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International audience; Anaerobic digestion makes it possible to valorize municipal solid waste (MSW) into biogas and digestate which are, respectively, a renewable energy source and an organic amendment for soil. Phenols are persistent pollutants present in MSW that can inhibit the anaerobic digestion process and have a toxic effect on microbiota if they are applied to soil together with digestate. It is then important to define the operational conditions of anaerobic digestion which allow the complete degradation of phenol. In this context, the fate of phenol during the anaerobic digestion of MSW at 55 °C was followed using an isotopic tracing approach (13C6-phenol) in experimental microcosms with inoculum from an industrial thermophilic anaerobic digester. With this approach, it was possible to demonstrate the complete phenol biodegradation into methane and carbon dioxide via benzoate. Benzoate is known to be a phenol metabolite under mesophilic conditions, but in this study it was found for the first time to be a phenol degradation product at thermophilic temperature.

During this thesis, I studied the early evolution of life, from the Last Universal Common Ancestor (LUCA) to the ancestors of the three kingdoms, Archaea, Bacteria and Eukarya. Notably, I have attempted to place a few organisms in the tree of life, namely the bacteria Aquifex aeolicus and the archaea Cenarchaeum symbiosum, and I also studied the evolution of optimal growth temperatures over the last four billion years. To this end, I developed algorithms to reconstruct ancestral gene sequences, and used these sequences to predict the optimal growth temperatures of now-extinct organisms. My colleagues and I estimate that LUCA did not live in a very hot environment, but that its descendants the ancestors of Bacteria and of the group containing Archaea and Eukarya both lived at higher temperatures. This implies that the two lineages descending from LUCA underwent the same kind of evolution in parallel, perhaps caused by the same unique selection pressure. This pressure may have resulted from an intense meteoritic bombardment 3.8 billion years ago, and have been accompanied by the transition from an RNA genome in LUCA to DNA genomes in its descendants. Subsequently in the bacterial lineage, optimal growth temperature dropped, which may correspond to the evolution of oceanic temperatures in the last 3.5 billion years.; Durant cette thèse je me suis intéressé à l'évolution profonde du vivant, depuis le dernier ancêtre commun universel (LUCA) jusqu'aux ancêtres des trois grands royaumes, les Archées, les Bactéries et les Eucaryotes. J'ai notamment cherché à placer quelques organismes dans l'arbre de la vie, tels que la bactérie Aquifex aeolicus et l'archée Cenarchaeum symbiosum, et j'ai également étudié l'évolution des températures de croissance il y a plusieurs milliards d'années. Pour ce faire, j'ai développé des algorithmes afin de reconstruire l'évolution de séquences géniques, puis j'ai utilisé ces séquences pour prédire les températures optimales de croissances d'organismes aujourd'hui éteints. Mes collègues et moi-même estimons que LUCA ne vivait pas à très haute température, mais que ses directs descendants les ancêtres des Bactéries et du groupe comprenant les Archées et les Eucaryotes vivaient dans des environnements plus chauds. Cela signifie que les deux lignées venant de LUCA ont subi le même type d'évolution en parallèle, qui pourrait avoir été causée par une seule et même pression de sélection. Cette pression pourrait être le résultat d'un intense bombardement météoritique il y a 3.8 milliards d'années, et avoir été accompagnée d'un changement depuis un génome à ARN pour LUCA vers des génomes à ADN pour ses descendants. Ensuite, dans la lignée des Bactéries, les températures optimales de croissance ont chuté, ce qui pourrait correspondre à l'évolution de la température des océans au cours des 3.5 derniers milliards d'années.

