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Proteins were quantified with Bradford assays (Bio-Rad) prior to the addition of trypsin, and then incubated overnight at 37ºC. The resulting peptides were dried and resuspended in 0.1% FA, and sonicated for 5 min prior to analysis. Individual EFA samples (n = 11) were used for the analysis of the protein content of this fluid. On the other hand a pooled sample (n = 8) was used for the analysis of the EFA natural peptide content. This sample was concentrated in an AMICON ultra centrifuge filter and the flow-through obtained was kept for analyzing the natural peptides. These were reduced with 10 mM DTT for 30 min, and alkylated with 25 mM IAA for another 30 min. Excess IAA was removed by an additional incubation with 10 mM DTT. Recovered peptides were desalted using stage-tip C18 microcolumns (Zip-tip, Millipore) and resuspended in 0.1% FA prior to MS analysis.PrideProteomics samples were loaded onto a nanoElute liquid chromatograph (Bruker) at 300 nl/min and ran twice using both a 60 and a 90 min linear gradient of 3–45% acetonitrile. Chromatograph was coupled on-line to a timsTOF Pro mass spectrometer using PASEF (Bruker Daltonics) where data was acquired. The mgf file generated by Data Analysis v3.0 (Bruker) was searched using Byonic software (Protein Metrics) through Proteome Discoverer v.1.4 software (Thermo). Applied search parameters were: 50 ppm precursor ion tolerance and 0.05 Da for fragment ions; Carbamidomethylation as fixed and methionine oxidation as variable modifications; up to two missed cleavages. Database search was performed against a UniProt/SwissProt database containing only Homo sapiens entries. Results were filtered using 1% FDR cutoff at protein level. Natural peptide separation was performed on a nanoACQUITY UPLC system (Waters) connected to either a Synapt G2si (Waters) or an LTQ Orbitrap XL ETD (Thermo) mass spectrometer. Samples were acquired as described before [21]. Searches were also performed using Byonic (Protein Metrics) on Proteome Discoverer v.1.4 software (Thermo). Orbi RAW files were directly searched, while mgf files were generated for Synapt files using ProteinLynx Global Server 3.0.2 (PLGS, Waters) and searched. Spectra were searched against a human UniProt/Swissprot database restricted to Homo sapiens using 10 ppm and 0.5Da for LTQ Orbitrap XL ETD searches and 15ppm and 0.2Da for the Synapt G2si searches. No enzyme specificity was defined. Carbamidomethylation was defined as fixed and methionine oxidation and N-terminal acetylation as variable modifications. Results were filtered using 1% FDR cutoff at protein level.ProteomicsFelix ElortzaProteomics Platform, CIC bioGUNE, CIBERehd, ProteoRed-ISCIII, Bizkaia Science and Technology Park, Derio, SpainPARTIALHomo Sapiens (human)mazkargorta@cicbiogune.es31958637 Azkargorta M, Bregón-Villahoz M, Escobes I, Ibáñez-Pérez J, Iloro I, Iglesias M, Diez-Zapirain M, Rabanal A, Prieto B, Moragues MD, Matorras R, Elortza F. In-depth proteomics and natural peptidomics analyses reveal antibacterial peptides in human endometrial fluid. J Proteomics. 