Anaerobic co-metabolic biodegradation of pharmaceuticals and personal care products driven by glycerol fermentation.

Anaerobic digestion in two sequential phases, acidogenesis and methanogenesis, has been shown to be beneficial for enhancing the biomethane generation from wastewater. In this work, the application of glycerol (GOH) as a fermentation co-substrate during the wastewater treatment was evaluated on the biodegradation of different pharmaceuticals and personal care products (PPCPs). GOH co-digestion during acidogenesis led to a significant increase in the biodegradation of acetaminophen (from 78 to 89%), ciprofloxacin (from 25 to 46%), naproxen (from 73 to 86%), diclofenac (from 36 to 48%), ibuprofen (from 65 to 88%), metoprolol (from 45 to 59%), methylparaben (from 64 to 78%) and propylparaben (from 68 to 74%). The heterotrophic co-metabolism of PPCPs driven by glycerol was confirmed by the biodegradation kinetics, in which kbio (biodegradation kinetics constant) values increased from 0.18 to 2.11 to 0.27-3.60 L g-1-VSS d-1, for the operational phases without and with GOH, respectively. The assessment of metabolic pathways in each phase revealed that the prevalence of aromatic compounds degradation, metabolism of xenobiotics by cytochrome P450, and benzoate degradation routes during acidogenesis are key factors for the enzymatic mechanisms linked to the PPCPs co-metabolism. The phase separation of anaerobic digestion was effective in the PPCPs biodegradation, and the co-fermentation of glycerol provided an increase in the generation potential of biomethane in the system (energetic potential of 5.0 and 6.3 kJ g-1-CODremoved, without and with GOH, respectively). This study showed evidence that glycerol co-fermentation can exert a synergistic effect on the PPCPs removal during anaerobic digestion mediated by heterotrophic co-metabolism.

Long-term exposure of Allonais inaequalis to a mixture of antibiotics in freshwater and synthetic wastewater matrices: Reproduction, recovery, and swimming responses.

Antibiotics from sulfonamide, fluoroquinolone, and diaminopyrimidine classes are widely used in human and veterinary medicine, and their combined occurrence in the aquatic environment is increasing around the world. In parallel, the understanding of how mixtures of these compounds affect non-target species from tropical freshwaters is scarce. Thus, this work aimed to study the long-term reproductive, recovery, and swimming effects of mixtures of 12 antibiotics from three different classes (up to 10 µg L-1 ) added to freshwater (FWM) and synthetic wastewater (SWM) matrices on freshwater worm Allonais inaequalis. Results revealed that at the reproduction level, the exposure to antibiotics in the SWM matrix does not cause a significant toxic effect on species after 10 days. On the other hand, exposures to initial dose mixtures (10 µg L-1 each) in FWM caused a significant reduction of offspring by 19.2%. In addition, recovery bioassays (10 days in an antibiotic-free environment) suggested that A. inaequalis has reduced offspring production due to previous exposure to antibiotic mixtures in both matrices. Furthermore, despite slight variation in swimming speed over treatments, no significant differences were pointed out. Regarding antibiotics in the water matrices after 10-day exposures, the highest concentrations were up to 2.7, 7.8, and 4.2 µg L-1 for antibiotics from sulfonamide, fluoroquinolone, and diaminopyrimidine classes, respectively. These findings suggest that a species positioned between primary producers and secondary consumers may experience late reproductive damage even in an antibiotic-free zone, after previous 10-day exposure to antibiotic mixtures. PRACTITIONER POINTS: A mixture of sulfonamide, fluoroquinolone, and diaminopyrimidine antibiotics in freshwater affects the offspring production of A. inaequalis after 10 days. After the 10-day antibiotic exposure, the reproduction of A. inaequalis remains affected in an antibiotic-free environment over the recovery period. The swimming speed of the worms does not change after 10 days of exposure to the antibiotic mixture. The concentration of dissolved solids can limit the natural degradation of sulfonamide, fluoroquinolone, and diaminopyrimidine antibiotics in the aquatic environment.

Understanding microbiome dynamics and functional responses during acidogenic fermentation of sucrose and sugarcane vinasse through metatranscriptomic analysis.