During this thesis, I studied the early evolution of life, from the Last Universal Common Ancestor (LUCA) to the ancestors of the three kingdoms, Archaea, Bacteria and Eukarya. Notably, I have attempted to place a few organisms in the tree of life, namely the bacteria Aquifex aeolicus and the archaea Cenarchaeum symbiosum, and I also studied the evolution of optimal growth temperatures over the last four billion years. To this end, I developed algorithms to reconstruct ancestral gene sequences, and used these sequences to predict the optimal growth temperatures of now-extinct organisms. My colleagues and I estimate that LUCA did not live in a very hot environment, but that its descendants the ancestors of Bacteria and of the group containing Archaea and Eukarya both lived at higher temperatures. This implies that the two lineages descending from LUCA underwent the same kind of evolution in parallel, perhaps caused by the same unique selection pressure. This pressure may have resulted from an intense meteoritic bombardment 3.8 billion years ago, and have been accompanied by the transition from an RNA genome in LUCA to DNA genomes in its descendants. Subsequently in the bacterial lineage, optimal growth temperature dropped, which may correspond to the evolution of oceanic temperatures in the last 3.5 billion years.; Durant cette thèse je me suis intéressé à l'évolution profonde du vivant, depuis le dernier ancêtre commun universel (LUCA) jusqu'aux ancêtres des trois grands royaumes, les Archées, les Bactéries et les Eucaryotes. J'ai notamment cherché à placer quelques organismes dans l'arbre de la vie, tels que la bactérie Aquifex aeolicus et l'archée Cenarchaeum symbiosum, et j'ai également étudié l'évolution des températures de croissance il y a plusieurs milliards d'années. Pour ce faire, j'ai développé des algorithmes afin de reconstruire l'évolution de séquences géniques, puis j'ai utilisé ces séquences pour prédire les températures optimales de croissances d'organismes aujourd'hui éteints. Mes collègues et moi-même estimons que LUCA ne vivait pas à très haute température, mais que ses directs descendants les ancêtres des Bactéries et du groupe comprenant les Archées et les Eucaryotes vivaient dans des environnements plus chauds. Cela signifie que les deux lignées venant de LUCA ont subi le même type d'évolution en parallèle, qui pourrait avoir été causée par une seule et même pression de sélection. Cette pression pourrait être le résultat d'un intense bombardement météoritique il y a 3.8 milliards d'années, et avoir été accompagnée d'un changement depuis un génome à ARN pour LUCA vers des génomes à ADN pour ses descendants. Ensuite, dans la lignée des Bactéries, les températures optimales de croissance ont chuté, ce qui pourrait correspondre à l'évolution de la température des océans au cours des 3.5 derniers milliards d'années.

Thermophilic degradation of pectin was studied in batch cultures at 55°C by different associations of anaerobic bacteria, includingClostridium thermocellum, Methanobacterium sp., andMethanosarcina sp.Clostridium thermocellum alone produced large amounts of methanol along with some isopropanol and H2. The inoculation ofMethanobacterium sp. in the culture did not affect the metabolism ofC. thermocellum; this demonstrates the absence of interspecies hydrogen transfer. In the presence of the methylotrophicMethanosarcina sp., methanol was reduced to methane without effect on pectin hydrolysis; a small amount of the H2 produced was also used to reduce methanol.

Forty-four fungal strains capable of growing at temperatures above 50 C were isolated from different samples of soil and coconut residues collected in Mexican tropical and subtropical regions. These thermophilic and thermotolerant fungal strains were identified by microscopical analysis using standard procedures. Three species were identified: Rhizomucor pusillus (Lindt)Schipper (19 strains), Rhizopus microsporus van Tieghem (6 strains), and Aspergillus fumigatus Fresenius (19 strains). Four strains identified as Rhizopus microsporus were ascribed to the variety rhizopodiformis; however, the other two strains showed new characteristics which require further analysis, such as a homothallic sexual reproduction. Aspergillus fumigatus was found in coconut residues as a common contaminant during the isolation of other thermophilic species. Strains were isolated from samples containing a high content of lipids (mainly from coconut coprah), and accordingly extracellular lipase biosynthesis was directly confirmed in Petri dishes for every strain.

