Hervé Macarie

Stenotrophomonas acidaminiphila sp. nov., a strictly aerobic bacterium isolated from an upflow anaerobic sludge blanket (UASB) reactor.

Two of several strictly aerobic, mesophilic bacteria isolated from a lab-scale upflow anaerobic sludge blanket (UASB) reactor treating a petrochemical wastewater, strains AMX 17 and AMX 19T, were subjected to detailed taxonomic study. Cells were gram-negative, motile, non-sporulating, straight to curved rods with a polar flagellum. The isolates exhibited phenotypic traits of members of the genus Stenotrophomonas, including cellular fatty acid composition and the limited range of substrates that could be used. Sugars and many amino acids were utilized. Antibiotic susceptibility and physiological characteristics were determined. The DNA base composition was 66.9 mol% G+C. Phylogenetic analysis revealed that the nearest relatives were Stenotrophomonas maltophilia LMG 11114, Stenotrophomonas nitritireducens DSM 12575T and Pseudomonas pictorum ATCC 23328T (similarity of 98.1-98.8%). Xanthomonas species, S. maltophilia LMG 958T and Stenotrophomonas africana CIP 104854T showed high 16S rRNA sequence similarities (96.4-97.3%). The high similarity found in cellular fatty acid profiles and identical partial 16S rRNA sequences (500 bp) for strains AMX 17 and AMX 19T indicate that they belong to the same species. DNA-DNA hybridizations revealed respectively 26.7, 31, 65.8 and 43.6% homology between isolate AMX 19T and S. africana CIP 104854T, S. maltophilia CIP 60.77T, S. nitritireducens DSM 12575T and P. pictorum ATCC 23328T. These results allow the proposal of strain AMX 19T (= DSM 13117T = ATCC 700916T = CIP 106456T) as representative of a novel species of the genus Stenotrophomonas, with the name Stenotrophomonas acidaminiphila sp. nov.

Multiple syntrophic interactions in a terephthalate-degrading methanogenic consortium

Terephthalate (TA) is one of the top 50 chemicals produced worldwide. Its production results in a TA-containing wastewater that is treated by anaerobic processes through a poorly understood methanogenic syntrophy. Using metagenomics, we characterized the methanogenic consortium inside a hyper-mesophilic (that is, between mesophilic and thermophilic), TA-degrading bioreactor. We identified genes belonging to dominant Pelotomaculum species presumably involved in TA degradation through decarboxylation, dearomatization, and modified β-oxidation to H2/CO2 and acetate. These intermediates are converted to CH4/CO2 by three novel hyper-mesophilic methanogens. Additional secondary syntrophic interactions were predicted in Thermotogae, Syntrophus and candidate phyla OP5 and WWE1 populations. The OP5 encodes genes capable of anaerobic autotrophic butyrate production and Thermotogae, Syntrophus and WWE1 have the genetic potential to oxidize butyrate to CO2/H2 and acetate. These observations suggest that the TA-degrading consortium consists of additional syntrophic interactions beyond the standard H2-producing syntroph–methanogen partnership that may serve to improve community stability.

Sporanaerobacter acetigenes gen. nov., sp. nov., a novel acetogenic, facultatively sulfur- reducing bacterium

A strictly anaerobic, moderately thermophilic, sporulating rod, designated strain Lup 33T, was isolated from an upflow anaerobic sludge blanket (UASB) reactor in Mexico. Strain Lup 33T possessed a few laterally inserted flagella, had a DNA G+C content of 32.2 mol % and grew optimally at pH 7.4 and 40 degrees C. Growth was observed at temperatures of up to 50 degrees C and was inhibited in the presence of 5% NaCl. Strain Lup 33T is heterotrophic and utilized some sugars, peptides and various single amino acids. Gelatin and casein were not used as energy sources. It performed the Stickland reaction and reduced elemental sulfur to sulfide. Acetate was the only fatty acid detected from glucose fermentation, whereas acetate together with isobutyrate and isovalerate were found as end products from peptone fermentation. Phylogenetically, strain Lup 33T branched with members of cluster XII of the order Clostridiales, with Clostridium hastiforme as the closest relative (similarity of 93%). On the basis of the phenotypic, genotypic and phylogenetic characteristics of the isolate, it is proposed as a novel species of a new genus, Sporanaerobacter acetigenes gen. nov., sp. nov. The type strain is strain Lup 33T (= DSM 13106T = CIP 106730T).

