(Per)chlorate reduction at high temperature: Physiological study of Archaeoglobus fulgidus and potential implications for novel souring mitigation strategies

The recent finding that Archaeoglobus fulgidus is able to couple (per)chlorate reduction to growth expanded this trait to the hyperthermophilic range of life. This sulfate-reducing archaeon is considered to be one of the major contributors to souring in hot oil reservoirs. Therefore, it is important to study its physiology in depth, particularly in view of novel souring mitigation strategies. A. fulgidus does not possess the classical (per)chlorate reduction pathway, as it lacks the key enzyme chlorite dismutase. Rather, the microorganism seems to couple (per)chlorate reduction to sulfur metabolism. Growth experiments show the strict necessity of sulfur compounds to sustain perchlorate reduction. Furthermore, the chemical formation of elemental sulfur was observed during perchlorate reduction, a compound that is biologically reduced again. Additional experiments showed that tetrathionate, but not elemental sulfur and polysulfide, serves as an electron acceptor for growth by A. fulgidus. Taken together these results provide further evidence for the importance of chemical and biological redox reactions involving sulfur compounds during (per)chlorate reduction. In non-reduced media also, nitrate could be reduced by A. fulgidus, though not coupled to growth. This observation and the fact that A. fulgidus had prolonged adaptation phases on sulfate after long-lasting growth on perchlorate are of interest in the development of new souring mitigation strategies using nitrate and/or (per)chlorate

Axenic mass cultivation of the free-living soil amoeba, Acanthamoeba castellanii in a laboratory fermenter

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Methanomethylovorans thermophila sp. nov., a thermophilic, methylotrophic methanogen form an anaerobic reactor fed with methanol

A novel thermophilic, obligately methylotrophic, methanogenic archaeon, strain L2FAWT, was isolated from a thermophilic laboratory-scale upflow anaerobic sludge blanket reactor fed with methanol as the carbon and energy source. Cells of strain L2FAWT were non-motile, irregular cocci, 0·7¿1·5 µm in diameter and usually occurred singly (sometimes forming clusters of two or four cells). The cells stained Gram-negative and lysed immediately in 0·1 % (w/v) SDS. Growth was inhibited by chloramphenicol and tetracycline, but not by penicillin, bacitracin, spectinomycin, vancomycin or kanamycin. Methanol and mono-, di- and trimethylamine were used as substrates, but H2/CO2, formate, acetate, propanol, dimethyl sulfide and methanethiol were not. The temperature range for growth was 42¿58 °C, with an optimum at 50 °C. The fastest growth was observed at a salinity below 100 mM NaCl; no growth occurred above 300 mM NaCl. The optimal pH for growth was 6·5; growth was observed from pH 5 to pH 7·5. The G+C content of the genomic DNA was 37·6 mol%. Analysis of the 16S rRNA gene sequence and the partial methyl-CoM reductase gene sequence revealed that the organism was phylogenetically closely related to Methanomethylovorans hollandica DMS1T (98 % similarity for the 16S rRNA gene sequence and 91 % similarity for the methyl-CoM reductase gene sequence). The DNA¿DNA relatedness between L2FAWT and Methanomethylovorans hollandica DMS1T was 46 %. On the basis of these results, strain L2FAWT (=DSM 17232T=ATCC BAA-1173T) represents the type strain of a novel species, for which the name Methanomethylovorans thermophila sp. nov. is proposed

Genome sequencing of one-carbon degrading acetogenic bacteria Moorella glycerini Strain NMP

The genus Moorella contains, up to now, only one sequenced genome, that of M. thermoacetica. Sequencing more strains is essential in understanding their common and specific physiology. Moreover, Moorella species typically grow with one-carbon substrates, but there are differences in their ability to grow with for example carbon monoxide. By comparing the already sequenced genomes of the type strains M. thermoacetica with close related strains we can assess which genes are responsible for the physiological differences

Volatile organic sulfur compounds in anaerobic sludge and sediments: biodegradation and toxicity

A variety of environmental samples was screened for anaerobic degradation of methanethiol, ethanethiol, propanethiol, dimethylsulfide, and dimethyldisulfide. All sludge and sediment samples degraded methanethiol, dimethylsulfide, and dimethyldisulfide anaerobically. In contrast, ethanethiol and propanethiol were not degraded by the samples investigated under any of the conditions tested. Methanethiol, dimethylsulfide, and dimethyldisulfide were mainly degraded by methanogenic archaea. In the presence of sulfate and the methanogenic inhibitor bromoethane sulfonate, degradation of these compounds coupled to sulfate reduction occurred as well, but at much lower rates. Besides their biodegradability, also the toxicity of methanethiol, ethanethiol, and propanethiol to methanogenesis with methanol, acetate, and H2/CO2 as the substrates was assessed. The 50% inhibition concentration of methanethiol on the methane production from these substrates ranged between 7 and 10 mM. The 50% inhibition concentration values of ethanethiol and propanethiol for the degradation of methanol and acetate were between 6 and 8 mM, whereas hydrogen consumers were less affected by ethanethiol and propanethiol, as indicated by their higher 50% inhibition concentration (14 mM). Sulfide inhibited methanethiol degradation already at relatively low concentrations: methanethiol degradation was almost completely inhibited at an initial sulfide concentration of 8 mM. These results define the operational limits of anaerobic technologies for the treatment of volatile organic sulfur compounds in sulfide-containing wastewater stream

Methanethiol degradation in anaerobic bioreactors at elevated pH (>8): Reactor performance and microbial community analysis

The degradation of methanethiol (MT) at 30 °C under saline¿alkaline (pH 8¿10, 0.5 M Na+) conditions was studied in a lab-scale Upflow Anaerobic Sludge Blanket (UASB) reactor inoculated with estuarine sediment from the Wadden Sea (The Netherlands). At a sodium concentration of 0.5 M and a pH between 8 and 9 complete MT degradation to sulfide, methane and carbon dioxide was possible at a maximum loading rate of 22 mmol MT L¿1 day¿1 and a hydraulic retention time of 6 h. The presence of yeast extract (100 mg/L) in the medium was essential for complete MT degradation. 16S rRNA based DGGE and sequence analysis revealed that species related to the genera Methanolobus and Methanosarcina dominated the archaeal community in the reactor sludge. Their relative abundance fluctuated in time, possibly as a result of the changing operational conditions in the reactor. The most dominant MT-degrading archaeon was enriched from the reactor and obtained in pure culture. This strain WR1, which was most closely related to Methanolobus taylorii, degraded MT, dimethyl sulfide (DMS), methanol and trimethylamine. Its optimal growth conditions were 0.2 M NaCl, 30 °C and pH 8.4. In batch and reactor experiments operated at pH 10, MT was not degrade