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Michel Drancourt - One of the best experts on this subject based on the ideXlab platform.
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Methanobrevibacter smithii tonsillar phlegmon a case report
new microbes and new infections, 2021Co-Authors: K Djemai, Michel Drancourt, Frederique Gouriet, J Michel, T Radulesco, Ghiles GrineAbstract:Abstract Untreated tonsillar phlegmon is a life-threatening condition commonly caused by Streptococcus pyogenes and Fusobacterium necrophorum among other pathogens. Here using specific laboratory tools, we detected Methanobrevibacter smithii in addition to S. pyogenes. This unprecedented observation questions the role of methanogens in phlegmon, and the optimal treatment of these mixed infection.
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Detection of Methanobrevobacter smithii and Methanobrevibacter oralis in Lower Respiratory Tract Microbiota
Microorganisms, 2020Co-Authors: Yasmine Hassani, Michel Drancourt, F. Bregeon, Gérard Aboudharam, Ghiles GrineAbstract:Methanogens, the sole microbes producing methane, are archaea commonly found in human anaerobic microbiota. Methanogens are emerging as opportunistic pathogens associated with dysbiosis and are also detected and cultured in anaerobic abscesses. Their presence in the respiratory tract is yet unknown. As a preliminary answer, prospective investigation of 908 respiratory tract samples using polyphasic approach combining PCR-sequencing, real-time PCR, fluorescent in situ hybridization (FISH), and methanogens culture was carried out. Methanobrevibacter smithii and Methanobrevibacter oralis DNA sequences, were detected in 21/527 (3.9%) sputum samples, 2/188 (1.06%) bronchoalveolar lavages, and none of 193 tracheo-bronchial aspirations. Further, fluorescence in situ hybridization detected methanogens in three sputum investigated specimens with stick morphology suggesting M. oralis and in another one bronchoalveolar lavage sample investigated, diplococal morphology suggesting M. smithii. These observations extend the known territory of methanogens to the respiratory tract and lay the foundations for further interpretation of their detection as pathogens in any future cases of isolation from bronchoalveolar lavages and the lungs.
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Culture of Methanogenic Archaea from Human Colostrum and Milk
Scientific reports, 2019Co-Authors: Amadou Hamidou Togo, Saber Khelaifia, C. Robert, Marion S. Bonnet, Ghiles Grine, Véronique Brevaut, Aurelia Caputo, Emeline Baptiste, Anthony Levasseur, Michel DrancourtAbstract:Archaeal sequences have been detected in human colostrum and milk, but no studies have determined whether living archaea are present in either of these fluids. Methanogenic archaea are neglected since they are not detected by usual molecular and culture methods. By using improved DNA detection protocols and microbial culture techniques associated with antioxidants previously developed in our center, we investigated the presence of methanogenic archaea using culture and specific Methanobrevibacter smithii and Methanobrevibacter oralis real-time PCR in human colostrum and milk. M. smithii was isolated from 3 colostrum and 5 milk (day 10) samples. M. oralis was isolated from 1 milk sample. For 2 strains, the genome was sequenced, and the rhizome was similar to that of strains previously isolated from the human mouth and gut. M. smithii was detected in the colostrum or milk of 5/13 (38%) and 37/127 (29%) mothers by culture and qPCR, respectively. The different distribution of maternal body mass index according to the detection of M. smithii suggested an association with maternal metabolic phenotype. M. oralis was not detected by molecular methods. Our results suggest that breastfeeding may contribute to the vertical transmission of these microorganisms and may be essential to seed the infant’s microbiota with these neglected critical commensals from the first hour of life.
