Mycobacterium abscessus virulence traits unraveled by transcriptomic profiling in amoeba and macrophages
Identifieur interne : 000B60 ( Main/Exploration ); précédent : 000B59; suivant : 000B61Mycobacterium abscessus virulence traits unraveled by transcriptomic profiling in amoeba and macrophages
Auteurs : Violaine Dubois [France] ; Alexandre Pawlik [France] ; Anouchka Bories [France] ; Vincent Le Moigne [France] ; Odile Sismeiro [France] ; Rachel Legendre [France] ; Hugo Varet [France] ; María Del Pilar Rodríguez-Ord Ez [France] ; Jean-Louis Gaillard [France] ; Jean-Yves Coppée [France] ; Roland Brosch [France] ; Jean-Louis Herrmann [France] ; Fabienne Girard-Misguich [France]Source :
- PLoS Pathogens [ 1553-7366 ] ; 2019.
Abstract
Free-living amoebae are thought to represent an environmental niche in which amoeba-resistant bacteria may evolve towards pathogenicity. To get more insights into factors playing a role for adaptation to intracellular life, we characterized the transcriptomic activities of the emerging pathogen
Url:
DOI: 10.1371/journal.ppat.1008069
PubMed: 31703112
PubMed Central: 6839843
Affiliations:
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type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff004"><addr-line>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Varet, Hugo" sort="Varet, Hugo" uniqKey="Varet H" first="Hugo" last="Varet">Hugo Varet</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country 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GHU Paris Saclay, Hôpital Ambroise Paré, Boulogne Billancourt, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>AP-HP. GHU Paris Saclay, Hôpital Ambroise Paré, Boulogne Billancourt</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Boulogne-Billancourt</settlement></placeName></affiliation></author><author><name sortKey="Coppee, Jean Yves" sort="Coppee, Jean Yves" uniqKey="Coppee J" first="Jean-Yves" last="Coppée">Jean-Yves Coppée</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Brosch, Roland" sort="Brosch, Roland" uniqKey="Brosch R" first="Roland" last="Brosch">Roland Brosch</name><affiliation wicri:level="3"><nlm:aff id="aff002"><addr-line>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Herrmann, Jean Louis" sort="Herrmann, Jean Louis" uniqKey="Herrmann J" first="Jean-Louis" last="Herrmann">Jean-Louis Herrmann</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff007"><addr-line>AP-HP. GHU Paris Saclay, Hôpital Raymond Poincaré, Garches, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>AP-HP. GHU Paris Saclay, Hôpital Raymond Poincaré, Garches</wicri:regionArea><wicri:noRegion>Garches</wicri:noRegion><wicri:noRegion>Garches</wicri:noRegion></affiliation></author><author><name sortKey="Girard Misguich, Fabienne" sort="Girard Misguich, Fabienne" uniqKey="Girard Misguich F" first="Fabienne" last="Girard-Misguich">Fabienne Girard-Misguich</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31703112</idno><idno type="pmc">6839843</idno><idno 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type="main"><italic>Mycobacterium abscessus</italic> virulence traits unraveled by transcriptomic profiling in amoeba and macrophages</title><author><name sortKey="Dubois, Violaine" sort="Dubois, Violaine" uniqKey="Dubois V" first="Violaine" last="Dubois">Violaine Dubois</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation></author><author><name sortKey="Pawlik, Alexandre" sort="Pawlik, Alexandre" uniqKey="Pawlik A" first="Alexandre" last="Pawlik">Alexandre Pawlik</name><affiliation wicri:level="3"><nlm:aff id="aff002"><addr-line>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Bories, Anouchka" sort="Bories, Anouchka" uniqKey="Bories A" first="Anouchka" last="Bories">Anouchka Bories</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation></author><author><name sortKey="Le Moigne, Vincent" sort="Le Moigne, Vincent" uniqKey="Le Moigne V" first="Vincent" last="Le Moigne">Vincent Le Moigne</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation></author><author><name sortKey="Sismeiro, Odile" sort="Sismeiro, Odile" uniqKey="Sismeiro O" first="Odile" last="Sismeiro">Odile Sismeiro</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Legendre, Rachel" sort="Legendre, Rachel" uniqKey="Legendre R" first="Rachel" last="Legendre">Rachel