Effect of loading rate and pH on glycerol fermentation and microbial population in an upflow anaerobic filter reactor.

A reactor with silicone tubes as support medium was used for glycerol fermentation. The experimental set-up consisted of three phases. In P1, the applied glycerol loading rate (gly-LR) was in the range of 6-10 g.L-1.d-1 at an influent pH of 7.9 ± 0.4. In P2, gly-LR was kept constant (18.0 ± 1.8 g.L-1.d-1) with different doses of NaHCO3. Finally in P3, two different gly-LR (9 and 18 g.L-1.d-1) were evaluated, dosing 1 g-NaHCO3 per g-COD of glycerol. Glycerol consumption was close 90%. The main end-product was 1,3-propanediol (1,3-PDO) (0.40 mol.mol-gly-1), but ethanol was also generated, particularly at pH above 8 and low gly-LR (0.20 mol.mol-gly-1). After 1-year operation with glycerol as the only carbon source, a drastic shift in the bacterial community was observed. The 1,3-PDO producers Lacrimispora and Clostridium became dominant, although non-glycerol-degrading fermentative genera, e.g., Actinomyces and Eubacterium, thrived at the expense of cellular breakdown products.

Effect of individual or combined physical and chemical factors on the anaerobic biodegradation of linear alkylbenzene sulphonate.

The effect of six important factors on the anaerobic biodegradation of linear alkylbenzene sulphonate (LAS) was evaluated using a response surface methodology. The factors were: (i) co-substrate concentration (CC), (ii) contact time between LAS and microorganisms, (iii) temperature, (iv) hardness, (v) pH, and (vi) LAS source. The results showed that individually or combined, CC with chemical oxygen demand (COD) ≤50 mg L-1 was the factor that mostly favoured LAS biodegradation; whereas at COD >50 mg L-1, adsorption to sludge and solubilisation in the aqueous medium were favoured. Two-factor interactions promoted the highest percentages of biodegradation (45-52%), adsorption (43-45%), and solubilisation (18-25%). The three-factor interactions resulted in small percentage increases of up to 11%, 5%, and 13% for biodegradation, adsorption, and solubilisation, respectively, compared to those of two-factor interactions. The interactions of four, five, and six factors resulted in a non-significant effect on LAS biodegradation, adsorption, and solubilisation, with percentages close to those quantified for the two- and three-factor interactions. Concentrations of up to 30 mg LAS L-1 did not significantly affect the COD removal efficiency (74-88%) from the medium. These values are commonly obtained in full-scale anaerobic systems used to treat domestic sewage.

Selecting the best electron donor and operational temperature for the rapid biotransformation of the insensitive munitions compound 2,4-dinitroanisole (DNAN) by anaerobic sludge.

2,4-Dinitroanisole (DNAN) is a toxic compound increasingly used by the military that can be released into the environment on the soil of training fields and in the wastewater of manufacturing plants. DNAN's nitro groups are anaerobically reduced to amino groups by microorganisms when electron donors are available. Using anaerobic sludge as the inoculum, we tested different electron donors for DNAN bioreduction at 20 and 30 °C: acetate, ethanol, pyruvate, hydrogen, and hydrogen + pyruvate. Biotic controls without external electron donors and abiotic controls with heat-killed sludge were also assayed. No DNAN conversion was observed in the abiotic controls. In all biotic treatments, DNAN was reduced to 2-methoxy-5-nitroaniline (MENA), which was further reduced to 2,4-diaminoanisole (DAAN). Ethanol or acetate did not increase DNAN reduction rate compared to the endogenous control. The electron donors that caused the fastest DNAN reductions were (rates at 30 °C): H2 and pyruvate combined (311.28 ± 10.02 µM·d-1·gSSV-1), followed by H2 only (207.19 ± 5.95 µM·d-1·gSSV-1), and pyruvate only (36.35 ± 2.95 µM·d-1·gSSV-1). Raising the temperature to 30 °C improved DNAN reduction rates when pyruvate, H2, or H2 + pyruvate were used as electrons donors. Our results can be applied to optimize the anaerobic treatment of DNAN-containing wastewater.

The effect of methanogenesis inhibition, inoculum and substrate concentration on hydrogen and carboxylic acids production from cassava wastewater.