2020 216:103652 10.1016/j.jprot.2020.103652Proteomics Platform CIC bioGUNESpainThe composition of endometrial fluid reflects the status of the endometrium; it is a good atraumatic source of information on embryo implantation processes and possible pathological conditions. Although some attempts have been made to characterise its proteome, the catalogue of its proteins remains incomplete and little has been done to analyse the natural peptides it contains. Here, we present a comprehensive analysis of the proteins and natural peptides of the endometrial fluid. The protein content of samples from 11 individuals was analysed using the novel timsTOF Pro mass spectrometer. We identified 4694 proteins with at least one peptide with FDR < 1%, of which 2261 were found in >50% of the samples. A pooled endometrial fluid sample was used for isolation and analysis of the natural peptides. Mass spectrometry analysis identified 3899 naturally occurring peptides from 238 different proteins. Among these, there were some putative natural antibacterial peptides. Antimicrobial activity of peptides derived from elafin and Cu/Zn superoxide dismutase was confirmed using microbiological assays. Our results substantially expand the catalogue of known endometrial fluid proteins and provide extensive new information on the natural peptide content of this fluid. SIGNIFICANCE: The endometrial fluid contains many proteins whose clinical relevance is still unknown. Some might be merely markers of endometrial function, but others might play a role in embryo nutrition and/or implantation. Human endometrial fluid analysis might open the door to new developments in embryo transfer strategies in in-vitro fertilisation programmes and lead to improvements in the composition of embryo culture media. Here, we report, for the first time, antimicrobial activity of endometrial fluid peptides. Such peptides could play an important role in the balance of the recently described uterine microbiota.In-depth proteomics and natural peptidomics analyses reveal antibacterial peptides in human endometrial fluid.Azkargorta Mikel M, Bregón-Villahoz Marta M, Escobes Iraide I, Ibáñez-Pérez Jone J, Iloro Ibon I, Iglesias Maria M, Diez-Zapirain Miren M, Rabanal Aintzane A, Prieto Begoña B, Moragues Maria-Dolores MD, Matorras Roberto R, Elortza Felix FNup32D, PLIP, MHC, determination, GPATCH6, rad, CSMF, protein, 2-(indol-3-yl)ethanoic acid, CG15720, fluid, composed of, Type 2A-interacting protein, peptide, Myo, Polypeptides, peptido, IAA, dJ69E11.3, B1, Min, protein aggregate, DmelCG42273, HTATIP, nup154, DmelCG3533, rsh, peptides, Myo2, HAND, hypoplasia, min, chemical analysis., mAPC, composition, bHLHa28, DROMHC, Protein Degradation, foot, hand, AI047805, NMMII, NOR-1, cPLA2, sample, D1, NMM, stage, TIP, Dmel_CG4346, Myo II, forefoot of quadruped, symmetric dyschromatosis of the extremities, Degradations, CHN, ZC2HC5, Dmel_CG7826, Mhc-c[1], HTATIP1, tip, ANAC091, myoII, Putative MAPK-activating protein PM10, (Indol-3-yl)acetate, paw, Dm1, Digestion, DmelCG42628, Raps, IES, TIP60, Dmel_CG7835, Mnb, MNB, DLK, ribosome-associated