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Nine Cases of Methanogenic Archaea in Refractory Sinusitis, an Emerging Clinical Entity
Frontiers Media S.A., 2019Co-Authors: Elisabeth Sogodogo, Vanessa Demonfort Nkamga, Mustapha Fellag, Ahmed Loukil, Justin Michel, Patrick Dessi, Pierre-edouard Fournier, Michel DrancourtAbstract:The authors report the cases of 9 patients eventually diagnosed with methanogenic archaea refractory or recalcitrant chronic rhinosinusitis, a condition known to involve various anaerobic bacteria but in which the role of methanogenic archaea is unknown. The authors retrospectively searched these microorganisms by PCR in surgically-collected sinusal pus specimens from patients diagnosed with refractory sinusitis, defined by the persistance of sinus inflammation and related-symptoms for more than 12 weeks despite appropriate treatment. Of the 116 tested sinus surgical specimens, 12 (10.3%) from 9 patients (six females, three males; aged 20–71 years) were PCR-positive. These specimens were further investigated by fluorescence in-situ hybridization, PCR amplicon-sequencing and culture. Methanobrevibacter smithii was documented in four patients and Methanobrevibacter oralis in another four, one of whom was also culture-positive. They were associated with a mixed flora including Gram-positive and Gram-negative bacteria. In the latter patient, “Methanobrevibacter massiliense” was the sole microorganism detected. These results highlight methanogenic archaea as being part of a mixed anaerobic flora involved in refractory sinusitis, and suggest that the treatment of this condition should include an antibiotic active against methanogens, notably a nitroimidazole derivative
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Archaea: Essential inhabitants of the human digestive microbiota
Human Microbiome Journal, 2017Co-Authors: Vanessa Demonfort Nkamga, Bernard Henrissat, Michel DrancourtAbstract:Prokaryotes forming the domain of Archaea, named after their first discovery in extreme environments, are acknowledged but still neglected members of the human digestive tract microbiota. In this microbiota, cultured archaea comprise anaerobic methanogens: Methanobrevibacter smithii, Methanobrevibacter oralis, Methanobrevibacter massiliense, Methanosphaera stadtmanae, Methanobrevibacter arboriphilus, Methanobrevibacter millerae and Methanomassiliicoccus luminyensis; along with the non-methanogen halophilic Archaea Halopherax massiliense. Metagenomic analyses detected DNA sequences indicative of the presence of additional methanogenic and non-methanogenic halophilic Archaea in the human intestinal tract and oral cavity. Methanogens specifically metabolize hydrogen produced by anaerobic fermentation of carbohydrates into methane; further transforming heavy metals and metalloids into methylated derivatives, such as trimethylbismuth which is toxic for both human and bacterial cells. However, the role of Archaea as pathogens remains to be established. Future researches will aim to increase the repertoire of the human digestive tract Archaea and to understand their possible association with intestinal and extra-intestinal infections and diseases including weight regulation abnormalities.
Akihiko Yamagishi - One of the best experts on this subject based on the ideXlab platform.
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Endosymbiotic Methanobrevibacter species Living in Symbiotic Protists of the Termite Reticulitermes speratus Detected by Fluorescent In Situ Hybridization
Microbes and Environments, 2004Co-Authors: Kurt Hara, Naoya Shinzato, Tairo Oshima, Akihiko YamagishiAbstract:Two species of cellulolytic protist, Dinenympha parva and Microjoenia, living in the guts of the lower termite Reticulitermes speratus are known to harbor endosymbiotic methanogens detectable with an epifluorescent microscope. DNA isolated from the guts of worker termites in a colony of R. speratus was amplified using archaea-specific primers and cloned, and partial 16S rRNA gene sequences were obtained. Archaeal PCR clones obtained from the guts of xylophagous insects in this and previous works formed four subgroups within the Methanobrevibacter branch of a phylogenetic tree; the sequences of the clones obtained in this report belonged to subgroups, designated XSAT1A and XSAT1D. Using a probe specific to each of the subgroups, 50 and 10 endosymbiotic Methanobrevibacter cells per protist respectively were detected with a probe specific to the subgroup XSAT1A by fluorescent in situ hybridization analysis in D. parva and Microjoenia. There was no observed hybridization to the endosymbiont with other subtype-specific probes including XSAT1D. Based on these results, endosymbionts in D. parva and Microjoenia sp. are proposed to belong to the Methanobrevibacter subgroup XSAT1A.