Manipueira is a carbohydrate-rich agro-industrial waste from cassava processing. It is considered well suitable for biotechnological processes, such as hydrogen and carboxylic acids production, due to the high content of easily degradable organic matter. However, the proper methanogenesis inhibition method, inoculum type, and organic loads are factors still limiting the processes. The objective in this work was to evaluate the effects of such factors on byproducts production in anaerobic reactors. Batch experiments were conducted with 2.3-L flasks during two operational phases. In the first phase (P1), inhibition of methanogens in the sludge was evaluated using acetylene (1% v/v of headspace) and heat treatment (120 °C, 1 atm for 30 min). In the second phase (P2), three inoculum types obtained from common anaerobic sludges (bovine rumen and sludges from municipal and textile industrial wastewater treatment plants) were individually assayed. P2 aimed to identify the best inoculum, based on hydrogen production ability, which was tested for three initial concentrations of manipueira in terms of chemical oxygen demand (COD) (10, 20 and 40 g O2/L). Results of P1 indicated that either acetylene or heat treatment efficiently inhibited methanogenesis, with no methane production. However, the maximum H2 production potential by applying heat treatment (~ 563 mL) was more than twice compared with that by acetylene treatment (~ 257 mL); and butyrate was the main carboxylic acid by-product (~ 3 g/L). In P2 experiments after sludge heat treatment, the highest hydrogen yield (1.66 ± 0.07 mol H2/mol glucose) and caproic acid production (~ 2 g/L) were observed at 20 g O2/L of manipueira COD, when bovine rumen was the inoculum. The primary metabolic degradation products in all P2 experiments were ethanol, acetic, butyric, propionic and caproic acids. The finding of caproic acid detection indicated that the applied conditions in manipueira anaerobic degradation favored carbon chain elongation over methanogenesis.

Effects of the operational conditions on the production of 1,3-propanediol derived from glycerol in anaerobic granular sludge reactors.

The aim of this study has been to produce 1,3-propanediol (1,3-PDO) from glycerol (gly) fermentation by means of a microbial mixed culture (granular sludge), as well as to establish the operational conditions of two up-flow anaerobic sludge blanket (UASB) reactors in order to achieve a maximum 1,3-PDO yield. The UASB reactors with initial pH values set at 6.8 and 5.5 were operated at 30 °C during 165 days. Thirteen variables were previously screened by a Plackett-Burman (PB) design; results showed that yeast extract, MgSO4 and methanogenesis inhibition (by heat shock) showed a positive effect, whereas high glycerol concentration, tryptone and CaCl2 showed a negative impact on the 1,3-PDO produced by glycerol degradation. Following four experimental periods, the highest average yield of 0.43 mol 1,3-PDO mol-1 gly was achieved when sodium bicarbonate was added to the reactors. Propionate and acetate were also produced and a high microorganism diversity was detected; however, the restrictive operational conditions of the reactors led to the death of the methanogenic archaea. Nevertheless, the continuous production of 1,3-PDO from glycerol within UASB reactors inoculated with granular sludge can be considered highly feasible.

Multi-criteria analysis for municipal solid waste management in a Brazilian metropolitan area.

The decision-making process involved in municipal solid waste management (MSWM) must consider more than just financial aspects, which makes it a difficult task in developing countries. The Recife Metropolitan Region (RMR) in the Northeast of Brazil faces a MSWM problem that has been ongoing since the 1970s, with no common solution. In order to direct short-term solutions, three MSWM alternatives were outlined for the RMR, considering the current and future situations, the time and cost involved and social/environmental criteria. A multi-criteria approach, based on the Preference Ranking Organization Method for Enrichment Evaluations (PROMETHEE), was proposed to rank these alternatives. The alternative that included two private landfill sites and seven transfer, sorting and composting stations was confirmed as the most suitable and stable option for short-term MSWM, considering the two scenarios for the criteria weights. Sensitivity analysis was also performed to support the robustness of the results. The implementation of separate collections would minimize the amount of waste buried, while maximizing the useful life of landfill sites and increasing the timeframe of the alternative. Overall, the multi-criteria analysis was helpful and accurate during the alternative selection process, considering the similarities and restrictions of each option, which can lead to difficulties during the decision-making process.

Methanogenic activity inhibition by increasing the linear alkylbenzene sulfonate (LAS) concentration.