degradation, content, Protein Digestions, familial reticulate acropigmentation of Dohi, dyschromatosis symmetrica hereditaria 1, Dmel_CG15720, pooled, LNKN-1, AW124434, study protocol, beta Trypsin, KIAA1847, REM3, Osi, heteroauxin, fore paw, CG3533, Polypeptide, MyoII, forefoot, tlp, RAD1, TIP41, ZIP, Zip, developmental stage, Degradation, rapsyn, Gene, Myo-II HC, protein-containing complex, DmelCG5692, CG7826, ESA1, zip, method, Dm nmII, reduced, method used in an experiment, Arabidopsis NAC domain containing protein 91, Gene Products, CG7835, CG18144, CG42273, zk, tiny, CG15792, ZIPK, l(2)10432, ZC3HDC9, AI573420, pins, Myo-II, zip/MyoII, dHand, beta-Trypsin, Indoleacetic acid, Protein Degradations, PINS, Dhand, Solution, DmelCG4579, ms(2)zk, hand-C, (indol-3-yl)acetic acid, dHAND, peptidos, l(2)01501, DmnmII, D8Wsu49e, ZIP3, PTHB1, l(2)02957, small, GPATC6, NOR1, DYRK1, Nor1, CG42628, CG5692, ZC3H9, protein complex, BcDNA:LD21772, Proteins, E(br), DSH, compositionality, Protein Digestion, Digestions, Peptide, polypeptide, native protein, Proteolyses, nmMHC, C18, fore-paw, Protein, chemical analysis, Mhc-c, Dyrk1, forefeet, Pins, DmelCG15792, nzip, AU020952, nup32D, PRSS, MINOR, underdeveloped, DmelCG18144, CC1, Tripcellim, 3-Indolylessigsaeure, CG4579, CDA08, sample population, ME-IV, plan specification, Protein Gene Products, Hands, Gene Proteins, Trypure, fore foot, structure, CG4346, TCV-interacting protein, forepaw, Dronm-MII, anon-WO0140519.37, TEC, Peptid, RAD, Rad, assay, Minor, zpr, nmy-2Human, fluid., human being, Man (Taxonomy), Homo sapiens, Peptidomics, Man, Modern Man, human, ModernBru, MeCN, NCMe, ion, Raw, Somatomedin-C, protein, contrasted, ran, peptide, Polypeptides, L-Isomer Methionine, ranGap, ranGAP, Methionine, peptido, CH3-C#N, Plgs, Min, Software Engineering, Analysis, AAF30287, protein aggregate, Computer Program, DmelCG42273, DmelCG9999, Svc, rangap, Man (Taxonomy), peptides, Analyses, M, Open, SD, min, SF, Computer Programs and Programming, mAPC, Kitl, Mast cell growth factor, AI047805, ppm, Mechano growth factor, Sd, Soluble KIT ligand, ran-1, Sl, Racemethionine, IGF-I, CT28175, Modern, acetylation, 2-Amino-4-(methylthio)butyric acid, Dmel_CG7826, IBP1, not genetically inherited, Source Softwares, results, sKITLG, xran, Software Tools, Programs, Program, Computer Applications, Dm1, Ionen, Computer Applications Software, Computer Applications Softwares, Methionin, Softwares, Hmet, Igf-1, fixed, Dmel_CG7835, Rasl2-8, Mnb, MNB, parent ion, SHEP7, Software Applications, mast cell growth factor, Source Software, Stem cell factor, AW124434, STAT5, dRanGAP, human, Kitlg, data analysis, Applications, KL-1, 1110030K05Rik, precursor ion, Biocatalyst, CG9999, Polypeptide, methyl cyanide, Liquimeth, Gb, Computer Software Applications, Clo, 2-amino-4-(methylsulfanyl)butanoic acid, human being, Peptidomics, Slf, SLF, Biocatalysts, L Isomer, dran, Gene, FPH2, Computer, precursor, protein-containing complex, CG7826, ara24, RanGAP, Human, Sd-RanGAP, Sd-RanGap, Homo sapiens, DmelCG1404, CG7835, Gene