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phylogenetic diversity of symbiotic methanogens living in the hindgut of the lower termite reticulitermes speratus analyzed by pcr and in situ hybridization
Applied and Environmental Microbiology, 1999Co-Authors: Naoya Shinzato, Tadao Matsumoto, Ikuo Yamaoka, Tairo Oshima, Akihiko YamagishiAbstract:A phylogenetic analysis of the sequences of 60 clones of archaeal small-subunit rRNA genes amplified from the termite Reticulitermes speratus revealed that most of them (56 clones) clustered in the genus Methanobrevibacter. Three clones were classified in the order Thermoplasmales. The Methanobrevibacter-related symbionts were detected by in situ hybridization analysis.
Andredenis G Wright - One of the best experts on this subject based on the ideXlab platform.
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response of the rumen microbiota of sika deer cervus nippon fed different concentrations of tannin rich plants
PLOS ONE, 2015Co-Authors: Zhipeng Li, Andredenis G Wright, Fuhe Yang, Zhigang Zhang, Guangyu LiAbstract:High throughput sequencing was used to examine the rumen microbiota of sika deer fed high (OLH) and low concentration (OLL) of tannin rich oak leaves. The results showed that Prevotella spp. were the most dominant bacteria. The most predominant methanogens were the members of the order Methanoplasmatales. The dominant rumen protozoa were Entodinium longinucleatum, Eudiplodinium maggii, and Epidinium caudatum, and the fungal communities were mostly represented by Piromyces spp. Moreover, the relative abundance of Pseudobutyrivibrio spp. (P=0.026), unidentified bacteria (P=0.028), and Prevotella spp. (P=0.022) was lower in the OLH group than in the OLL group. The concentration of propionate in the OLH group was greater than in the OLL group (P=0.006). Patterns of relationships showed that methanogens belonging to the order Methanoplasmatales were negatively correlated with Treponema spp., Ent. Longinucleatum, and acetate. Methanosphaera stadtmanae was positively correlated to propionate, while Methanobrevibacter ruminantium was negatively associated with Methanobrevibacter thaueri and Methanobrevibacter millerae. Tannins altered the rumen microbes and fermentation patterns. However, the response of the entire rumen microbiota and the relationship between rumen microorganisms and the fermentation parameters were not fully understood.
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Molecular analysis of methanogenic archaea in the forestomach of the alpaca (Vicugna pacos)
BMC Microbiology, 2012Co-Authors: Benoit St-pierre, Andredenis G WrightAbstract:Background Methanogens that populate the gastrointestinal tract of livestock ruminants contribute significantly to methane emissions from the agriculture industry. There is a great need to analyze archaeal microbiomes from a broad range of host species in order to establish causal relationships between the structure of methanogen communities and their potential for methane emission. In this report, we present an investigation of methanogenic archaeal populations in the foregut of alpacas. Results We constructed individual 16S rRNA gene clone libraries from five sampled animals and recovered a total of 947 sequences which were assigned to 51 species-level OTUs. Individuals were found to each have between 21 and 27 OTUs, of which two to six OTUs were unique. As reported in other host species, Methanobrevibacter was the dominant genus in the alpaca, representing 88.3% of clones. However, the alpaca archaeal microbiome was different from other reported host species, as clones showing species-level identity to Methanobrevibacter millerae were the most abundant. Conclusion From our analysis, we propose a model to describe the population structure of Methanobrevibacter -related methanogens in the alpaca and in previously reported host species, which may contribute in unraveling the complexity of symbiotic archaeal communities in herbivores.