The effect of the initial concentration of linear alkylbenzene sulfonate (LAS) on specific methanogenic activity (SMA) was investigated in this work. Six anaerobic flasks reactors with 1 L of total volume were inoculated with anaerobic sludge (2 g VSS L(-1)). The reactors were assayed for 42 days, and fed with volatile fatty acids, nutrients, and LAS. The initial LAS concentrations were 0, 10, 30, 50, 75, and 100 mg L(-1) for the treatment flasks T1 (control), T2, T3, T4, T5, and T6, respectively. When compared with T1, T2 exhibited a 30% reduction in maximum SMA and total methane production (TMP). In treatment T3 through T6, the reductions were 44-97% (T3-T6) for SMA, and 30-90% (T3-T6) for TMP. Total LAS removal increased following the increase in the initial LAS concentration (from 36% at T1 to 76% at T6), primarily due to the high degree of sludge adsorption. LAS biodegradation also occurred (32% in all treatments), although this was most likely associated with the formation of non-methane intermediates. Greater removal by adsorption was observed in long-chain homologues, when compared to short-chain homologues (C13 > C10), whereas the opposite occurred for biodegradation (C10 > C13). The C13 homologue was adsorbed to a great extent (in mass) in T4, T5 and T6, and may also have inhibited methane formation in these treatments.

Partial nitrification and simultaneous denitrification in sequential anaerobic and aerobic reactors: performance and microbial community dynamics.

ABSTRACTThe removal of organic matter and nitrogen from domestic sewage was evaluated using a system composed of two sequential reactors: an anaerobic reactor (ANR) with suspended sludge and an aerobic (AER) reactor with suspended and adhered sludge to polyurethane foams. Nitrogen removal consisted of AER operating at low dissolved oxygen (DO) concentrations; this favoured the simultaneous nitrification and denitrification (SND) process. The concentration of COD and N were 440 mgO2.L-1 and 37 mgTN.L-1, respectively. The operation was divided into three phases (P), lasting 51, 53, and 46 days, respectively. The initial DO concentrations applied in the AER were: 3.0 (PI) and 1.5 mg.L-1 (PII and PIII). In PIII, the AER effluent was recirculated to the ANR at a ratio of 0.25. Kinetic assays were performed to determine the nitrification and denitrification rates of the biomasses (ANR and AER in PIII). Changes in the microbial community were evaluated throughout phases PI to PIII by massive sequencing. In PIII, the best results obtained for chemical oxygen demand (COD) and total nitrogen (TN-N) removal efficiencies, were close to 94% and 65%, respectively. Under these conditions, system effluent concentrations below 30 mg COD.L-1 and 15 mg TN-N.L-1 were verified. The nitritation and nitration rates were 10.5 and 6.5 mg N.g VSS-1.h-1, while the denitrification via nitrite and nitrate were 6.8 and 5.8 mg N.g VSS-1.h-1, respectively. A mixotrophic community was prevalent, with Rhodococcus, Nitrosomonas, Pseudomnas, and Porphyromonas being dominant or co-dominant in most of the samples, confirming the SND process in the AER sludge.

Bioremediation of a tropical clay soil contaminated with diesel oil.

The removal of polyaromatic hydrocarbons (PAH) in tropical clay soil contaminated with diesel oil was evaluated. Three bioremediation treatments were used: landfarming (LF), biostimulation (BS) and biostimulation with bioaugmentation (BSBA). The treatment removal efficiency for the total PAHs differed from the efficiencies for the removal of individual PAH compounds. In the case of total PAHs, the removal values obtained at the end of the 129-day experimental period were 87%, 89% and 87% for LF, BS and BSBA, respectively. Thus, the efficiency was not improved by the addition of nutrients and microorganisms. Typically, two distinct phases were observed. A higher removal rate occurred in the first 17 days (P-I) and a lower rate occurred in the last 112 days (P-II). In phase P-I, the zero-order kinetic parameter (µg PAH g(-1) soil d(-1)) values were similar (about 4.6) for all the three treatments. In P-II, values were also similar but much lower (about 0.14). P-I was characterized by a sharp pH decrease to less than 5.0 for the BS and BSBA treatments, while the pH remained near 6.5 for LF. Concerning the 16 individual priority PAH compounds, the results varied depending on the bioremediation treatment used and on the PAH species of interest. In general, compounds with fewer aromatic rings were better removed by BS or BSBA, while those with 4 or more rings were most effectively removed by LF. The biphasic removal behavior was observed only for some compounds. In the case of naphthalene, pyrene, chrysene, benzo[k]fluoranthene and benzo[a]pyrene, removal occurred mostly in the P-I phase. Therefore, the best degradation process for total or individual PAHs should be selected considering the target compounds and the local conditions, such as native microbiota and soil type.