Improving anaerobic digestion of sugarcane vinasse - a high-strength wastewater from ethanol distillation - is a subject of great interest, in view of the reduction of the pollutants and recovery of methane and valuable metabolites as byproducts. Through metatranscriptomic analysis, this study evaluated the active microbiome and metabolic pathways in a continuous acidogenic reactor: Stage 1S (control): 100% sucrose-based substrate (SBS); Stage 2SV (acclimation): 50% SBS and 50% vinasse; Stage 3V: 100% vinasse. Metatranscriptome obtained from each Stage was subjected to taxonomic and functional annotations. Under SBS feeding, pH dropped to pH 2.7 and biohydrogen production was observed. As vinasse was added, pH increased to 4.1-4.5, resulting in community structure and metabolite changes. In Stage 3V, biohydrogen production ceased, and propionate and acetate prevailed among the volatile fatty acids. Release of homoacetogenesis enzymes by Clostridium ljungdahlii and of uptake hydrogenase (EC 1.12.99.6) by Pectinatus frisingensis were linked to hydrogen consumption in Stages 2SV and 3V. Metabolic pathways of vinasse compounds, such as carbohydrates, malate, oxalate, glycerol, sulfate and phenol, were investigated in detail. In pyruvate metabolism, gene transcripts of oadA (oxaloacetate decarboxylase) and mdh (malate dehydrogenase), were upregulated in Stage 3V, being mostly attributed to P. frisingensis. Acetate formation from vinasse degradation was mainly attributed to Megasphaera and Clostridium, and propionate formation to P. frisingensis. Glycerol removal from vinasse exceeded 99%, and gene transcripts encoding for glpF (glycerol uptake facilitator protein), glpK (glycerol kinase) and glpABC (glycerol-3-phosphate dehydrogenase) were expressed mostly by Pectinatus and Prevotella. mRNA profiling showed that active bacteria and gene expression greatly changed when vinasse replaced sucrose, and Pectinatus was the main active bacterium degrading the searched compounds from vinasse. The identification of the main metabolic routes and the associated microorganisms achieved in this work contributes with valuable information to support further optimization of fermentation towards the desired metabolites.

Improving the Continuous Multiple Tube Reactor: an Innovative Bioreactor Configuration with Great Potential for Dark Fermentation Processes.

The continuous multiple tube reactor (CMTR) has been developed as a promising technology to maximize biohydrogen production (BHP) by dark fermentation (DF) by preventing excess biomass accumulation, leading to suboptimum values of specific organic loading rates (SOLR). However, previous experiences failed to achieve stable and continuous BHP in this reactor, as the low biomass retention capacity in the tube region limited controlling the SOLR. This study goes beyond the evaluation of the CMTR for DF by inserting grooves in the inner wall of the tubes to ensure better cell attachment. The CMTR was monitored in 4 assays at 25ºC using sucrose-based synthetic effluent. The hydraulic retention time (HRT) was fixed at 2 h, while the COD varied between 2-8 g L-1 to obtain organic loading rates in the 24 - 96 g COD L-1 d-1. Long-term (90 d) BHP was successfully attained in all conditions due to the improved biomass retention capacity. Optimal values for the SOLR (4.9 g COD g-1 VSS d-1) were observed when applying up to 48 g COD L-1 d-1, in which BHP was maximized. These patterns indicate a favorable balance between biomass retention and washout was naturally achieved. The CMTR looks promising for continuous BHP and is exempt from additional biomass discharge strategies.

From start-up to maximum loading: An approach for methane production in upflow anaerobic sludge blanket reactor fed with the liquid fraction of fruit and vegetable waste.

This investigation provides a reproducible approach for determining the limits of an upflow anaerobic sludge blanket (UASB) reactor designed for the methanization of the liquid fraction of fruit and vegetable waste (FVWL). Two identical mesophilic UASB reactors were operated for 240 days with a three-day fixed hydraulic retention time and an organic load rate (OLR) increased from 1.8 to 10 gCOD L-1 d-1. Because of the previous estimation of flocculent-inoculum methanogenic activity, it was possible to design a safe OLR for the quick start-up of both UASB reactors. The operational variables obtained from the operation of the UASB reactors did not show statistical differences, ensuring the experiment's reproducibility. As a result, the reactors achieved methane yield close to 0.250 LCH4 gCOD-1 up to the OLR of 7.7 gCOD L-1 d-1. Furthermore, the maximum volumetric methane production rate of 2.0 LCH4 L-1 d-1 was discovered for the OLR ranges between 7.7 and 10 gCOD L-1 d-1. The possible overload at OLR of 10 gCOD L-1 d-1 resulted in a significant reduction of methane production in both UASB reactors. Based on the methanogenic activity of the UASB reactors sludge, a maximum loading capacity of approximately 8 gCOD L-1 d-1 was estimated.