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Rumen-like methanogens identified from the crop of the folivorous South American bird, the hoatzin (Opisthocomus hoazin)
The ISME Journal, 2009Co-Authors: Andredenis G Wright, Korinne S Northwood, Nestor E ObispoAbstract:The hoatzin is the only known avian species with foregut fermentation. It is a primarily folivorous feeder and has a distended crop and lower/distal esophagus, which has evolved for the microbial fermentation of ingested feed. Crop samples collected from 10 individual animals from the Apure River area, Apure State, Venezuela were examined for the presence and density of methanogens using 16S rRNA gene clone libraries and real-time PCR prepared from pooled and individual PCR products. A total of 197 clones were examined, revealing 24 different methanogen 16S rRNA sequences, or phylotypes. Of the 24 unique phylotypes, 16 (171 of 197 clones) formed five unique clades within the genus Methanobrevibacter with the largest group of clones (118 clones) 98.7% similar to Methanobrevibacter ruminantium . The remaining eight phylotypes (26 clones) formed four unique clades that had only 94.0–96.7% identity to Methanosphaera stadtmanae . Based upon 98% sequence identity, we identified 17 of the 24 methanogen phylotypes from the hoatzin as possible new species and strains, with three phylotypes representing possible new genera (
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"Methanobrevibacter" phylotypes are the dominant methanogens in sheep from Venezuela
Microbial ecology, 2007Co-Authors: Andredenis G Wright, Nestor E ObispoAbstract:Rumen methanogens in sheep from Venezuela were examined using 16S rRNA gene libraries and denaturing gradient gel electrophoresis (DGGE) profiles prepared from pooled and individual PCR products from the rumen contents from 10 animals. A total of 104 clones were examined, revealing 14 different 16S rRNA gene sequences or phylotypes. Of the 14 phylotypes, 13 (99 of 104 clones) belonged to the genus Methanobrevibacter, indicating that the genus Methanobrevibacter is the most dominant component of methanogen populations in sheep in Venezuela. The largest group of clones (41 clones) was 97.9-98.5% similar to Methanobrevibacter gottschalkii. Two sequences were identified as possible new species, one belonging to the genus Methanobrevibacter and the other belonging to the genus Methanobacterium. DGGE analysis of the rumen contents from individual animals also revealed 14 different bands with a range of 4-9 bands per animal.
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Methanobrevibacter millerae sp nov and Methanobrevibacter olleyae sp nov methanogens from the ovine and bovine rumen that can utilize formate for growth
International Journal of Systematic and Evolutionary Microbiology, 2007Co-Authors: J P Bowman, Carolyn Pimm, Sam Popovski, Andredenis G WrightAbstract:Four formate-utilizing methanogens were isolated from ovine (strain KM1H5-1P T ) and bovine (strains AK-87, OCP and ZA-10 T ) rumen contents. Based on 16S rRNA gene sequence analysis, the methanogen strains were found to belong to the order Methanobacteriales in the genus Methanobrevibacter.Strains ZA-10 T and KM1H5-1P T gained energy for growth by the reduction of CO2 to CH4 using H2 or formate exclusively as electron donors. Increasing formate concentrations to 220 mM in batch cultures increased the growth of strain KM1H5-1P T but did not affect the growth of strain ZA-10 T . Substrate specificity and resistance to cell-wall lysis supported the affiliation of the strains to the genus Methanobrevibacter. Strains ZA-10 T and KM1H5-1P T showed 16S rRNA gene sequence similarity of 98.0 and 98.6% to their closest recognized relatives, Methanobrevibacter thaueri CW T and Methanobrevibacter ruminantium M1 T , respectively. DNA‐DNA hybridization experiments indicated that the strains were not affiliated at the species level to their closest recognized relatives, with DNA reassociation values of only 28% between strains ZA-10 T and Methanobrevibacter thaueri CW T and <25% between strains KM1H5-1P T and Methanobrevibacter ruminantium M1 T . Based on the data presented, the new strains are considered to represent two novel species of the genus Methanobrevibacter, for which the names Methanobrevibacter millerae sp. nov. (type strain ZA-10 T =DSM 16643 T =OCM 820 T ) and Methanobrevibacter olleyae sp. nov. (type strain KM1H5-1P T =DSM 16632 T =OCM 841 T ) are proposed.
Naoya Shinzato - One of the best experts on this subject based on the ideXlab platform.
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Application of pseudomurein endoisopeptidase to fluorescence in situ hybridization of methanogens within the family Methanobacteriaceae.