Start-up strategies to develop aerobic granular sludge and photogranules in sequential batch reactors.

In this study, start-up strategies to develop conventional aerobic granular sludge (AGS) and algal aerobic granular sludge (AAGS) (photogranules), were investigated. The granulation experiment was conducted in four sequencing batch reactors (SBR), of which two were conventional SBRs (RC1, RC2) used as control, and two were photo-SBRs (R1, R2). R1 and RC1 were operated with a 40-min feeding during the reactors´ anaerobic cycle period, whereas R2 and RC2 with a 60-min feeding. All the reactors were operated in two phases with a C:N = 4:1 in Phase I and 8:1 in Phase II. In Phase I, AGS in RC1 and RC2 was formed 15 days before the AAGS development in R1 and R2. However, the AAGS generally presented better stability and higher diameter. On the other hand, AGS presented greater abundance of extracellular polymeric substances producing organisms, such as Xanthomonadacea and Rhodocyclaceae. Chemical oxygen demand (COD) and NH4+-N removal efficiencies were similar in all the four reactors of approximately 70% and 60%, respectively. In this phase, despite the good biomass structure, the reactors were not able to completely oxidize the high influent concentration of NH4+-N (100 mg.L-1) and COD (400 mg.L-1). This can be associated to the short time of the aerobic phase and low biomass content. In Phase II in all the reactors, a good increase in COD and NH4+-N removal efficiencies to values above 95% and 93%, respectively, was achieved under a higher C:N ratio of 8 with lower influent concentration of NH4+-N (50 mg.L-1). The 60-min anaerobic feeding period in R2 and RC2 resulted in greater removal efficiency of nitrogen, confirming that small variation on cycle periods can affect the biomass composition; the biomass presented more compact granules and larger diameters under 60 min-feeding when compared with those obtained with 40 min-feeding in Phase I.

Formation and stability of aerobic granular sludge in a sequential batch reactor for the simultaneous removal of organic matter and nutrients from low-strength domestic wastewater.

Simultaneous removal of organic matter, nitrogen, and phosphorus, via simultaneous nitrification and denitrification (SND) and enhanced biological phosphorus removal processes, was evaluated in a pilot-scale sequential batch reactor. The focus was on granule's morphology, stability, microbiological composition, and reactor performance while treating diluted domestic wastewater with total chemical oxygen demand (CODt) of ≈ 200 mg.L-1. The applied organic loading rate was 0.9 ± 0.3 kg CODt.m-3.d-1 in the experiment. Aerobic granular sludge developed gradually. After 87-day operation, granules (diameter ≥ 0.2 mm) were ≥ 50 % of the biomass, and after 168 days, complete granulation was obtained (≥ 80 % of biomass). In the third period (days 168-247, complete granulation), mixed liquor biomass reached a volatile suspended solids (VSS) concentration of 1.2 ± 0.3 g VSS.L-1, with the granules remaining stable until the experimental end. In this period, low effluent concentrations of COD, nitrogen (NH4+-N, NO2--N and NO3--N) and phosphate (PO43-P) were obtained (mg.L-1): 36 ± 11; 4 ± 5; 3 ± 3, 4 ± 5; and 0.9 ± 0.4, respectively. COD, NH4+-N, and PO43--P removal efficiencies (%) were 80 ± 11; 83 ± 20; and 55 ± 24, respectively. Heterotrophic nitrification and SND were observed, resulting in a process efficiency of 31 % even with dissolved oxygen applied to saturation. The phosphate removal was mainly attributed to denitrifying phosphorus accumulating organisms. Pseudomonas, the dominant genus found, acted in nitrogen and phosphorus removal. Pseudoxanthomonas also assisted in phosphorus removal. Bacterial communities in the flocs (≈ 20 % of biomass) during the last period were similar to those in the granules; therefore, they constituted the basis for granule formation, directly contributed to the simultaneous good removal of organic matter and nutrients.

