex organic substrates in the bioaugmentation process.

Optimization of hydrogen and organic acids productions with autochthonous and allochthonous bacteria from sugarcane bagasse in batch reactors.

The individual and mutual effects of substrate concentration (from 0.8 to 9.2 g/L) and pH (from 4.6 to 7.4) on hydrogen and volatile fatty acids production from sugarcane bagasse (SCB) were investigated in batch reactors, using a response surface methodology (RSM) and central composite design (CCD). The maximum of 23.10 mmoL H2/L was obtained under optimized conditions of 7.0 g SCB/L and pH 7.2, at 37 °C through the acetic acid pathway (1.57 g/L). Butyric and succinic acids were the major volatile fatty acids (VFA) produced in the fermentation process (from 0.66 to 1.88 g/L and from 1.06 to 1.65 g/L, respectively). According to the results, the RSM and CCD were useful tools to achieve high hydrogen production rates using Clostridium, Bacillus and Enterobacter, identified by Illumina sequencing (16S RNAr) in the fermentative consortium, and Clostridium and Paenibacillus, autochthonous bacteria from SCB. Significant changes were observed in the microbial community according to the changes in the independent variables, since the genera in the central point condition (5.0 g SCB/L and pH 6.0) were Lactobacillus, Escherichia and Clostridium, and Bacteroides and Enterobacter, which were identified in the optimized condition (7.0 g SCB/L and pH 7.2).

Design and optimization of hydrogen production from hydrothermally pretreated sugarcane bagasse using response surface methodology.

Hydrogen production from hydrothermally pretreated (200 °C for 10 min at 16 bar) sugarcane bagasse was analyzed using response surface methodology. The yeast extract concentration and the temperature had a significant influence for hydrogen production (p-value 0.027 and 0.009, respectively). Maximum hydrogen production (17.7 mmol/L) was observed with 3 g/L yeast extract at 60 °C (C10). In this conditions were produced acetic acid (50.44 mg/L), butyric acid (209.71 mg/L), ethanol (38.4 mg/L), and methane (6.27 mmol/L). Lower hydrogen productions (3.5 mmol/L and 3.9 mmol/L) were observed under the conditions C7 (2 g/L of yeast extract, 35.8 °C) and C9 (1 g/L of yeast extract, 40 °C), respectively. The low yeast extract concentration and low temperature caused a negative effect on the hydrogen production. By means of denaturing gradient gel electrophoresis 20% of similarity was observed between the archaeal population of mesophilic (35 and 40 °C) and thermophilic (50, 60 and 64 °C) reactors. Likewise, similarity of 22% was noted between the bacterial population for the reactors with the lowest hydrogen production (3.5 mmol/L), at 35.8 °C and with the highest hydrogen production (17.7 mmol/L) at 60 °C demonstrating that microbial population modification was a function of incubation temperature variation.

Bioconversion of waste office paper to hydrogen using pretreated rumen fluid inoculum.

In this study, a microbial consortium from an acid-treated rumen fluid was used to improve the yields of H2 production from paper residues in batch reactors. The anaerobic batch reactors, which contained paper and cellulose, were operated under three conditions: (1) 0.5 g paper/L, (2) 2 g paper/L, and (3) 4 g paper/L. Cellulase was added to promote the hydrolysis of paper to soluble sugars. The H2 yields were 5.51, 4.65, and 3.96 mmol H2/g COD, respectively, with substrate degradation ranging from 56 to 65.4 %. Butyric acid was the primary soluble metabolite in the three reactors, but pronounced solventogenesis was detected in the reactors incubated with increased paper concentrations (2.0 and 4.0 g/L). A substantial prevalence of Clostridium acetobutylicum (99 % similarity) was observed in the acid-treated rumen fluid, which has been recognized as an efficient H2-producing strain in addition to ethanol and n-butanol which were also detected in the reactors.

Soil contamination assessment for Pb, Zn and Cd in a slag disposal area using the integration of geochemical and microbiological data.

Improper disposal of mining waste is still considered a global problem, and further details on the contamination by potentially toxic metals are required for a proper assessment. In this context, it is important to have a combined view of the chemical and biological changes in the mining dump area. Thus, the objective of this study was to evaluate the Pb, Zn and Cd contamination in a slag disposal area using the integration of geochemical and microbiological data. Analyses of soil organic matter (SOM), pH, Eh, pseudo-total concentration of metals, sequential extraction and microbial community by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) were conducted. Metal availability was evaluated based on the geoaccumulation index (I geo), ecological risk ([Formula: see text]), Risk Assessment Code (RAC) and experimental data, and different reference values were tested to assist in the interpretation of the indices. The soil pH was slightly acidic to neutral, the Eh values indicated oxidized conditions and the average SOM content varied from 12.10 to 53.60 g kg-1. The average pseudo-total concentrations of metals were in the order of Zn > Pb > Cd. Pb and Zn were mainly bound to the residual fraction and Fe-Mn oxides, and a significant proportion of Cd was bound to the exchangeable and carbonate fractions. The topsoil (0-20 cm) is highly contaminated (I geo) with Cd and has a very high potential ecological risk ([Formula: see text]). Higher bacterial diversity was mainly associated with higher metal concentrations. It is concluded that the integration of geochemical and microbiological data can provide an appropriate evaluation of mining waste-contaminated areas.

