The anaerobic degradation of phenolic compounds presents substantial challenges due to their toxicity to methanogenic biomass and inherently low conversion rates. Recent studies indicate that nano-magnetite can stimulate direct interspecies electron transfer (DIET), potentially enhancing phenol conversion and methane production. This study employed two anaerobic membrane bioreactors (AnMBRs) to investigate phenol and p-cresol degradation under stepwise increasing loading rates, with complete retention of biomass in both reactors. While AnMBR-C served as a control, nano-magnetite was additionally supplemented to reactor AnMBR-M at a concentration of 40 mmol/L in Phase 1 and 20 mmol/L in Phase 2. Results demonstrated that AnMBR-M supplemented with 20 mmol/L nano-magnetite tolerated higher phenolic loading rates compared to AnMBR-C. In Phase 2, a higher total Fe concentration was observed in AnMBR-M, suggesting an enhanced electron transfer mechanism via dissimilatory iron reduction-oxidation cycle. Follow-up batch experiments revealed that magnetite-adapted biomass had more tolerance to phenol inhibition. A 16S-rRNA sequencing was conducted to characterize microbial communities within both reactors. Results suggested that DIET was stimulated in Phase 1, as shown by the enrichment of the electrogenic Pseudomonas and Methanolinea in AnMBR-M. However, the possibly stimulated DIET in Phase 1 could not alleviate the inhibition caused by excessive 40 mmol/L nano-magnetite dosage. Notably, there was no significant difference between the genera of AnMBR-C and AnMBR-M by the end of Phase 2. However, short-chain fatty acid degrader Mesotoga was more enriched in AnMBR-M. Moreover, species-level analysis showed that AnMBR-M had a sixfold higher relative abundance of Methanosaeta harundinacea compared to AnMBR-C.

Bitumen fume condensate (BFC) is a hazardous wastewater generated at asphalt reclamation and production sites. BFC contains a wide variety of potentially toxic organic pollutants that negatively affect anaerobic processes. In this study, we chemically characterized BFC produced at an industrial site and evaluated its degradation under anaerobic conditions. Analyses identified about 900 compounds including acetate, polycyclic aromatic hydrocarbons, phenolic compounds, and metal ions. We estimated the half maximal inhibitory concentrations (IC₅₀) of methanogenesis of 120, 224, and 990 mgCOD·L⁻¹ for three types of anaerobic biomass, which indicated the enrichment and adaptation potentials of methanogenic biomass to the wastewater constituents. We operated an AnMBR (7.0 L, 35 °C) for 188 days with a mixture of BFC, phenol, acetate, and nutrients. The reactor showed a maximum average COD removal efficiency of 87.7 ± 7.0 %, that corresponded to an organic conversion rate of 286 ± 71 mgCOD⁻¹·L⁻¹d⁻¹. The microbial characterization of the reactor's biomass showed the acetoclastic methanogen Methanosaeta as the most abundant microorganism (43 %), whereas the aromatic and phenol degrader Syntrophorhabdus was continuously present with abundances up to 11.5 %. The obtained results offer the possibility for the application of AnMBRs for the treatment of BFC or other petrochemical wastewater.

Phenol conversion under saline thermophilic anaerobic conditions requires the development and sustenance of a highly specialized microbial community. In the present research, an anaerobic membrane bioreactor (AnMBR) fed with an influent containing 0.5 g·L⁻¹ phenol and 6.5 gNa⁺·L⁻¹ was operated at 55 °C for 300 days. Phenol degradation was limited when phenol was the sole substrate. However, the phenol removal efficiency significantly (p < 0.001) increased to 80 % corresponding to a conversion rate of 29 mgPhenol·gVSS⁻¹d⁻¹ when acetate (0.5 gCOD·L⁻¹) was simultaneously provided. Isotopic analysis using 1–¹³C labeled acetate and measuring ¹³CH₄ revealed that acetate was first oxidized to hydrogen and CO₂, prior to methanogenesis, resulting in an increased abundance of hydrogenotrophic methanogens. It is hypothesized that the latter is of crucial importance for achieving effective anaerobic oxidation of phenol and its metabolites. Remarkably, the phenol conversion rate in the membrane-associated biomass was three times higher than in the suspended biomass. The observed difference in the conversion rate could be explained by the presence of an increased abundance of hydrogenotrophic methanogens in the membrane-associated biomass confirmed by a microbial community analysis of Archaea. Benzoate was measured in the permeate suggesting that phenol degradation occurred via the benzoyl-CoA pathway. The results of the current study suggest that syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis, which results in the presence of an abundant electron sink, plays a key role in enhancing thermophilic phenol degradation. The obtained insights widen the application of anaerobic digestion to treat saline phenolic-rich wastewater at high temperatures.