Products, CG42273, L-Methionine, Del(8)44H, Man, Pedameth, Application, Open Source Softwares, anatomical systems, Software Application, iones, Open Source Software, Acetylations, ions, man, Computer Software Application, Con, Enzyme, data processing, 2-amino-4-(methylthio)butanoic acid, Tools, 10^[-6], blz, peptidos, liquid, metionina, Applications Software, Open Source, DYRK1, Computer Software, 1-(14)C-labeled, 3H-labeled, DL-Methionine, protein complex, AI447866, Proteins, ran10A, acetonitrile, L-Isomer, gsp1, Peptide, OK/SW-cl.81, CG1404, Tool, polypeptide, TC4, tc4, Software Tool, native protein, Protein, Dyrk1, Software, ACETONITRILE, cyanomethane, Met, Data Base, AU020952, l(1)G0075, Col4a-1, ARA24, Protein., RanGap1, CC1, Engineering, ethanenitrile, ME-IV, Protein Gene Products, Computer Programs, Gene Proteins, Applications Softwares, Ion, Data, MGF, Mgf, Modern Man, c-Kit ligand, Gsp1, alpha-amino-gamma-methylmercaptobutyric acid, Peptid, SCF, variable, Data Analyses, Scl, RanEndometria, human being, Procedures, determination, Blastocyst Implantation, extraembryonic placenta, FEZL, Gene, protein, Spectrum Analyses, fluid, protein-containing complex, composed of, Human, Blastocyst Implantations, peptide, Polypeptides, method, Techniques, peptido, Homo sapiens, TOF, Method, method used in an experiment, Gene Products, Mass, Studies, Analysis, protein aggregate, Man, Technique, ZNF312, Mass Spectroscopy, ASD2, uterine mucosa, Mass Spectrum Analysis, Ovum Implantations, Man (Taxonomy), peptides, Analyses, TACHD, maternal part of placenta, composition, Study, endometrium, uterine endometrium, Methodological Studies, uterine mucous membrane, Implantation, sample, peptidos, tunica mucosa (endometrium), Nidations, FEZ, Decidual Cell Reaction, maternal decidual layer, protein complex, Embryo Implantations, Modern, Proteins, decidua basalis, incomplete, Nidation, maternal placenta, compositionality, Procedure, Spectrum Analysis, Peptide, uterine decidua, Spectroscopy, abolished, polypeptide, Implantations, MS, Blastocyst, native protein, placenta maternal decidual layer, Protein, chemical analysis, blastocyst implantation, Mass Spectrum Analyses, Embryo, VSD1, Mass Spectrum, Decidual Cell Reactions, Ovum Implantation, Protein., content, Spectrometry, extraembryonic part of placenta, pooled, Methodological, Methodological Study, human, sample population, Protein Gene Products, plan specification, Gene Proteins, ZFP312, Modern Man, structure, Peptid, assay, Polypeptide, Ovum, tunica mucosa uteriGhrfr, Skin derived, Prenatal Structures, Activity, determination, antimicrobials, plant germ, extraembryonic placenta, Cobalt, Mn-Superoxide Dismutase, Mn-Superoxide, antimicrobial agents, Skin Derived Antileukoproteinase, protein, Cu-Superoxide Dismutase, Progress Reports, fluid, Long Term, Blastocyst Implantations, peptide, Polypeptides, microbicides, peptido, Roles, Summary Report, Concepts, Fe-Zn Superoxide, Cobalt Superoxide, protein aggregate, Transfer, Effect, Cu-Superoxide, Summary Reports, antimicrobial, Ovum Implantations, Man (Taxonomy), peptides, Progress