Legendre</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff004"><addr-line>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Varet, Hugo" sort="Varet, Hugo" uniqKey="Varet H" first="Hugo" last="Varet">Hugo Varet</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff004"><addr-line>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Transcriptome and Epigenome Platform—Biomics Pole—CITECH, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Rodriguez Ord Ez, Maria Del Pilar" sort="Rodriguez Ord Ez, Maria Del Pilar" uniqKey="Rodriguez Ord Ez M" first="María Del Pilar" last="Rodríguez-Ord Ez">María Del Pilar Rodríguez-Ord Ez</name><affiliation wicri:level="3"><nlm:aff id="aff005"><addr-line>Laboratoire d'Écologie, Systématique et Évolution, Université Paris-Saclay, Orsay, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire d'Écologie, Systématique et Évolution, Université Paris-Saclay, Orsay</wicri:regionArea><placeName><settlement type="city">Orsay</settlement></placeName></affiliation></author><author><name sortKey="Gaillard, Jean Louis" sort="Gaillard, Jean Louis" uniqKey="Gaillard J" first="Jean-Louis" last="Gaillard">Jean-Louis Gaillard</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation><affiliation wicri:level="3"><nlm:aff id="aff006"><addr-line>AP-HP. GHU Paris Saclay, Hôpital Ambroise Paré, Boulogne Billancourt, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>AP-HP. GHU Paris Saclay, Hôpital Ambroise Paré, Boulogne Billancourt</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Boulogne-Billancourt</settlement></placeName></affiliation></author><author><name sortKey="Coppee, Jean Yves" sort="Coppee, Jean Yves" uniqKey="Coppee J" first="Jean-Yves" last="Coppée">Jean-Yves Coppée</name><affiliation wicri:level="3"><nlm:aff id="aff003"><addr-line>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur—Bioinformatics and Biostatistics Hub—C3BI, USR 3756 IP CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Brosch, Roland" sort="Brosch, Roland" uniqKey="Brosch R" first="Roland" last="Brosch">Roland Brosch</name><affiliation wicri:level="3"><nlm:aff id="aff002"><addr-line>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Institut Pasteur, Unité de Pathogénomique Mycobactérienne intégrée, UMR3525 CNRS, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Herrmann, Jean Louis" sort="Herrmann, Jean Louis" uniqKey="Herrmann J" first="Jean-Louis" last="Herrmann">Jean-Louis Herrmann</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff007"><addr-line>AP-HP. GHU Paris Saclay, Hôpital Raymond Poincaré, Garches, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>AP-HP. GHU Paris Saclay, Hôpital Raymond Poincaré, Garches</wicri:regionArea><wicri:noRegion>Garches</wicri:noRegion><wicri:noRegion>Garches</wicri:noRegion></affiliation></author><author><name sortKey="Girard Misguich, Fabienne" sort="Girard Misguich, Fabienne" uniqKey="Girard Misguich F" first="Fabienne" last="Girard-Misguich">Fabienne Girard-Misguich</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux, France</addr-line></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Université Paris-Saclay, UVSQ, Inserm, Infection et inflammation, Montigny-Le-Bretonneux</wicri:regionArea><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion><wicri:noRegion>Montigny-Le-Bretonneux</wicri:noRegion></affiliation></author></analytic><series><title level="j">PLoS Pathogens</title><idno type="ISSN">1553-7366</idno><idno type="eISSN">1553-7374</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Free-living amoebae are thought to represent an environmental niche in which amoeba-resistant bacteria may evolve towards pathogenicity. To get more insights into factors playing a role for adaptation to intracellular life, we characterized the transcriptomic activities of the emerging pathogen <italic>Mycobacterium abscessus</italic> in amoeba and murine macrophages (Mϕ) and compared them with the intra-amoebal transcriptome of the closely related, but less pathogenic <italic>Mycobacterium chelonae</italic>. Data on up-regulated genes in amoeba point to proteins that allow <italic>M</italic>. <italic>abscessus</italic> to resist environmental stress and induce defense mechanisms, as well as showing a switch from carbohydrate carbon sources to fatty acid metabolism. For eleven of the most upregulated genes in amoeba and/or Mϕ, we generated individual gene knock-out <italic>M</italic>. <italic>abscessus</italic> mutant strains, from which ten were found to be attenuated in amoeba and/or Mϕ in subsequence virulence analyses. Moreover, transfer of two of these genes into the genome of <italic>M</italic>. <italic>chelonae</italic> increased the intra-Mϕ survival of the recombinant strain. One knock-out mutant that had the gene encoding Eis N-acetyl transferase protein (<italic>MAB_4532c</italic>) deleted, was particularly strongly attenuated in Mϕ. Taken together, <italic>M</italic>. <italic>abscessus</italic> intra-amoeba and intra-Mϕ transcriptomes revealed the capacity of <italic>M</italic>. <italic>abscessus</italic> to adapt to an intracellular lifestyle, with amoeba largely contributing to the enhancement of <italic>M</italic>. <italic>abscessus</italic> intra-Mϕ survival.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Lavania, M" uniqKey="Lavania M">M Lavania</name></author><author><name sortKey="Katoch, K" uniqKey="Katoch K">K Katoch</name></author><author><name sortKey="Katoch, Vm" uniqKey="Katoch V">VM Katoch</name></author><author><name sortKey="Gupta, Ak" uniqKey="Gupta A">AK Gupta</name></author><author><name sortKey="Chauhan, Ds" uniqKey="Chauhan D">DS Chauhan</name></author><author><name sortKey="Sharma, R" uniqKey="Sharma R">R Sharma</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Angenent, Lt" uniqKey="Angenent L">LT 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uniqKey="Vallenet D">D Vallenet</name></author><author><name sortKey="Engelen, S" uniqKey="Engelen S">S Engelen</name></author><author><name sortKey="Mornico, D" uniqKey="Mornico D">D Mornico</name></author><author><name sortKey="Cruveiller, S" uniqKey="Cruveiller S">S Cruveiller</name></author><author><name sortKey="Fleury, L" uniqKey="Fleury L">L Fleury</name></author><author><name sortKey="Lajus, A" uniqKey="Lajus A">A Lajus</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alexa, A" uniqKey="Alexa A">A Alexa</name></author><author><name sortKey="Rahnenfuhrer, J" uniqKey="Rahnenfuhrer J">J Rahnenfuhrer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Simeone, R" uniqKey="Simeone R">R Simeone</name></author><author><name sortKey="Sayes, F" uniqKey="Sayes F">F Sayes</name></author><author><name sortKey="Song, O" uniqKey="Song O">O Song</name></author><author><name sortKey="Groschel, Mi" uniqKey="Groschel M">MI 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sortKey="Gaillard, Jean Louis" sort="Gaillard, Jean Louis" uniqKey="Gaillard J" first="Jean-Louis" last="Gaillard">Jean-Louis Gaillard</name><name sortKey="Gaillard, Jean Louis" sort="Gaillard, Jean Louis" uniqKey="Gaillard J" first="Jean-Louis" last="Gaillard">Jean-Louis Gaillard</name><name sortKey="Girard Misguich, Fabienne" sort="Girard Misguich, Fabienne" uniqKey="Girard Misguich F" first="Fabienne" last="Girard-Misguich">Fabienne Girard-Misguich</name><name sortKey="Herrmann, Jean Louis" sort="Herrmann, Jean Louis" uniqKey="Herrmann J" first="Jean-Louis" last="Herrmann">Jean-Louis Herrmann</name><name sortKey="Herrmann, Jean Louis" sort="Herrmann, Jean Louis" uniqKey="Herrmann J" first="Jean-Louis" last="Herrmann">Jean-Louis Herrmann</name><name sortKey="Le Moigne, Vincent" sort="Le Moigne, Vincent" uniqKey="Le Moigne V" first="Vincent" last="Le Moigne">Vincent Le Moigne</name><name sortKey="Legendre, Rachel" sort="Legendre, Rachel" uniqKey="Legendre R" first="Rachel" last="Legendre">Rachel Legendre</name><name sortKey="Legendre, Rachel" sort="Legendre, Rachel" uniqKey="Legendre R" first="Rachel" last="Legendre">Rachel Legendre</name><name sortKey="Pawlik, Alexandre" sort="Pawlik, Alexandre" uniqKey="Pawlik A" first="Alexandre" last="Pawlik">Alexandre Pawlik</name><name sortKey="Rodriguez Ord Ez, Maria Del Pilar" sort="Rodriguez Ord Ez, Maria Del Pilar" uniqKey="Rodriguez Ord Ez M" first="María Del Pilar" last="Rodríguez-Ord Ez">María Del Pilar Rodríguez-Ord Ez</name><name sortKey="Sismeiro, Odile" sort="Sismeiro, Odile" uniqKey="Sismeiro O" first="Odile" last="Sismeiro">Odile Sismeiro</name><name sortKey="Varet, Hugo" sort="Varet, Hugo" uniqKey="Varet H" first="Hugo" last="Varet">Hugo Varet</name><name sortKey="Varet, Hugo" sort="Varet, Hugo" uniqKey="Varet H" first="Hugo" last="Varet">Hugo Varet</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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