International audience; Le 26 décembre 1999, la tempête qui venait de traverser la France s'est abattue sur la Lorraine. Elle a durement touché l'arboretum d'Amance, causant des dégâts importants mais épargnant la majeure partie des collections. Un premier bilan des destructions a été établi : il montre que 3,6 % des espèces feuillues et 5,7 % des espèces résineuses ont disparu. D'autres espèces sont menacées de disparition : celles dont il ne reste plus qu'un ou deux exemplaires, parfois en mauvais état. C'est le cas pour 5 % des espèces feuillues restantes et pour 12,9 % des espèces résineuses restantes. C'est le secteur "Amérique de l'Ouest, côte Pacifique" qui a été le plus durement touché, suivi du secteur "Amérique de l'Est, côte Atlantique". L'arboretum de collection d'Amance n'est pas détruit, et sa remise en état, grâce aux subsides exceptionnels qui ont déjà été dégagés par l'INRA, est bien avancée. Elle se poursuivra par la remise en état des parcelles et par l'introduction (ou la réintroduction) d'espèces plus thermophiles, pour prendre en compte le changement climatique que nous sommes probablement en train de vivre

This article is free to read on the publishers website Several members of the order Thermotogales in the domain Bacteria, viz., Thermotoga neapolitana, Thermotoga maritima, Thermosipho africanus, Fervidobacterium islandicum, and Thermotoga strain SEBR 2665, an isolate from an oil well, reduced thiosulfate to sulfide. This reductive process enhanced cellular yields and growth rates of all the members but was more significant with the two hyperthermophiles T. neapolitana and T. maritima. This is the first report of such an occurrence in this group of thermophilic and hyperthermophilic anaerobic bacteria. The results suggest that thiosulfate reduction is important in the geochemical cycling of sulfur in anaerobic thermal environments such as the slightly acidic and neutral-pH volcanic hot springs and oil reservoirs.

Strain MLFT (T = type strain), a new thermophilic, spore-forming sulfate-reducing bacterium, was characterized and was found to be phenotypically, genotypically, and phylogenetically related to the genus Desulfotomaculum. This organism was isolated from a butyrate enrichment culture that had been inoculated with a mixed compost containing rice hulls and peanut shells. The optimum temperature for growth was 50 degrees C. The G+C content of the DNA was 51.2 mol%. Strain MLFT incompletely oxidized pyruvate, butyrate, and butanol to acetate and presumably CO2. It used long-chain fatty acids and propanediols. We observed phenotypic and phylogenetic differences between strain MLFT and other thermophilic Desulfotomaculum species that also oxidize long-chain fatty acids. On the basis of our results, we propose that strain MLFT is a member of a new species, Desulfotomaculum thermosapovorans.

A thermophilic, glucose-fermenting, strictly anaerobic, rod-shaped bacterium, strain SEBR 6459T (T = type strain), was isolated from an African oil-producing well. This organism was identified as a member of the genus Thermotoga on the basis of the presence of the typical outer sheath-like structure (toga) and 16S rRNA signature sequences and its ability to grow on carbohydrates (glucose, arabinose, fructose, lactose, maltose, and xylose). Major differences in its 16S rRNA gene sequence, its lower optimum temperature for growth (66 degrees C), its sodium chloride range for growth (0 to 2.8%), its lack of lactate as an end product from glucose fermentation, and its peritrichous flagella indicate that strain SEBR 6459T is not similar to the three previously described Thermotoga species. Furthermore, this organism does not belong to any of the other genera related to the order Thermotogales that have been described. On the basis of these findings, we propose that this strain should be described as a new species, Thermotoga elfii. The type strain of T. elfii is SEBR 6459 (= DSM 9442).

