Treating protein-rich wastewater using cost-effective and simple-structured single-stage reactors presents several challenges. In this study, we applied an anaerobic sequencing batch reactor (AnSBR) to treat protein-rich wastewater from a slaughterhouse. We focused on identifying the key factors influencing the removal of chemical oxygen demand (COD) and the settling performance of the sludge. The AnSBR achieved a maximum total COD removal of 90%, a protein degradation efficiency exceeding 80%, and a COD to methane conversion efficiency of over 70% at organic loading rates of up to 6.2 g COD L⁻¹ d⁻¹. We found that the variations in both the organic loading rate within the reactor and the hydraulic retention time in the buffer tank had a significant effect on COD removal. The hydraulic retention time in the buffer tank and the reactor, which determined the ammonification efficiencies and the residual carbohydrate concentrations in the reactor liquid, affected the sludge settleability. Furthermore, the genus Clostridium sensu stricto 1, known as protein- and lipids-degraders, was predominant in the reactor. Statistical analysis showed a significant correlation between the core microbiome and ammonification efficiency, highlighting the importance of protein degradation as the governing process in the treatment. Our results will provide valuable insights to optimise the design and operation of AnSBR for efficient treatment of protein-rich wastewater.

Closing water loops in chemical industries result in hot and highly saline residual streams, often characterized by high strength and the presence of refractory or toxic compounds. These streams are attractive for anaerobic technologies, provided the chemical compounds are biodegradable. However, under such harsh conditions, effective biomass immobilization is difficult, limiting the use of the commonly applied sludge bed reactors. In this study, we assessed the long-term phenol conversion capacity of a lab-scale anaerobic membrane bioreactor (AnMBR) operated at 55°C, and high salinity (18 gNa^+.L^–1). Over 388 days, bioreactor performance and microbial community dynamics were monitored using specific methanogenic activity (SMA) assays, phenol conversion rate assays, volatile fatty acids permeate characterization and Illumina MiSeq analysis of 16S rRNA gene sequences. Phenol accumulation to concentrations exceeding 600 mgPh^.L^–1 in the reactor significantly reduced methanogenesis at different phases of operation, while applying a phenol volumetric loading rate of 0.12 gPh^.L^–1.d^–1. Stable AnMBR reactor performance could be attained by applying a sludge phenol loading rate of about 20 mgPh^.gVSS^–1.d^–1. In situ maximum phenol conversion rates of 21.3 mgPh^.gVSS^–1.d^–1 were achieved, whereas conversion rates of 32.8 mgPh^.gVSS^–1.d^–1 were assessed in ex situ batch tests at the end of the operation. The absence of caproate as intermediate inferred that the phenol conversion pathway likely occurred via carboxylation to benzoate. Strikingly, the hydrogenotrophic SMA of 0.34 gCOD-CH₄^.gVSS^–1.d^–1 of the AnMBR biomass significantly exceeded the acetotrophic SMA, which only reached 0.15 gCOD-CH₄^.gVSS^–1.d^–1. Our results indicated that during the course of the experiment, acetate conversion gradually changed from acetoclastic methanogenesis to acetate oxidation coupled to hydrogenotrophic methanogenesis. Correspondingly, hydrogenotrophic methanogens of the class Methanomicrobia, together with Synergistia, Thermotogae, and Clostridia classes, dominated the microbial community and were enriched during the three phases of operation, while the aceticlastic Methanosaeta species remarkably decreased. Our findings clearly showed that highly saline phenolic wastewaters could be satisfactorily treated in a thermophilic AnMBR and that the specific phenol conversion capacity was limiting the treatment process. The possibility of efficient chemical wastewater treatment under the challenging studied conditions would represent a major breakthrough for the widespread application of AnMBR technology.

This study examined the temperature susceptibility of a continuous-flow lab-scale anaerobic membrane bioreactor (AnMBR) to temperature shifts from 35 °C to 55 °C and its bioconversion robustness treating synthetic phenolic wastewater at 16 gNa^+.L⁻¹. During the experiment, the mesophilic reactor was subjected to stepwise temperature increases by 5 °C. The phenol conversion rates of the AnMBR decreased from 3.16 at 35 °C to 2.10 mgPh^.gVSS^−1.d⁻¹ at 45 °C, and further decreased to 1.63 mgPh^.gVSS^−1.d⁻¹ at 50 °C. At 55 °C, phenol conversion rate stabilized at 1.53 mgPh^.gVSS^−1.d⁻¹ whereas COD removal efficiency was 38% compared to 95.5% at 45 °C and 99.8% at 35 °C. Interestingly, it was found that the phenol degradation process was less susceptible for the upward temperature shifts than the methanogenic process. The temperature increase implied twenty-one operational taxonomic units from the reactor's microbial community with significant differential abundance between mesophilic and thermophilic operation, and eleven of them are known to be involved in aromatic compounds degradation. Reaching the upper-temperature limits for mesophilic operation was associated with the decrease in microbial abundance of the phyla Firmicutes and Proteobacteria, which are linked to syntrophic phenol degradation. It was also found that the particle size decreased from 89.4 μm at 35 °C to 21.0 μm at 55 °C. The accumulation of small particles and higher content of soluble microbial protein-like substances led to increased transmembrane pressure which negatively affected the filtration performance. Our findings indicated that at high salinity a mesophilic AnMBR can tolerate a temperature up to 45 °C without being limited in the phenol conversion capacity.