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.

Production of H(2) from cellulose by rumen microorganisms: effects of inocula pre-treatment and enzymatic hydrolysis.

H2 production from cellulose, using rumen fluid as the inoculum, has been investigated in batch experiments. Methanogenic archaea were inhibited by acid pre-treatment, which also inhibited cellulolytic microorganisms, and in consequence, the conversion of cellulose to H2. Positive results were observed only with the addition of cellulase. H2 yields were 18.5 and 9.6 mmol H2 g cellulose(-1) for reactors with 2 and 4 g cellulose l(-1) and cellulase, respectively. H2 was primarily generated by the butyric acid pathway and this was followed by formation of acetic acid, ethanol and n-butanol. In reactors using 4 g cellulose l(-1) and cellulase, the accumulation of alcohols negatively affected the H2 yield, which changed the fermentation pathways to solventogenesis. PCR-DGGE analysis showed changes in the microbial communities. The phylogenetic affiliations of the bands of DGGE were 99 % similar to Clostridium sp.

Potential methanogenic and degradation of nonylphenol ethoxylate from domestic sewage: unravelling the essential roles of nutritional conditions and microbial community.

Nonylphenol ethoxylathe (NPEO) is a non-ionic surfactant of increasing concern, used in the formulation of laundry detergents and is frequently found in aquatic environments. The purpose of this study was to evaluate the effects of yeast extract (YE) and sodium fumarate (SF) in NPEO removal from domestic sewage under anaerobic conditions via central composite rotatable design (CCRD) and response surface methodology (RSM). Experiments were designed by varying concentrations of NPEO (1.6-5.8 mg L-1), YE (131.8-468.2 mg L-1) and SF (97.7-602.3 mg L-1) in batch reactors. SF and YE addition significantly influenced NPEO removal and CH4 production. Optimal values of YE (400 mg L-1) and SF (200 mg L-1) result in removal efficiency of 97% for 5 mg L-1 of NPEO, being mostly removed by biodegradation (86%). Meanwhile COD removal was 95% and methane yield was 134 ± 4 NmLCH4 g-¹CODremoved. The most abundant Bacteria genus identified were Macellibacteroides, Longilinea, Petrimonas and Proteiniphilum, while for Archaea, Methanosaeta and Methanoregula were the genera identified in higher relative abundances in optimized conditions.

Effects of capping on microbial populations and contaminant immobilization in an old unlined landfill.

This research aimed at evaluating the effects of capping on the mitigation of impacts generated by a closed unlined landfill in São Carlos, SP, Brazil. Physicochemical and microbiological analyses (16S rRNA sequencing) of buried solid waste samples were performed, in capped and uncapped areas. Even though leachate pockets could still be encountered in capped areas, the capping construction reduced oxygen availability and created more reducing conditions, propitiating the development of sulfate-reducing bacteria and possibly contributing to the precipitation of the metals Pb, Cd, Ni, Co, As, and Zn as metal sulfides, causing their immobilization. The microbial populations adapted to the anaerobic conditions created under capped zones belonged to the phyla Firmicutes, Chloroflexi, and Euryarchaeota and the genera Methanosaeta, Hydrogenispora, Smithella, and Gelria. Differently, the phyla Acidobacteria, Proteobacteria, Bacteroidetes, and Actinobacteria were more abundant in samples from the uncapped zones, in which the abundance of different genera varied homogeneously. Methanogenic activity was not impaired by the intervention measure, as assessed by the specific methanogenic activity (SMA). Capping of old unlined landfills brings benefits to the immobilization of metals and does not impair microbial degradation, being effective for the mitigation of impacts on soils and water resources.

Microbial and functional characterization of granulated sludge from full-scale UASB thermophilic reactor applied to sugarcane vinasse treatment.