International audience; The effect of pH on the growth and physiology of Bacillus thermoamylovorans, a new moderately thermophilic and non-spore-forming bacterium isolated from palm wine, was studied. Growth occurred from pH 5.4 to 8.5, with optimum growth at 7.0. During the exponential growth phase at optimum pH, glucose was consumed at the maximum rate (qs), 17.87 mmol g(sup-1) h(sup-1), and was mainly fermented into acetate, ethanol, and formate (76.5% of metabolites produced). In acidic or alkaline conditions, glucose specific consumption rates were considerably reduced (qs = 8.06 mmol g(sup-1) h(sup-1) at pH 5.6 and 2.85 mmol g(sup-1) h(sup-1) at pH 8.4), and a switch in glucose metabolism toward lactate production (62.6% of metabolites produced at pH 5.6 and 41.2% of those produced at pH 8.4) was observed. Moreover, optimum cellular yield (Y(infx/ATP)), 14.8 g mol(sup-1), and optimum energy yield (Y(infATP/s)), 2.65 mol mol(sup-1), were observed at neutrality. The results of this study were compared with published data about lactic acid bacteria; this comparison allowed us to complement our previous taxonomic study of B. thermoamylovorans and to identify additional phenotypic differences between B. thermoamylovorans and lactobacilli.

The occurrence of thermophilic, halophilic anaerobic bacteria in the sediment of a Tunisian salted lake was tested in samples collected at 20-cm intervals down to a depth of 1.20 m. A long rod, present only in the 40- to 60-cm layer, was isolated at 60 degrees C in a medium containing 100 g of NaCl per liter and designated strain H168. This strain produced acetate, ethanol, H2, and CO2 from glucose metabolism. Fructose, xylose, ribose, cellobiose, and starch were also oxidized. The optimum temperature for growth was 60 degrees C. No growth was obtained at 42 or 70 degrees C. Strain H168 grew optimally in NaCl concentrations ranging from 50 to 100 g per liter, with the upper and lower limits of growth around 200 and 40 g per liter, respectively. The G+C ratio of the DNA was 39.6 mol%. Although halophilic, moderately thermophilic bacteria have been characterized among anaerobes, particularly within methanogens, strain H168 is the first true thermophilic (growing above 60 degrees C) halophilic anaerobic bacterium described so far. The phylogeny, physiology, morphology, lipid content, and high G+C content of strain H168 are sufficiently different from those of genera belonging to the family Haloanaerobiaceae to justify the definition of a new genus.

Summary A new thermophilic species of the genus Clostridium is described, which was isolated from palm wine in Senegal. The cells were anaerobic sporeforming rods, 0.7–1.0 μm wide and 2.0 to 8.0 μm long. The optimum growth temperature was about 55 °C. Sugars were mainly fermented to butyric acid. H 2 , CO 2 , small amounts of acetate, lactate and ethanol were produced. The G+C content of DNA was 35.7±0.3 mol%. The name Clostridium thermopalmarium sp. nov. is proposed for this species; the type strain is DSM 5974.

Thermophilic degradation of sugar beet pulp was studies in batch cultures at 55°C by different associations of bacteria, including CLOSTRIDIUM THERMOCELLUM, METHANOBACTERIUM sp. and METHANOSARCINA MP. C. THERMOCELLUM produced acetate, succinate, methanol, ethanol, H2 and CO2. The coculture of C. THERMOCELLUM and METHANOBACTERIUM sp. produced trace amounts of ethanol and succinate ; acetate concentration was about three times higher than in the C. THERMOCELLUM monoculture. The association of this coculture with METHANOSARCINA MP produced 5.5 mmol CH4/g dry weight sugar beet pulp. (Résumé d'auteur)

A thermophilic Methanosarcina strain was isolated from a digester fed with water hyacinths and inoculated with ground termites from the Congo. Optimal growth temperature was 55 degrees C. Methane production was at its optimum between pH 6.5 and 7.0. The bacterium grew on acetate, methanol and methylamines in the absence of growth factors, but could not use H2-CO2 or formate. H2-CO2 inhibited acetate utilisation. Yeast extract and vitamins stimulated growth.