Development of a low-cost electrochemical sensor for monitoring components in wastewater treatment processes.

Anaerobic digestion (AD) is a complex biological process widely used to decompose various types of organic matter, as well as to produce some metabolites and biogas. Diverse microorganism groups cooperate in many intricate metabolic routes so that organic matter can be degraded. However, any imbalance on these routes can lead to process instability or even failure. Therefore, a proper monitoring system, as well as a good understanding of the process, are key steps to improve performance and stability. Several mathematical models have been developed to represent AD. Despite this, process monitoring is mostly conducted by analytical methods, whose equipment is either expensive or the analyses are time-consuming, which may be a hindrance to low-budget developments. The objective of this study was to develop a low-cost electrochemical sensor to monitor components in wastewater treatment plants. Hundreds of synthetically supplemented sugarcane vinasse and synthetic domestic sewage samples were characterised. The obtained signals were used to calibrate principal component regression, partial least square and artificial neural network estimation models. The predictable variables were chemical oxygen demand, volatile fatty acids, sodium bicarbonate, beef extract, and lipids, and their R2 ranged from 0.84 to 0.99, depending on the component.

Automated detection and quantification of Enchytraeus crypticus (Oligochaeta: Enchytraeidae) in tropical artificial soil using image analysis.

The effects of toxic substance in soil matrices are evaluated by assessing adult worm survival and reproduction. Throughout the test, hundreds of juvenile potworms can be found. The current method for Enchytraeus crypticus quantification in soil samples is a laborious and time-consuming procedure that involves manual counting. The present work proposes a method for quick and reliable counting of E. crypticus by using an automated image analysis algorithm applied to soil images. Comparisons between automated and manual methods conducted in double-blind trials involving a large, routine batch of tropical artificial soil samples revealed no statistically significant differences for a wide range of worm densities. The proposed method overcomes time-consuming counts in manual methods and is suited to be deployed routinely for soil toxicity studies involving large batches of samples.

Proteogenomics identification of TBBPA degraders in anaerobic bioreactor.

Tetrabromobisphenol A (TBBPA) is the most used flame retardant worldwide and has become a threat to aquatic ecosystems. Previous research into the degradation of this micropollutant in anaerobic bioreactors has suggested several identities of putative TBBPA degraders. However, the organisms actively degrading TBBPA under in situ conditions have so far not been identified. Protein-stable isotope probing (protein-SIP) has become a cutting-edge technique in microbial ecology for enabling the link between identity and function under in situ conditions. Therefore, it was hypothesized that combining protein-based stable isotope probing with metagenomics could be used to identify and provide genomic insight into the TBBPA-degrading organisms. The identified 13C-labelled peptides were found to belong to organisms affiliated to Phytobacter, Clostridium, Sporolactobacillus, and Klebsilla genera. The functional classification of identified labelled peptides revealed that TBBPA is not only transformed by cometabolic reactions, but also assimilated into the biomass. By application of the proteogenomics with labelled micropollutants (protein-SIP) and metagenome-assembled genomes, it was possible to extend the current perspective of the diversity of TBBPA degraders in wastewater and predict putative TBBPA degradation pathways. The study provides a link to the active TBBPA degraders and which organisms to favor for optimized biodegradation.

Two-phase (acidogenic-methanogenic) anaerobic fixed bed biofilm reactor enhances the biological domestic sewage treatment: Perspectives for recovering bioenergy and value-added by-products.