Applied and Environmental Microbiology, 2006Co-Authors: Kohei Nakamura, Xian-ying Meng, Miho Enoki, Naoya Shinzato, Yuji Sekiguchi, Takeshi Terada, Yoichi KamagataAbstract:In situ detection of methanogens within the family Methanobacteriaceae is sometimes known to be unsuccessful due to the difficulty in permeability of oligonucleotide probes. Pseudomurein endoisopeptidase (Pei), a lytic enzyme that specifically acts on their cell walls, was applied prior to 16S rRNA-targeting fluorescence in situ hybridization (FISH). For this purpose, pure cultured methanogens within this family, Methanobacterium bryantii, Methanobrevibacter ruminantium, Methanosphaera stadtmanae, and Methanothermobacter thermautotrophicus together with a Methanothermobacter thermautotrophicus-containing syntrophic acetate-oxidizing coculture, endosymbiotic Methanobrevibacter methanogens within an anaerobic ciliate, and an upflow anaerobic sludge blanket (UASB) granule were examined. Even without the Pei treatment, Methanobacterium bryantii and Methanothermobacter thermautotrophicus cells are relatively well hybridized with oligonucleotide probes. However, almost none of the cells of Methanobrevibacter ruminantium, Methanosphaera stadtmanae, cocultured Methanothermobacter thermautotrophicus, and the endosymbiotic methanogens and the cells within UASB granule were hybridized. Pei treatment was able to increase the probe hybridization ratio in every specimen, particularly in the specimen that had shown little hybridization. Interestingly, the hybridizing signal intensity of Methanothermobacter thermautotrophicus cells in coculture with an acetate-oxidizing H2-producing syntroph was significantly improved by Pei pretreatment, whereas the probe was well hybridized with the cells of pure culture of the same strain. We found that the difference is attributed to the differences in cell wall thicknesses between the two culture conditions. These results indicate that Pei treatment is effective for FISH analysis of methanogens that show impermeability to the probe.
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Endosymbiotic Methanobrevibacter species Living in Symbiotic Protists of the Termite Reticulitermes speratus Detected by Fluorescent In Situ Hybridization
Microbes and Environments, 2004Co-Authors: Kurt Hara, Naoya Shinzato, Tairo Oshima, Akihiko YamagishiAbstract:Two species of cellulolytic protist, Dinenympha parva and Microjoenia, living in the guts of the lower termite Reticulitermes speratus are known to harbor endosymbiotic methanogens detectable with an epifluorescent microscope. DNA isolated from the guts of worker termites in a colony of R. speratus was amplified using archaea-specific primers and cloned, and partial 16S rRNA gene sequences were obtained. Archaeal PCR clones obtained from the guts of xylophagous insects in this and previous works formed four subgroups within the Methanobrevibacter branch of a phylogenetic tree; the sequences of the clones obtained in this report belonged to subgroups, designated XSAT1A and XSAT1D. Using a probe specific to each of the subgroups, 50 and 10 endosymbiotic Methanobrevibacter cells per protist respectively were detected with a probe specific to the subgroup XSAT1A by fluorescent in situ hybridization analysis in D. parva and Microjoenia. There was no observed hybridization to the endosymbiont with other subtype-specific probes including XSAT1D. Based on these results, endosymbionts in D. parva and Microjoenia sp. are proposed to belong to the Methanobrevibacter subgroup XSAT1A.
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phylogenetic diversity of symbiotic methanogens living in the hindgut of the lower termite reticulitermes speratus analyzed by pcr and in situ hybridization
Applied and Environmental Microbiology, 1999Co-Authors: Naoya Shinzato, Tadao Matsumoto, Ikuo Yamaoka, Tairo Oshima, Akihiko YamagishiAbstract:A phylogenetic analysis of the sequences of 60 clones of archaeal small-subunit rRNA genes amplified from the termite Reticulitermes speratus revealed that most of them (56 clones) clustered in the genus Methanobrevibacter. Three clones were classified in the order Thermoplasmales. The Methanobrevibacter-related symbionts were detected by in situ hybridization analysis.
Saber Khelaifia - One of the best experts on this subject based on the ideXlab platform.