Clostridium thiosulfatireducens sp. nov., a proteolytic, thiosulfate- and sulfur-reducing bacterium isolated from an upflow anaerobic sludge blanket (UASB) reactor

A strictly anaerobic, gram-positive, sporulating rod (0.5-0.6 x 2.0-4.0 microm), designated strain Lup 21T, was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating cheese-factory wastewater. Strain Lup 21T was motile by means of peritrichous flagella, had a G+C content of 31.4 mol% and grew optimally at 37 degrees C, pH 7.4, in the absence of NaCl. It is a heterotrophic micro-organism, utilizing proteinaceous compounds (gelatin, peptides, Casamino acids and various single amino acids) but unable to use any of the carbohydrates tested as a carbon and energy source. It reduced thiosulfate and elemental sulfur to sulfide in the presence of Casamino acids as carbon and energy sources. Acetate, butyrate, isobutyrate, isovalerate, CO2 and sulfide were end products from oxidation of gelatin and Casamino acids in the presence of thiosulfate as an electron acceptor. In the absence of thiosulfate, serine, lysine, methionine and histidine were fermented. On the basis of 16S rRNA similarity, strain Lup 21T was related to members of the low-G+C Clostridiales group, Clostridium subterminale DSM 6970T being the closest relative (with a sequence similarity of 99.4%). DNA-DNA hybridization was 56% with this species. On the basis of phenotypic, genotypic and phylogenetic characteristics, the isolate was designated as a novel species of the genus Clostridium, Clostridium thiosulfatireducens sp. nov. The type strain is strain Lup 21T (= DSM 13105T = CIP 106908T).

Inhibition of the methanogenic fermentation of p-toluic acid (4-methylbenzoic acid) by acetate

The potential inhibitory effect of acetate on p-toluic acid methanogenic fermentation was studied during the continuous operation at 5.3 days hydraulic retention time of an upflow anaerobic sludge blanket reactor fed with a synthetic waste-water containing 3.67 mmp-toluic acid as sole carbon and energy source. In the absence of acetate, a chemical oxygen demand removal efficiency of 56.8% and an estimated p-toluic acid removal efficiency of 62.8% were achieved. Immediately after the addition of 58.3 mm acetate into the reactor influent, p-toluic acid degradation stopped while most of the acetate was consumed. The inhibition is explained by thermodynamic considerations. It is emphasized that such phenomena could occur during the treatment of waste-waters containing high concentrations of acetate and aromatic compounds that require a syntrophic association to be degraded to acetate and H2.

Anaerobic digestion Of a Petrochemical Wastewater using the UASB process

Anaerobic digestion of effluent from a petrochemical plant producing terephthalic acid has been tested using two Upflow Anaerobic Sludge Blanket (USAB) reactors. The reactors were seeded with two different inocula: one from an anaerobic stabilization pond receiving wasted sludge from the aerobic treatment plant of the petrochemical industry (reactor A); the other was an anaerobically adapted activated sludge from a municipal wastewater treatment plant (reactor B). At the beginning of the experiment, reactor A attained higher COD removal efficiencies and biogas production, but both reactors reached the same performances after 7 mo operation. The efficiencies in COD removal were low. At a 3 d hydraulic retention time (HRT), reactor A was loaded at 2.6 kg COD/m3 d and reactor B at 2.2 kg COD/m3 d. COD removals were 46.4 and 43.9% for reactor A and B, respectively. In view of these results, the UASB reactor does not appear as the most suitable treatment process for this kind of effluent.

Treatment of terephthalic acid plant wastewater with an anaerobic fixed film reactor

Abstract Anaerobic treatment of terephthalic acid plant wastewater with a lab‐scale downflow tubular fixed film reactor achieved 75% of COD removal with a HRT of 3.4 days at 33°C. The raw wastewater from a petrochemical plant was settled and neutralized prior to reactor feeding. The responses to shock loads and to periods without feeding were satisfactary, and no inhibitory effects were noticed. A primary settling ‐ anaerobic ‐ aerobic process configuration is proposed as a highly efficient, energy saving alternative to the conventional aerobic process.