The organic matter bioconversion into methane during anaerobic digestion (AD) comprises different steps, the acidogenic and methanogenic phases being clearly distinct in terms of metabolic activities. In this work, new configurations of anaerobic fixed bed biofilm reactors (AFBBR) were operated under conventional methanogenic conditions (single phase - SP-AFBBR, M1R), and in a sequential two-phase system, acidogenic reactor followed by methanogenic reactor (TP-AFBBR, AcR + M2R), in order to verify the impact of the AD phase separation on the overall system performance in operational, kinetics and microbiological aspects. The results indicated that feeding the methanogenic reactor with the acidogenic effluent stream provided a shorter operating start-up period (11 and 32 days for SP and TP-AFBBR, respectively), a greater alkalinity generation (0.14 and 0.41 g-CaCO3·g-CODremoved-1 for M1R and M2R, respectively), and the optimization of biomethane production (methane yield of 95 and 154 N-mLCH4·g-CODremoved-1 for M1R and M2R, respectively). The COD removal kinetics was also favored in the TP-AFBBR (k1-COD = 1.4 and 2.9 h-1 for M1R and M2R, respectively), since the soluble fermentation products were readily bioavailable to the biomass in the reactor. Hydrogenotrophic methanogenesis was the predominant pathway in the M2R, while the Methanosaeta-driven acetoclastic pathway predominated in the M1R. The greater diversity of Bacteria and Archaea in M2R denotes a better balance between the species that degrade volatile organic acids from AcR (i.e. Syntrophorhabdus, Syntrophus and Syntrophobacter) and the hydrogenotrophic methanogens (Methanoregula, Methanolinea and Methanospirillum) that consume the biodegradation products. The estimated bioenergy generation potential (range of 0.39-0.64 kWh·m-3-sewage considering the COD removed) for full-scale TP-sewage treatment plants evidences the feasibility of energetic recovery in the domestic sewage anaerobic treatment.

Selective removal and recovery of gallium and germanium from synthetic zinc refinery residues using biosorption and bioprecipitation.

The depletion of primary ores, the environmental concerns related to mining activities, and the need to promote circular economy has drawn attention to the recycling of metallic compounds. Bio-based technologies are suitable for metal recovery, as they operate under mild conditions (ambient temperature and pressure) and are ideal for treating low-concentration waters. This study compared the effectiveness of adsorption and precipitation for the removal and recovery of gallium, germanium and zinc. Adsorption of the metallic ions on elemental forms of sulfur (S0), selenium (Se0) and tellurium (Te0), both of chemical and biological sources, was tested. Biosorption onto elemental forms of S0bio, Se0bio and Te0bio effectively removed Ga and Zn. The highest removal efficiency (Õ²) was obtained for Ga onto the adsorbent Te0bio (69 ± 0.4%), with an adsorption capacity (q) of 74 mg Ga (g Te0bio)-1, followed by Zn (Õ² = 40 ± 0.7%) with 43 mg Zn (g Te0bio)-1. Precipitation with chemical and biogenic sulfide at different metal to sulfide (Me/S) ratios was also assessed. Biologically produced sulfide was more efficient for Ga and Zn compared to chemical sulfide. Precipitation with biogenic sulfide was efficient for the removal of Ga (Õ² = 59.9 ± 2.6%) and Zn (Õ² = 44.2 ± 3.0%). The lowest ratio between metal to sulfide (Me/S = 0.2) achieved higher zinc removal efficiencies, whereas gallium removal was more efficient at Me/S = 1.5. None of the tested methods allowed for recovery of Ge. Biosorption and bioprecipitation gave nevertheless high removal and recovery of Ga and Zn.

Hydrogen and organic acid production from dark fermentation of cheese whey without buffers under mesophilic condition.

BioH2 production from cheese whey (CW) was evaluated in two acidogenic reactors, UASB and structured fixed-bed (FB), without pH adjustment, under mesophilic conditions, and OLR of 25-90 g COD/L.d. Stage 1 was conducted as a control experiment using sucrose. BioH2 production occurred under pH < 3.0 with maximum yields of 5.8 and 3.0 mol H2/mol sucroseconsumed for UASB and FB reactors, respectively. In Stage 2, CW was the only substrate and a negligible bioH2 production was observed. Nevertheless, a maximum lactic acid concentration of 9.6 g/L was obtained, indicating that pH adjustment can be non-essential for lactic acid production from CW. In Stage 3, a strategy to enrich hydrogenogenic biomass was conducted by initially feeding the reactors with sucrose and gradually replacing it by CW. This strategy brought better bioH2 results compared to Stage 2, but it could not bear over the long-term, as non-hydrogen producing bacteria became predominant.

Tetrabromobisphenol A (TBBPA) biodegradation in acidogenic systems: One step further on where and who.