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Culture of Methanogenic Archaea from Human Colostrum and Milk
Scientific reports, 2019Co-Authors: Amadou Hamidou Togo, Saber Khelaifia, C. Robert, Marion S. Bonnet, Ghiles Grine, Véronique Brevaut, Aurelia Caputo, Emeline Baptiste, Anthony Levasseur, Michel DrancourtAbstract:Archaeal sequences have been detected in human colostrum and milk, but no studies have determined whether living archaea are present in either of these fluids. Methanogenic archaea are neglected since they are not detected by usual molecular and culture methods. By using improved DNA detection protocols and microbial culture techniques associated with antioxidants previously developed in our center, we investigated the presence of methanogenic archaea using culture and specific Methanobrevibacter smithii and Methanobrevibacter oralis real-time PCR in human colostrum and milk. M. smithii was isolated from 3 colostrum and 5 milk (day 10) samples. M. oralis was isolated from 1 milk sample. For 2 strains, the genome was sequenced, and the rhizome was similar to that of strains previously isolated from the human mouth and gut. M. smithii was detected in the colostrum or milk of 5/13 (38%) and 37/127 (29%) mothers by culture and qPCR, respectively. The different distribution of maternal body mass index according to the detection of M. smithii suggested an association with maternal metabolic phenotype. M. oralis was not detected by molecular methods. Our results suggest that breastfeeding may contribute to the vertical transmission of these microorganisms and may be essential to seed the infant’s microbiota with these neglected critical commensals from the first hour of life.
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Aerobic culture of methanogenic archaea without an external source of hydrogen
European Journal of Clinical Microbiology and Infectious Diseases, 2016Co-Authors: Saber Khelaifia, Vanessa Demonfort Nkamga, Jean Christophe Lagier, Elodie GuilhotAbstract:Culturing methanogenic archaea is fastidious, ex-pensive, and requires an external source of hydrogen and car-bon dioxide. Until now, these microorganisms have only been cultivated under strictly anaerobic conditions. We previously developed a single versatile culture medium containing sugars and anti-oxydants for cultivating all human known methanogens. Performing aerobic cultures in the presence of Bacteroides thetaiotaomicron, which produces hydrogen, al-lows for cultivation of Methanobrevibacter smithii which itself produces methane. To obtain colonies, we cultivated M. smithii in an agar plate in the upper part of a double chamber flask with a liquid culture of B. thetaiotaomicron in the lower compart-ment. We subsequently cultured four other methanogenic spe-cies for the first time and successfully isolated 13 strains of M. smithii and nine strains of Methanobrevibacter oralis from 100 stools and 45 oral samples. This procedure allows aerobic iso-lation and antibiotic susceptibility testing. This changes the ability to routinely study methanogens, which have been neglected in clinical microbiology laboratories and may be use-ful for biogas production.
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Draft Genome Sequencing of Methanobrevibacter oralis Strain JMR01, Isolated from the Human Intestinal Microbiota
Genome announcements, 2014Co-Authors: Saber Khelaifia, Didier Raoult, Marc Garibal, Catherine Robert, Michel DrancourtAbstract:Methanobrevibacter oralis, an anaerobic methanogenic archaeon, has been previously isolated from the human oral cavity. Here, sequencing a stool isolate (strain JMR01) yielded a 2.065-Mb genome with a 27.78% G+C content containing a total of 2,042 open reading frames and 3 clusters of regularly interspaced short palindromic repeat (CRISPR) loci with associated Cas proteins.
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Draft Genome Sequence of a Human-Associated Isolate of Methanobrevibacter arboriphilicus, the Lowest-G+C-Content Archaeon
Genome Announcements, 2014Co-Authors: Saber Khelaifia, Marc Garibal, Catherine Robert, Michel DrancourtAbstract:We report the draft genome sequence of Methanobrevibacter arboriphilicus strain ANOR1, isolated from the human gut. Its 2.21-Mb genome exhibits a 25.46% GC content, the lowest value among archaea. The genome of M. arboriphilicus contains a total of 2,111 open reading frames and three clusters of regularly interspaced short palindromic repeat (CRISPR) loci with associated Cas proteins. Citation Khelaifia S, Garibal M, Robert C, Raoult D, Drancourt M. 2014. Draft genome sequence of a human-associated isolate of Methanobrevibacter arboriphilicus, the lowest-GC-content archaeon. Genome Announc. 2(1):e01181-13.