Considering the scarcity of data in the literature regarding phylogenetic and metabolic composition of different inocula, especially those from thermophilic conditions, this research aimed at characterizing the microbial community and preferable metabolic pathways of an UASB reactor sludge applied to the thermophilic treatment (55°C) of sugarcane vinasse, by means of shotgun metagenomics. After its metabolic potential was depicted, it was possible to observe several genes encoding enzymes that are of great importance to anaerobic digestion processes with different wastes as substrate, especially regarding the biodegradation of carbohydrates and ligninolytic compounds, glycerolypids, volatile fatty acids and alcohols metabolism and biogas (H2 and CH4) production. The genera identified in higher relative abundances for Bacteria domain were Sulfirimonas (37.52 ± 1.8%), possibly related to the sludge endogenic activity due to its strong relation with a peptidoglycan lyase enzymes family, followed by Fluviicola (5.01 ± 1.0%), Defluviitoga (4.36 ± 0.2%), Coprothermobacter (4.32 ± 0.5%), Fervidobacterium (2.93 ± 0.3%), Marinospirillum (2.75 ± 0.2%), Pseudomonas (2.14 ± 0.2%) and Flavobacterium (1.78 ± 0.1%), mostly related with carbohydrates fermentations and/or H2 production. For Archaea domain, Methanosarcina (0.61 ± 0.1%), Methanothermobacter (0.38 ± 0.0%), Methanoculleus (0.30 ± 0.1%), Thermococcus (0.03 ± 0.0%), Methanolobus (0.02 ± 1.8%), Methanobacterium (0.013 ± 0.0%), Aciduliprofundum and Pyrococcus (0.01 ± 0.0%) were the most dominant ones, being Methanosarcina the most related with methanogenesis. It was concluded that the robust inoculum description performed in this study may subside future biotechnological researches by using similar inocula (UASB sludges), focusing on the obtainment of value-added by-products by means of anaerobic digestion, such as volatile fatty acids, alcohols and biogas (H2 and CH4), by using several types of waste as substrate.

Microbial diversity and metabolic inference of diclofenac removal in optimised batch heterotrophic-denitrifying conditions by means of factorial design.

Using the Response Surface Methodology (RSM) and Rotational Central Composite Design (RCCD), this study evaluated the removal of DCF under denitrifying conditions, with ethanol as cosubstrate, in batch reactors, being 1 L Erlenmeyer flasks (330 mL of reactional volume) containing Dofing medium and kept under agitation at 130 rpm and incubated at mesophilic temperature (30 °C). It considered the individual and multiple effects of the variables: nitrate (130 - 230 mg NO3- L-1), DCF (60-100 µg DCF L-1) and ethanol (130 - 230 mg EtOH L-1). The highest drug removal efficiency (17.5%) and total nitrate removal were obtained at 176.6 ± 4.3 mg NO3 -L-1, 76.8 ± 3.7 µg DCF L-1, and 180.0 ± 2.5 mg EtOH L-1. Under such conditions, the addition of ethanol and nitrate was significant for the additional removal of diclofenac (p > 0.05). The prevalence of Rhodanobacter, Haliangium and Terrimonas in the inoculum biomass (activated sludge systems) was identified through the 16S rRNA gene sequencing. The potential of these genera to remove nitrate and degrade diclofenac was inferred, and the main enzymes potentially involved in this process were α-methylacyl-CoA racemase, long-chain fatty acid-CoA ligase, catalases and pseudoperoxidases.

Two problems in one shot: Vinasse and glycerol co-digestion in a thermophilic high-rate reactor to improve process stability even at high sulfate concentrations.

Anaerobic co-digestion (AcoD) of sugarcane vinasse and glycerol can be profitable because of the destination of two biofuel wastes produced in large quantities in Brazil (ethanol and biodiesel, respectively) and the complementary properties of these substrates. Thus, the objective of this study was to assess the effect of increasing the organic loading rate (OLR) from 2 to 20 kg COD m-3 d-1 on the AcoD of vinasse and glycerol (50 %:50 % on a COD basis) in a thermophilic (55 °C) anaerobic fluidized bed reactor (AFBR). The highest methane production rate was observed at 20 kg COD m-3 d-1 (8.83 L CH4 d-1 L-1), while the methane yield remained stable at around 265 NmL CH4 g-1 CODrem in all conditions, even when influent vinasse reached 1811 mg SO42- L-1 (10 kg COD m-3 d-1). Sulfate was not detected in the effluent. Bacterial genera related to sulfate removal, such as Desulfovibrio and Desulfomicrobium, were observed by means of shotgun metagenomic sequencing at 10 kg COD m-3 d-1, as well as the acetoclastic archaea Methanosaeta and prevalence of genes encoding enzymes related to acetoclastic methanogenesis. It was concluded that process efficiency and methane production occurred even in higher sulfate concentrations due to glycerol addition.