The occurrence of brominated flame retardants such as Tetrabromobisphenol A (TBBPA) in water bodies poses a serious threat to aquatic ecosystems. Degradation of TBBPA in wastewater has successfully been demonstrated to occur through anaerobic digestion (AD), although the involved microorganisms and the conditions favouring the conversion remains unclear. In this study, it was observed that bioconversion of TBBPA did not occur during the hydrolytic stage of the AD, but during the strictly fermentative stage. Bioconversion occurred in hydrolytic-acidogenic as well as in strictly acidogenic continuous bioreactors. This indicates that the microorganisms that degrade TBBPA benefit from the electron flux taking place during glycolysis and further transformations into short-chain fatty acids. The degradation kinetics of TBBPA was inversely proportional to the complexity of the wastewater as the apparent kinetics constants were 2.11, 1.86, and 0.52 h-1·gVSS-1 for glucose, starch, and domestic sewage as carbon source, respectively. Additionally, the micropollutant loading rate relative to the overall organic loading rate is of major importance during the investigation of cometabolic transformations. The long-term exposure to TBBPA at environmentally realistic concentrations did not cause any major changes in the microbiome composition. Multivariate statistical analysis of the evolvement of the microbiome throughout the incubation suggested that Enterobacter spp. and Clostridium spp. are the key players in TBBPA degradation. Finally, a batch enrichment was conducted, which showed that concentrations of 0.5 mg·L-1 or higher are detrimental to Clostridium spp., even though these organisms are putative TBBPA degraders. The Clostridium genus was outcompeted by the Enterobacter and Klebsiella genera, hereby highlighting the effect of unrealistic concentrations frequently used in culture-dependent studies on the microbial community composition.

Counting Enchytraeus crypticus Juveniles in Chronic Exposures: An Alternative Method for Ecotoxicity Studies Using Tropical Artificial Soil.

Soil toxicity tests are commonly applied using Enchytraeus crypticus to analyze reproductive outputs. However, the traditional method for counting potworms takes a long time due to the significant number of offspring. This paper compares the conventional total counting of E. crypticus juveniles (M1) and an alternative methodology (M2). The proposed methodology (M2) uses a simple random counting method (1/4) for the partial counting of juveniles and total estimation. Chronic bioassays (21 days of exposure) were performed in tropical artificial soil (TAS) using sugarcane vinasse as a hazardous substance. Comparing the final density of juveniles recorded in M1 and M2, no statistical differences were pointed out in either one. Applying analyses based on effective concentration (EC10 and EC50), no statistical differences were identified there either. The t-test showed that there was no statistical difference between the counting methods (M1 and M2) in each treatment (control and dilutions). Moreover, we ran the Tukey test for M1 and M2 methods separately and observed that 100 % of the vinasse showed a statistical difference compared to the control treatment in both (p ≤ 0.05), affirming that independent of the counting method, the ecotoxicological outputs were similar. Therefore, the proposed alternative is a suitable method for bioassay using. E. crypticus in tropical artificial soil, decreasing to 1/4 the total time required for counting.

Evaluation of the influence of trace metals on methane production from domestic sewage, using the Plackett-Burman experimental design.

Methanogenesis involves several enzymes with trace metal components that catalyze major metabolic pathways and, therefore, requires a sufficient supply of micronutrients such as iron, nickel or cobalt. The statistically-based Plackett-Burman experimental design was adopted in this study to identify which trace metal have a statistically significant effect on the maximum methane production from domestic sewage. The addition of Barium (Ba), Cobalt (Co), Copper (Cu), Iron (Fe), Manganese (Mn), Nickel (Ni) and Selenium (Se) was tested in batch reactors using domestic sewage as the substrate and sewage sludge as the inoculum. The results showed that the addition of Fe was statistically significant, positively affecting the maximum CH4 production (p-value 0.05). The results are expressed in L of CH4 per g of Chemical Oxygen Demand added, increasing it from 0.10 to 0.13 LCH4 gCODadd-1. In L of CH4 per g of Volatile Solids, increased it from 0.031 to 0.040 LCH4 gVS-1. The addition of Se was statistically significant, but with a negative effect on the maximum methane production (p-value 0.002), which decreased from 0.13 to 0.10 LCH4.gCODadd-1. Moreover, six different Fe concentrations (0, 40, 80, 120, 160 and 200 mgL-1) were tested, showing that the addition of 120 mgL-1 had the greatest effect for statistically improving the maximum methane production, with 33% improvement (0.12 ± 0.003 to 0.16 ± 0.012 LCH4 gCODadd-1) compared to no addition of Fe and the specific CH4 production to 0.040 ± 0.001 LCH4 gVS-1.