Report, Antibiotika, Microbial Community Structure, maternal part of placenta, Fe-Zn Superoxide Dismutase, Clinical Importance, purification, Source of Information, isolation and purification, Progress, Field Reports, Fe Superoxide Dismutase, sample, Role Concepts, Microbial Community Structures, tunica mucosa (endometrium), Cu Superoxide Dismutase, Long-Term Effects, Tubal Embryo Transfer, antibiotic, Presumptive, Iron Superoxide, Decidual Cell Reaction, maternal decidual layer, anatomical protrusion, Dismutase, embryo, Modern, decidua basalis, Longterm Effect, lit, Manganese Superoxide Dismutase, developing organism, maternal placenta, Microbial, Significance, Implantations, Data Sources, Blastocyst, Role Concept, Tubal Embryo Stage Transfer, Investigative Report, Playthings and Play, placenta maternal decidual layer, Role, Plaything, microbicide, Ag Zn Superoxide Dismutase, Microbial Communities, little, Decidual Cell Reactions, Ovum Implantation, Iron, Microbial Community Compositions, Antileukoproteinase, Protease Inhibitor WAP3, Relevance, Field, Toys, Fe-Superoxide Dismutase, Mn-SOD, Prenatal Structure, human, Composition, Report, Skin-derived, spine, Superoxide, Manganese Superoxide, Polypeptide, Ovum, Proteomes, Human Microbiomes, Community, Manganese, Endometria, Microbial Community Composition, Plays, Cobalt Superoxide Dismutase, human being, antibiotique, Mn SOD, Fe-Zn, Effects, Blastocyst Implantation, Importance, Prenatal, Gene, Protease, protein-containing complex, protrusion, Skin-Derived Antileukoproteinase, Human, Inhibitor WAP3, Blastocyst Transfer, Investigative, Homo sapiens, isolation, Embryo Transfers, Gene Products, Fe Zn Superoxide Dismutase, Clinical Significance, Man, antibiotics, Fe-Superoxide, Ag-Zn Superoxide, Toy, uterine mucosa, Clinical, Playthings, Longterm, Hemocuprein, Long-Term, Structure, Puppets, embryonic organism, endometrium, uterine endometrium, uterine mucous membrane, Implantation, Presumptive Structure, Information Source, Embryonic Structure, Play, peptidos, Long-Term Effect, Investigative Reports, Nidations, Puppet, Human Microbiome, Antibiotikum, Superoxide Dismutase, Erythrocuprein, Microbiome., Community Composition, Skin-Derived, protein complex, Embryo Implantations, Proteins, total expressed protein, Nidation, function, Peptide, Culture, uterine decidua, Concept, Iron Superoxide Dismutase, polypeptide, native protein, Media, Ag-Zn Superoxide Dismutase, Embryos, Protein, chemical analysis, Research Reports, Long Term Effects, blastocyst implantation, Microbiome, Embryonic, Embryo, Microbiomes, Microbial Community, Community Structure, extraembryonic part of placenta, sample population, Longterm Effects, Presumptive Structures, Protein Gene Products, Gene Proteins, developmental tissue, Superoxide:superoxide oxidoreductase, Reports, Protease Inhibitor 3, Peptidase Inhibitor 3, Modern Man, Sources of Information, GRO:0005343, Microbiotas, Peptid, Data Source, assay, Ag-Zn, Transfers, Summary, General activity, tunica mucosa uteri, Field Report, Mn Superoxide DismutaseHuman, Polypeptides, human being, peptido, Man (Taxonomy), peptides, Homo sapiens, Peptidomics, Modern Man, Modern, fluid., Peptid, peptidos, Polypeptide, Man, Peptide, humanfalseIn-depth proteomics and natural peptidomics analyses of human endometrial fluidEndometrial fluid reflects the status of the endometrium, being a good atraumatic source of information on embryo implantation and some pathological conditions. Although some attempts have been made to characterize it, the catalogue of the human endometrial fluid proteins remains incomplete and little has been done regarding the analysis of the containing natural peptides. In this work, we have thoroughly analyzed the protein and natural peptide content of endometrial fluid. Protein content coming from 11 different individuals was analyzed using the novel TIMS TOF Pro mass spectrometer. A total of 4694 proteins were identified with at least one peptide with FDR<1%, of which 2261 were identified in more than 50% of the samples. Besides, a pooled endometrial fluid sample was used for isolating the natural peptides and analyzed by mass spectrometry. Following this method we identified 3899 naturally-occurring peptides from 238 different proteins.2020-09-292019-12-11PXD016707NEWT:3555NEWT:71647NEWT:330879NEWT:35554NCBITaxon:12939NEWT:32046NEWT:544496NEWT:59529NEWT:2042546NEWT:32049NEWT:1392696NEWT:259447NEWT:295546NCBITaxon:2719036NEWT:45351NEWT:445974NEWT:43179NEWT:180454NEWT:5722NCBITaxon:1280NEWT:112503NEWT:1129NEWT:309807NEWT:10245NEWT:55153NEWT:89184NEWT:309800NEWT:281395NEWT:1211601NEWT:876138NEWT:44271NEWT:193516NEWT:351581NEWT:43186NEWT:428146NEWT:111205NEWT:498257NEWT:10036NEWT:1590NEWT:498019NEWT:1351NEWT:1438992NEWT:1352NEWT:2649997NEWT:1147161NEWT:661410NEWT:638632NEWT:263737NEWT:224326NEWT:1333499NEWT:376619NCBITaxon:79857NEWT:1096976NEWT:5702NEWT:95648NEWT:1589NEWT:135622NEWT:1349NEWT:35786NEWT:1580NEWT:399784NEWT:96731NEWT:383379NEWT:1401257NEWT:418106NEWT:10029NEWT:913645NEWT:44491NEWT:641809NEWT:27933NEWT:556484NEWT:317447NEWT:4688NEWT:7955NEWT:111225NEWT:7719NEWT:7959NEWT:2261NEWT:868565NEWT:31156NEWT:3112NEWT:79329NEWT:4442NEWT:192875NEWT:1214915NEWT:12637NEWT:59729NEWT:2164133NEWT:284218NEWT:295105NEWT:108061NEWT:60711NEWT:1316931NEWT:224308NEWT:3347NEWT:511145NEWT:160621NEWT:212790NEWT:931281NEWT:4432NEWT:5762NEWT:5763NEWT:658457NEWT:1310161NEWT:77133NEWT:295358NEWT:145481NEWT:29058NCBITaxon:79824NEWT:2246NEWT:2652724NEWT:1912919NEWT:5757NEWT:68415NCBITaxon:4563NEWT:5755NEWT:44688NEWT:44689NEWT:44447NEWT:752555NEWT:498211NEWT:5759NEWT:347256NEWT:1736231NEWT:5518NEWT:398007NEWT:1527468NEWT:1392NEWT:498217NEWT:498216NEWT:2242NEWT:11320NEWT:286NEWT:391619NEWT:246196NEWT:287NEWT:246197NEWT:1367269NEWT:145458NEWT:633149NEWT:1149786NEWT:10239NEWT:68895NEWT:44685NEWT:161934NEWT:4897NEWT:1148NEWT:5744NEWT:72901NEWT:213863NEWT:5508NEWT:3329NEWT:5507NEWT:410661NEWT:55571NEWT:35500NEWT:1235816NEWT:1140NCBITaxon:2157NEWT:1143NEWT:4896NEWT:1287689NEWT:1390NEWT:11557NEWT:1094343NEWT:1462472NEWT:1336795NEWT:644042NEWT:294NEWT:1773NEWT:1182590NEWT:3712NEWT:82380NEWT:3711NEWT:935293NEWT:375146NEWT:270643NEWT:2065263NEWT:177437NEWT:1772NEWT:10418NEWT:118698NEWT:1616117NEWT:263NEWT:118696NEWT:34865NEWT:52283NEWT:11988NEWT:284812NEWT:8175NEWT:400667NEWT:43330NEWT:1603293NEWT:408169NEWT:44664NEWT:47946NEWT:459758NEWT:3702NEWT:1245466NEWT:243277NEWT:1246791NEWT:7067NEWT:118694NEWT:586722NEWT:2850NEWT:118691NEWT:34871NEWT:33548NEWT:274NEWT:7070NCBITaxon:287NEWT:408172NEWT:408170NEWT:96794NEWT:3708NEWT:332648NEWT:44670NEWT:536231NEWT:376219NEWT:219813NEWT:1510NEWT:1436733NEWT:460519NEWT:1247411NEWT:1515NEWT:572307NEWT:1432138NEWT:1424507NEWT:1194599NEWT:272844NEWT:9483NEWT:333760NEWT:56636NEWT:1513458NEWT:1233681NEWT:2709072NCBITaxon:40559NEWT:506599NEWT:2853422NEWT:81077NEWT:84588NEWT:1679718NEWT:480NEWT:65349NEWT:67767NEWT:46835NEWT:109757NEWT:582580NEWT:300852NEWT:294607NEWT:1502NEWT:128017NEWT:376686NEWT:95486NEWT:508771NEWT:1883446NEWT:253NCBITaxon:2759NEWT:1233435NEWT:93061NEWT:93062NEWT:34613NCBITaxon:4932NEWT:109760NEWT:29031NEWT:943274NEWT:644223NEWT:235443NEWT:108458NEWT:5936NEWT:272623NEWT:272624NEWT:320637NEWT:886559NEWT:3750NEWT:55521NEWT:983964NEWT:118499NEWT:11706NEWT:32644NEWT:527796NEWT:1130798NEWT:55529NEWT:499175NEWT:109779NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:476272NEWT:3747NEWT:195051NEWT:3988NEWT:1116234NEWT:561007NEWT:1255228NEWT:649908NEWT:410289NEWT:373153NEWT:3983NEWT:203696NEWT:352472NEWT:97441NEWT:1071661NEWT:1202532NEWT:360094NEWT:1408223NEWT:470NEWT:1408224NEWT:41364NEWT:5911NEWT:1313NEWT:1218097NEWT:1080348NCBITaxon:50557NEWT:1314NEWT:39251NEWT:39491NEWT:212142NCBITaxon:5811NEWT:29491NEWT:101841NEWT:153481NEWT:2014887NEWT:33952NEWT:153009NEWT:261756NEWT:571256NEWT:40483NEWT:63366NEWT:63367NEWT:688333NEWT:215402NCBITaxon:548681NEWT:318586NCBITaxon:5820NEWT:1547NEWT:272634NEWT:198107NEWT:9031NEWT:27292NEWT:1872122NEWT:1308NEWT:7091NEWT:1034015NEWT:760568NEWT:141262NEWT:108931NEWT:1293497NEWT:1055524NEWT:4087NEWT:9778NEWT:306NEWT:150475NEWT:303NEWT:267872NEWT:97477NEWT:7111NEWT:347515NEWT:172269NEWT:9534NEWT:6269NEWT:5180NEWT:256737NEWT:4090NEWT:4092NEWT:9541NEWT:8694NEWT:4093NEWT:33936NEWT:4096NEWT:8692NEWT:2903NEWT:185579NEWT:941442NEWT:536088NEWT:13076NEWT:1226408NEWT:7108NEWT:40479NEWT:43989NEWT:4076NEWT:142615NEWT:317NEWT:1006581NEWT:33940NEWT:885318NEWT:39488NEWT:550NEWT:4081NEWT:273068NEWT:554NEWT:98334NEWT:451516NEWT:552NEWT:4084NEWT:1325291NEWT:63577NEWT:36185NEWT:1225786NEWT:7574NEWT:1715256NEWT:30640NEWT:575412NEWT:929793NEWT:1507179NEWT:29204NEWT:28112NEWT:114155NEWT:6494NEWT:6491NEWT:507601NEWT:520NEWT:186441NEWT:634955NEWT:643680NEWT:214092NCBITaxon:6157NEWT:6239NEWT:13095NEWT:162425NEWT:216257NEWT:102169NEWT:208893NEWT:9986NEWT:4054NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:8655NEWT:5147NEWT:98351NEWT:1263854NEWT:118503NEWT:2059687NEWT:160488NEWT:28104NEWT:207559NEWT:407821NCBITaxon:2NEWT:85057NEWT:985076NEWT:568708NEWT:1215323NEWT:986NEWT:52641NEWT:7159NEWT:267671NEWT:28532NEWT:7160NEWT:353152NEWT:8496NEWT:7165NEWT:1257118NEWT:27448NEWT:40674NEWT:1194669NEWT:51329NEWT:235612NEWT:243230NEWT:443906NEWT:1080772NEWT:519NEWT:8479NEWT:8485NEWT:6063NEWT:49240NEWT:1328388NEWT:104395NEWT:667127NEWT:9557NEW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