Treating petrochemical wastewater is a challenge for conventional anaerobic reactors. One example is coal gasification wastewater that, besides its salinity, is rich in toxic and inhibitory aromatics, such as phenol, cresols, and resorcinol. Studies have shown that phenol and p-cresol share the same degradation intermediates, whereas resorcinol is degraded via another route. This study investigated the simultaneous degradation of p-cresol or resorcinol with phenol under anaerobic saline conditions. Batch experiments with anaerobic phenol-degrading biomass were conducted to assess the feasibility of the degradation of p-cresol and resorcinol. Volumetric uptake rates of 11.4 ± 2.4 mg_p-cresol·L^–1d^–1 and 4.2 ± 1.9 mg_resorcinol·L^–1d^–1 were determined. The effect of p-cresol and resorcinol on the specific methanogenic activity and the cell viability in phenol-degrading and non-adapted biomass was assessed. Half maximal inhibitory concentration (IC₅₀) values of 0.73 g_p-cresol·L^-1 and 3.00 g_resorcinol·L^-1 were estimated for phenol-degrading biomass, whereas IC₅₀ values of 0.60 g_p-cresol·L^-1 and 0.25 g_resorcinol·L^-1 were estimated for the non-adapted biomass. p-Cresol caused a higher decrease in the non-damaged cell counts in comparison to resorcinol. Two anaerobic membrane bioreactors under saline conditions [8 g Na⁺·L^–1] were fed with mixtures of either phenol-p-cresol or phenol-resorcinol. At an influent phenol concentration of 2 g·L^-1, maximum conversion rates of 22 mg_p-cresol·gVSS^-1d^–1 and 16 mg_resorcinol·gVSS^–1d^–1 were found. In both AnMBRs, Syntrophorhabdus sp. and Methanosaeta sp. were the most abundant bacteria and methanogen, respectively. The feasibility of simultaneous conversion of phenolic compounds under saline conditions in AnMBRs opens novel perspectives for the high-rate anaerobic treatment of chemical wastewater.

Phenolic industrial wastewater, such as those from coal gasification, are considered a challenge for conventional anaerobic wastewater treatment systems because of its extreme characteristics such as presence of recalcitrant compounds, high toxicity, and salinity. However, anaerobic membrane bioreactors (AnMBRs) are considered of potential interest since they retain all micro-organism that are required for conversion of the complex organics. In this study, the degradation of phenol as main carbon and energy source (CES) in AnMBRs at high salinity (8.0 g Na⁺⋅L^–1) was evaluated, as well as the effect of acetate and an acetate-butyrate mixture as additional CES on the specific phenol conversion rate and microbial community structure. Three different experiments in two lab-scale (6.5 L) AnMBRs (35°C) were conducted. The first reactor (R1) was fed with phenol as the main CES, the second reactor was fed with phenol and either acetate [2 g COD⋅L^–1], or a 2:1 acetate-butyrate [2 g COD⋅L^–1] mixture as additional CES. Results showed that phenol conversion could not be sustained when phenol was the sole CES. In contrast, when the reactor was fed with acetate or an acetate-butyrate mixture, specific phenol conversion rates of 115 and 210 mgPh⋅gVSS^–1 d^–1, were found, respectively. The syntrophic phenol degrader Syntrophorhabdus sp. and the acetoclastic methanogen Methanosaeta sp. were the dominant bacteria and archaea, respectively, with corresponding relative abundances of up to 63 and 26%. The findings showed that dosage of additional CES allowed the development of a highly active phenol-degrading biomass, potentially improving the treatment of industrial and chemical wastewaters.