Tetrabromobisphenol A (TBBPA) anaerobic biodegradation occurs during acidogenesis.

This is the first study to bring evidence on the anaerobic biodegradation of TBBPA occurring during acidogenesis in domestic sewage at environmentally relevant concentrations by complex microbial communities. This was accomplished by continuously operating two anaerobic structured bed reactors (ASTBR) for over 100 days under acidogenic (Acidogenic Reactor, AR) and multistep methanogenic (Methanogenic Reactor, MR) conditions. In the AR, the temporal carbohydrates consumption and the acetic acid production were strongly correlated with TBBPA removal by the Pearson's test. The spatial concentration of TBBPA and carbohydrates along the MR and the kinetic degradation profiles corroborate the AR results. It is hypothesized that TBBPA biodegradation in the studied conditions occurs during acidogenesis via the cometabolism supported by non-specific enzymes and the metabolism (dehalorespiration) established by electrons donors such as H2, which are both produced during the macrocomponents breakdown in the early stages of the anaerobic digestion. The TBBPA mass balance showed that approximately 86.8 ± 0.05% and 97 ± 0.01% of the removed TBBPA was biodegraded in the AR and MR, respectively. Furthermore, TBBPA biodegradation went further than reductive debromination as total phenols were detected in the reactors' effluent.

Sugarcane vinasse extreme thermophilic digestion: a glimpse on biogas free management.

The high temperature in which sugarcane vinasse (SV) is generated (~ 90 °C) and the positive effect of higher temperatures in biochemical reactions have motivated the evaluation of SV anaerobic digestion (AD) under extreme temperature conditions. Two-stage (acidogenic/methanogenic) and single-stage (methanogenic) AD of SV were evaluated under 70 °C in structured-bed reactors. The extreme temperature was beneficial to the acidogenic step of the two-stage AD process. The methane production, however, was hindered at 70 °C. The VMP of the single and two-stage reactors accounted, respectively, for only 13% and 7% of the production rate reported in sugarcane vinasse AD at 55 °C. At 70 °C, the main genera responsible for methane production was Methanothermobacter and the acetoclastic methanogenesis did not occur, resulting in acetic acid build up (15,800 mg L-1). These results brought a new perspective for sugarcane vinasse management, with acetic acid production alternatively to methanization. In this perspective, two-stage process would be composed of acidogenic and acetogenic reactors, and beyond acetate, hydrogen and other soluble compounds could be recovered in a complete biorefinery process.

Modeling anaerobic digestion metabolic pathways for antibiotic-contaminated wastewater treatment.

Anaerobic digestion has been used to treat antibiotic-contaminated wastewaters. However, it is not always effective, since biodegradation is the main removal mechanism and depends on the compound chemical characteristics and on how microbial metabolic pathways are affected by the reactor operational conditions and hydrodynamic characteristics. The aim of this study was to develop a mathematical model to describe 16 metabolic pathways of an anaerobic process treating sulfamethazine-contaminated wastewater. Contois kinetics and a useful reaction volume term were used to represent the biomass concentration impact on bed porosity in a N continuously stirred tank modeling approach. Two sulfamethazine removal hypotheses were evaluated: an apparent enzymatic reaction and a cometabolic degradation. Additionally, long-term modeling was developed to describe how the operational conditions affected the performance of the process. The best degradation correlations were associated with the consumption of carbohydrates, proteins and it was inversely related to acetic acid production during acidogenesis.

Micro-Oxygenation in Upflow Anaerobic Sludge Bed (UASB) Reactors Using a Silicon Membrane for Sulfide Oxidation.