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Draft Genome Sequencing of Methanobrevibacter oralis Strain JMR01, Isolated from the Human Intestinal Microbiota
Genome Announcements, 2014Co-Authors: Rita Aghnatios, Michel Drancourt, Didier Raoult, Marc Garibal, Catherine Robert, Caroline Cayrou, Said Azza, Saber KhelaifiaAbstract:Methanobrevibacter oralis, an anaerobic methanogenic archaeon, has been previously isolated from the human oral cavity. Here, sequencing a stool isolate (strain JMR01) yielded a 2.065-Mb genome with a 27.78% GC content containing a total of 2,042 open reading frames and 3 clusters of regularly interspaced short palindromic repeat (CRISPR) loci with associated Cas proteins. Address correspondence to Michel Drancourt, michel.drancourt@univ-amu.fr. M ethanobrevibacter oralis was initially isolated from human subgingival plaque (1), and further studies associated this methanogen with subgingival diseases, including periodontitis (2). We recently demonstrated a significant correlation between the M. oralis charge and the severity of periodontitis (3). Furthermore , M. oralis DNA was detected in the human gut, and we previously reported the first isolation of M. oralis (strain JMR01) from the human gut (4). M. oralis strain JMR01 is a strict anaer-obic archaeon with optimal growth at 37°C (pH 7.5) and 1.5 g/liter NaCl. Methane is produced by reducing acetate or H 2-CO 2 as an electron donor. Rumen fluid and vitamin solutions are required for growth and a volatile fatty acid mixture stimulates growth. Phylogenetic classification based on the 16S rRNA gene sequence (GenBank accession number KC616346) confirmed the affiliation of the M. oralis strain JMR01 herein sequenced with the genus Methanobrevibacter and yielded a sequence similarity of 99% with the reference strain M. oralis DSM 7256 (GenBank accession number HE654003). The M. oralis strain JMR01 complete genome was sequenced by combining shotgun and 3-kb paired-end libraries using high-throughput 454 pyrosequencing (454 Life Sciences-Roche, Boulogne-Billancourt, France). Sequence reads were assembled using a Newbler assembler 2.8 (20120726_1306) (Roche), and 136 contigs were generated into 14 scaffolds. A preliminary open reading frame (ORF) prediction was conducted by automated annotation with Glimmer (http://www .cbcb.umd.edu/software/glimmer/) and RAST (5). The annotation was manually curated using BLAST and the nr database of NCBI. The CRISPR finder (http://crispr.u-psud.fr/Server/) was used to detect and identify CRISPR repeat and spacer sequences in the genome. The M. oralis genome consists of a 2,065,764-bp circular chromosome (with a GC content of 27.78%). A total of 2,042 ORFs were recovered, encoding proteins involved in functions previously assigned in other related Methanobrevibacter organisms (6). Likewise, the M. oralis genome contains three CRISPR loci and associated proteins (Cas), as previously found in Methanobre-vibacter arboriphilicus (7). We previously observed that M. oralis, an inhabitant of the oral cavity, survives in the acidity of the stomach (S. Khelaifia and M. Drancourt, unpublished data), giving it an opportunity to establish itself in the digestive tract. Accordingly, we isolated M. oralis from the human gut. The availability of the M. oralis genome will give the opportunity to develop genotyping methods to investigate the diversity of this frequently detected human archaeon. Moreover, the isolation of a new M. oralis strain from the human gut, as well as its genome sequencing, can increase knowledge of this neglected component of the human gut microbiota. Nucleotide sequence accession numbers. The Methanobre-vibacter oralis strain JMR01 genome sequence has been deposited in EMBL under the accession numbers CBWS010000001 through CBWS010000060. The whole-genome shotgun project has been deposited in GenBank under the accession number CBWS000000000. ACKNOWLEDGMENT