The key role of oxygen in the bioremoval of 2,4-diaminoanisole (DAAN), the biotransformation product of the insensitive munitions compound 2,4-dinitroanisole (DNAN), over other electron acceptors.

Insensitive munitions compounds, such as 2,4-dinitroanisole (DNAN), are replacing conventional explosives. DNAN is anaerobically reduced to 2,4-diaminoanisole (DAAN), a toxic aromatic amine. However, the removal of DAAN under different redox conditions is yet to be elucidated. Herein, we analyzed DAAN consumption in biotic and abiotic microcosms when exposed to different redox conditions (without added electron acceptor, without added electron acceptor but with pyruvate as a co-substrate, with sulfate, with nitrate, and with oxygen), using an anaerobic sludge as inoculum. We observed that DAAN autoxidation, an abiotic reaction, was significant in microaerobic environments. DAAN also reacted abiotically with heat-killed sludge up to a saturation limit of 67.4 µmol DAAN (g VSS heat-killed sludge)-1. Oxygen caused the fastest removal of DAAN in live sludge among the conditions tested. Treatments without added electron acceptors (with or without pyruvate) presented similar DAAN removal performances, although slower than the treatment with oxygen. Sulfate did not exhibit any effect on DAAN removal compared to the treatment without added electron acceptors. Nitrate, however, inhibited the process. An enrichment culture from the microcosms exposed to oxygen could be developed using DAAN as the sole substrate in microaerobic conditions. The enrichment profoundly changed the microbial community. Unclassified microorganisms accounted for 85% of the relative abundance in the enrichment culture, suggesting that DAAN microaerobic removal might have involved organisms that were not yet described. Our results suggest that DAAN microaerobic treatment can be coupled to DNAN anaerobic reduction in sludge, improving the treatment of DNAN-containing wastewaters.

1,3-Propanediol production from glycerol in polyurethane foam containing anaerobic reactors: performance and biomass cultivation and retention.

The use of batch and upflow anaerobic reactors filled with polyurethane foam for pure glycerol fermentation was evaluated. The best reactor operational conditions to obtain high yield and productivity of 1,3-propanediol (1,3-PDO) as the main product and the role of the polyurethane foam in the growth and retention of suspended and attached biomass in the reactors were investigated. In the experiment at 30 °C with a batch reactor (700 mL), biomass growth was mostly as immobilized attached cells, and the achieved 1,3-PDO yield was up to 0.58 mol mol-gly-1. In the experiment (30 °C) with an upflow anaerobic reactor (717 mL), glycerol loading rates (gly-LR) ranging from 6.94 to 15.47 g gly L-1 day-1 were applied during a 102-day period. During the operation, average 1,3-PDO yield was 0.47 mol mol-gly-1, reaching a maximum of 0.51 mol mol-gly-1 at gly-LR of 13.57 g gly L-1 day-1. High 1,3-PDO productivity (5.35 to 5.44 g L-1 day-1) was obtained when gly-LR was 13.57 to 15.47 g gly L-1 day-1. Comparing the close yield values in both batch and continuous reactors and based on microbial evaluation, it is concluded that most of the 1,3-PDO generated in the continuous reactor was due to the suspended biomass retained by the foam cubes. The Clostridium genus was the predominant 1,3-PDO producer. Good yields and productivities with packed reactors were attributed to polyurethane foam used for mixed culture growth and retention. Consequently, they are worth considering for 1,3-PDO production from pure glycerol.

Evaluation of rhamnolipid addition on the natural attenuation of estuarine sediments contaminated with diesel oil.