Sulfide produced by sulphate-reducing bacteria in anaerobic reactors can seriously affect biogas quality. Microaeration has become a reliable way to remove sulfide, by promoting its oxidation. However, limited research is available regarding its application in upflow anaerobic sludge bed (UASB) reactors. In this research, silicon membranes were studied as a mechanism to dose oxygen in USAB reactors. Two configurations were tested: the membrane placed inside the reactor or in an external module. Our results show that the external membrane proved to be a more practical alternative, providing conditions for sulfide oxidation. This led to a reduction in its concentration in the liquid effluent and biogas. External membrane configuration achieved a sulfide conversion rate of 2.4 g-S m2 d-1. Since the membrane was not sulfide-selective, methane losses were observed (about 9%). In addition, excessive oxygen consumption was observed, compared to the stoichiometric requirement. As is the case for many membrane-based systems, membrane area is a key factor determining the correct operation of the system.

Application of Dispersive Liquid-Liquid Microextraction Followed by High-Performance Liquid Chromatography/Tandem Mass Spectrometry Analysis to Determine Tetrabromobisphenol A in Complex Matrices.

An accurate and sensitive ultrasound-dispersive liquid-liquid microextraction technique followed by high-performance liquid chromatography separation coupled with electrospray ionization tandem mass spectrometry detection method to determine the presence of tetrabromobisphenol A (TBBPA) in complex environmental matrices is proposed. The miniaturized procedure was used to extract and quantify the analyte in domestic sewage, anaerobic sludge, and the aquatic test organism species Daphnia magna and Chironomus sancticaroli, which are standardized organisms for ecotoxicity bioassays. Limits of detection of 2 ng L-1 (domestic sewage), 2 ng g-1 (anaerobic sludge), 0.25 ng g-1 (D. magna), and 5 ng g-1 (C. tentans) were obtained. The presence of TBBPA was determined in domestic sewage and anaerobic sludge from an anaerobic batch bioreactor at a concentration of 0.2 ± 0.03 µg L-1 and 507 ± 79 ng g-1 , respectively. In D. magna and C. sancticaroli exposed to TBBPA in an acute toxicity bioassay, the micropollutant accumulated at 3.74 and 8.87 µg g-1 , respectively. The proposed method is a simple and cost-effective tool to determine TBBPA environmental occurrence and biomagnification potential compared with conventional extraction methods. To the best of our knowledge, this is the first liquid-liquid miniaturized extraction method to be applied to D. magna and C. sancticaroli. Environ Toxicol Chem 2020;39:2147-2157. © 2020 SETAC.

Influence of organic loading rate on ciprofloxacin and sulfamethoxazole biodegradation in anaerobic fixed bed biofilm reactors.

Antibiotic compounds, notably sulfamethoxazole (SMX) and ciprofloxacin (CIP), are ubiquitous emerging contaminants (ECs), which are often found in domestic sewage. They are associated with the development of antimicrobial resistance. Operational parameters, e.g. organic loading rate (OLR), hydraulic retention time (HRT) and sludge retention time, may influence EC biodegradation in wastewater treatment plants. This study assessed the impact of the OLR variation on the biodegradation of CIP and SMX, applying two configurations of anaerobic fixed bed reactors: anaerobic packed bed biofilm reactor (APBBR) and anaerobic structured bed biofilm reactor (ASBBR). A significant reduction in the biodegradation of SMX (APBBR: 93-69%; ASBBR: 94-81%) and CIP (APBBR: 85-66%; ASBBR: 85-64%) was observed increasing OLR from 0.6 to 2.0 kgCOD m-3 d-1. The decrease in the HRT from 12 to 4 h resulted in higher liquid-phase mass transfer coefficient (APBBR: ks from 0.01 to 0.05 cm h-1; ASBBR: ks from 0.07 to 0.24 cm h-1), but this was not enough to overcome the decrease in the antibiotic-biomass contact time on biofilm, thus reducing the bioreactors' performance. The ASBBR favored biomethane production (from 7 to 17 mLCH4 g-1VSS L-1 d-1) and biodegradation kinetics (kbio from 1.7 to 4.2 and for SMX and from 2.1 to 4.8 L g-1VSS d-1 for CIP) due to the higher relative abundance of the archaea community in the biofilm and the lower liquid-phase mass transfer resistance in the structured bed. CIP and SMX cometabolic biodegradation was associated to the hydrogenotrophic methanogenesis (mainly Methanobacterium genus) in co-culture with fermentative bacteria (notably the genera Clostridium, Bacillus, Lactivibrio, Syntrophobacter and Syntrophorhabdus). The anaerobic fixed bed biofilm reactors proved to be highly efficient in biodegrading the antibiotics, preventing them from spreading to the environment.