A strictly anaerobic, thermophilic, fatty acids-degrading, sporulating sulfate-reducing bacterium was isolated from geothermal ground water. The organism stained Gram-negative and formed gas vacuoles during sporulation. Lactate, ethanol, fructose and saturated fatty acids up to C18 served as electron donors and carbon sources with sulfate as external electron acceptor. Benzoate was not used. Stoichiometric measurements revealed a complete oxidation of part of butyrate although growth with acetate as only electron donor was not observed. The rest of butyrate was oxidized to acetate. The strain grew chemolithoautotrophically with hydrogen plus sulfate as energy source and carbon dioxide as carbon source without requirement of additional organic carbon like acetate. The strain contained a c-type cytochrome and presumably a sulfite reductase P582. Optimum temperature, pH and NaCl concentration for growth were 54 degrees C, pH 7.3-7.5 and 25 to 35 g NaCl/l. The G + C content of DNA was 50.4 mol %. Strain BSD is proposed as a new species of the spore-forming sulfate-reducing genus Desulfotomaculum, D. geothermicum.

Ce travail décrit l'étude de la biométhanisation d'un résidu solide d'industrie agroalimentaire, les pulpes de betterave sucrière, par une culture mixte de bactéries thermophiles. Ces pulpes sont composées en grande partie de cellulose, de protéines et de pectines. La première phase a permis l'isolement de bactéries acidifiantes et méthanogènes thermophiles dont certaines ont été associées dans des cultures mixtes définies. Une bactérie protéolytique, #Thermobacteroides proteolyticus$, et trois bactéries méthanogènes appartenant aux genres #Methanobacterium$, #Methanogenium$ et #Methanosarcina$ ont été identifiées. Dans une deuxième étape, une expérimentation en cultures mixtes définies a été réalisée pour méthaniser la gélatine en associant #Thermobacteroides proteolyticus$ à #Methanobacterium$ sp. et #Methanosarcina$ MP. Ces mêmes bactéries méthanogènes ont été couplées à #Clostridium thermocellum$ pour la fermentation méthanique de la cellulose et des pectines. L'ensemble de ces bactéries ont été associées pour la méthanisation des pulpes de betterave sucrière avec un rendement de 5,5 mmole de méthane par g de matière sèche. Ces résultats montrent qu'il existe un transfert interspécifique d'hydrogène entre les bactéries fermentaires et les bactéries méthanogènes sur gélatine et cellulose ; ce processus entraîne une modification du métabolisme de la bactérie hydrolytique impliquée. Au contraire, l'utilisation des pectines ne conduit à aucun transfert d'H2 obligé. Cette étude démontre, d'autre part, que l'utilisation de l'acétate par #Methanosarcina$ MP dans la culture mixte n'est réalisable qu'en l'absence d'hydrogène. (Résumé d'auteur)

The fermentation of cellulose at 55°C by different associations of the 3 bacteria CLOSTRIDIUM THERMOCELLUM, METHANOBACTERIUM sp. and METHANOSARCINA MP, was studied. C. THERMOCELLUM alone produced acetate, lactate, ethanol, H2 and CO2. The co-culture C. THERMOCELLUM-METHANOBACTERIUM sp. produced more acetate and less ethanol than the monoculture of CLOSTRIDIUM. METHANOSARCINA MP used acetate only in the triculture including METHANOBACTERIUM sp. When methanol was added (5 mM) to the triculture, METHANOSARCINA MP had a shorter lag phase on acetate and degraded much more acetate. Maximum methane production was 8.5 mmol CH4/g cellulose degraded. (Résumé d'auteur)

La méthanisation d'#Euphorbia tirucalli$ a été réalisée en conditions mésophile et modérément thermophile. Trois différents pré-traitements de cette plante ont été testés : simple fragmentation, broyat, séchage puis broyage. Les digesteurs ont été alimentés avec 5 % de matière sèche en batch. Lors d'une expérimentation de 35 jours, le meilleur rendement (0,78 1/1/j) a été obtenu avec les fragments de plante en condition thermophile modérée. (Résumé d'auteur)