Degradation of long-chain fatty acids (LCFAs) in methanogenic environments is a syntrophic process involving the activity of LCFA-degrading bacteria and hydrogen-utilizing methanogens. If methanogens are inhibited, other hydrogen scavengers are needed to achieve complete LCFA degradation. In this work, we developed two different oleate (C_18:1 LCFA)-degrading anaerobic enrichment cultures, one methanogenic (ME) and another in which methanogenesis was inhibited (IE). Inhibition of methanogens was attained by adding a solution of 2-bromoethanesulfonate (BrES), which turned out to consist of a mixture of BrES and isethionate. Approximately 5 times faster oleate degradation was accomplished by the IE culture compared with the ME culture. A bacterium closely related to Syntrophomonas zehnderi (99% 16S rRNA gene identity) was the main oleate degrader in both enrichments, in syntrophic relationship with hydrogenotrophic methanogens from the genera Methanobacterium and Methanoculleus (in ME culture) or with a bacterium closely related to Desulfovibrio aminophilus (in IE culture). A Desulfovibrio species was isolated, and its ability to utilize hydrogen was confirmed. This bacterium converted isethionate to acetate and sulfide, with or without hydrogen as electron donor. This bacterium also utilized BrES but only after 3 months of incubation. Our study shows that syntrophic oleate degradation can be coupled to desulfonation. IMPORTANCE In anaerobic treatment of complex wastewater containing fat, oils, and grease, high long-chain fatty acid (LCFA) concentrations may inhibit microbial communities, particularly those of methanogens. Here, we investigated if anaerobic degradation of LCFAs can proceed when methanogens are inhibited and in the absence of typical external electron acceptors, such as nitrate, iron, or sulfate. Inhibition studies were performed with the methanogenic inhibitor 2-bromoethanesulfonate (BrES). We noticed that, after autoclaving, BrES underwent partial hydrolysis and turned out to be a mixture of two sulfonates (BrES and isethionate). We found out that LCFA conversion proceeded faster in the assays where methanogenesis was inhibited, and that it was dependent on the utilization of isethionate. In this study, we report LCFA degradation coupled to desulfonation. Our results also showed that BrES can be utilized by anaerobic bacteria.

The surfactant sodium lauryl ether sulfate (SLES) is widely used in the composition of detergents and frequently ends up in wastewater treatment plants (WWTPs). While aerobic SLES degradation is well studied, little is known about the fate of this compound in anoxic environments, such as denitrification tanks of WWTPs, nor about the bacteria involved in the anoxic biodegradation. Here, we used SLES as sole carbon and energy source, at concentrations ranging from 50 to 1000 mg L⁻¹, to enrich and isolate nitrate-reducing bacteria from activated sludge of a WWTP with the anaerobic-anoxic-oxic (A²/O) concept. In the 50 mg L⁻¹ enrichment, Comamonas (50%), Pseudomonas (24%), and Alicycliphilus (12%) were present at higher relative abundance, while Pseudomonas (53%) became dominant in the 1000 mg L⁻¹ enrichment. Aeromonas hydrophila strain S7, Pseudomonas stutzeri strain S8, and Pseudomonas nitroreducens strain S11 were isolated from the enriched cultures. Under denitrifying conditions, strains S8 and S11 degraded 500 mg L⁻¹ SLES in less than 1 day, while strain S7 required more than 6 days. Strains S8 and S11 also showed a remarkable resistance to SLES, being able to grow and reduce nitrate with SLES concentrations up to 40 g L⁻¹. Strain S11 turned out to be the best anoxic SLES degrader, degrading up to 41% of 500 mg L⁻¹. The comparison between SLES anoxic and oxic degradation by strain S11 revealed differences in SLES cleavage, degradation, and sulfate accumulation; both ester and ether cleavage were probably employed in SLES anoxic degradation by strain S11.

Mining and industrial activity are contributing to the increase in heavy metal (HM) pollution in soils. Phytoremediation coupled to selected rhizosphere microbiota is an environmentally friendly technology designed to promote HM bioremediation in soils. In this study, sunflower (Helianthus annuus L.) was used together with Rhizophagus irregularis, an arbuscular mycorrhizal fungi (AMF), and Cupriavidus sp. strain 1C2, a plant growth promoting rhizobacteria (PGPR), as a phytoremediation strategy to remove Zn and Cd from an industrial soil (599 mg Zn kg⁻¹ and 1.2 mg Cd kg⁻¹). The work aimed to understand if it is possible to gradually remediate the tested soil while simultaneously obtaining significant yields of biomass with further energetic values by comparison to the conventional growth of the plant in agricultural (non-contaminated) soil. The H. annuus biomass harvested in the contaminated industrial soil was 17% lower than that grown in the agricultural soil—corresponding to yields of 19, 620, 199 and 52 g m⁻² of roots, stems, flowers and seeds. It was possible to remove ca. 0.04 and 0.91% of the Zn and Cd of the industrial soil, respectively, via the HM accumulation on the biomass produced. The survival of applied microbiota was indicated by a high root colonization rate of AMF (about 50% more than in non-inoculated agricultural soil) and identification of strain 1C2 in the rhizosphere at the end of the phytoremediation assay. In this study, a phytoremediation strategy encompassing the application of an energetic crop inoculated with known beneficial microbiota applied to a real contaminated soil was successfully tested, with the production of plant biomass with the potential for upstream energetic valorisation purposes.