The aim of the present study was to assess the bioremediation of estuarine sediments contaminated with diesel oil. The following two experiments were performed: natural attenuation (NA) and stimulated natural attenuation (SNA), using rhamnolipid as biosurfactant. Sediment samples were accommodated into glass columns and then contaminated with diesel oil on the top. The column profiles were separated into surface, middle, and bottom for the analyses. The 16 polycyclic aromatic hydrocarbons (PAHs) prioritized by US Environmental Protection Agency (EPA) were monitored for 349 days. Those with three and four rings showed increasing concentrations through the operation period in the middle and bottom samples, particularly between days 111 and 338, and in the SNA experiment. Those with five and six rings were also detected in the deeper portions of the columns, suggesting the percolation of PAHs with a high molecular weight. Total organic carbon was reduced by 91 and 89 % in the NA and SNA samples, respectively, although no statistically significant differences (p > 0.05) were found between the two treatments. The analyses by denaturing gradient gel electrophoresis indicated a slight shift in the microbial community structure over the experiments. Microorganisms belonging to the γ-Proteobacteria phylum were the main bacteria involved. The archaeal community exhibited dominance of hydrogenotrophic methanogens, indicating the obligate anaerobic biodegradation of intermediate compounds from hydrocarbon degradation.

Impact of pollution on the microbial diversity of a tropical river in an urbanized region of northeastern Brazil.

Rivers are important ecosystems that are integrated into biogeochemical cycles and constitute an essential resource for numerous human uses. However, the assessment of the biological diversity and composition of microbial communities found in rivers remains incomplete, partly due to methodological constraints which are only recently being resolved with the advent of next generation sequencing (NGS) techniques. Using 454-pyrosequencing of the 16S gene, the present study analyzed the microbial diversity of the planktonic and sediment populations in a tropical river in northeastern Brazil that is exposed to severe pollution. Six water and six sediment samples were analysed. The dominant bacterial phyla in both sediment and water were the Proteobacteria, followed by Bacteroidetes and Actinobacteria in the water column and by Chloroflexi and Acidobacteria in the sediment. Biological diversity appeared to be greatly decreased by environmental pollution, whereas the microbial community structure was variable across the analyzed transect. Moreover, a narrow relationship between industrial and urban sources of contamination and the bacterial genera detected at these sites has been observed. A variety of potentially pathogenic bacteria was detected, including Klebsiella, Treponema, Faecalibacterium and Enterococcus, indicating that the river might pose a substantial risk to public health. [Int Microbiol 20(1): 11-24 (2017)].

Natural attenuation and biosurfactant-stimulated bioremediation of estuarine sediments contaminated with diesel oil.

We evaluated the bioremediation, by natural attenuation (NA) and by natural attenuation stimulated (SNA) using a rhamnolipid biosurfactant, of estuarine sediments contaminated with diesel oil. Sediment samples (30 cm) were put into 35 cm glass columns, and the concentrations of the 16 polycyclic aromatic hydrocarbons (PAHs) prioritized by the US Environmental Protection Agency were monitored for 111 days. Naphthalene percolated through the columns more than the other PAHs, and, in general, the concentrations of the lower molecular weight PAHs, consisting of two and three aromatic rings, changed during the first 45 days of treatment, whereas the concentrations of the higher molecular weight PAHs, consisting of four, five, and six rings, were more stable. The higher molecular weight PAHs became more available after 45 days, in the deeper parts of the columns (20-30 cm). Evidence of degradation was observed only for some compounds, such as pyrene, with a total removal efficiency of 82 and 78 % in the NA and SNA treatments, respectively, but without significant difference. In the case of total PAH removal, the efficiencies were significantly different of 82 and 67 %, respectively.

Impact of pollution on the microbial diversity of a tropical river in an urbanized region of northeastern Brazil

Rivers are important ecosystems that are integrated into biogeochemical cycles and constitute an essential resource for numerous human uses. However, the assessment of the biological diversity and composition of microbial communities found in rivers remains incomplete, partly due to methodological constraints which are only recently being resolved with the advent of next generation sequencing (NGS) techniques. Using 454-pyrosequencing of the 16S gene, the present study analyzed the microbial diversity of the planktonic and sediment populations in a tropical river in northeastern Brazil that is exposed to severe pollution. Six water and six sediment samples were analysed. The dominant bacterial phyla in both sediment and water were the Proteobacteria, followed by Bacteroidetes and Actinobacteria in the water column and by Chloroflexi and Acidobacteria in the sediment. Biological diversity appeared to be greatly decreased by environmental pollution, whereas the microbial community structure was variable across the analyzed transect. Moreover, a narrow relationship between industrial and urban sources of contamination and the bacterial genera detected at these sites has been observed. A variety of potentially pathogenic bacteria was detected, including Klebsiella, Treponema, Faecalibacterium and Enterococcus, indicating that the river might pose a substantial risk to public health (AU)

No disponible