Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil)
Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosp...
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| creator | Lima, Sandrine M. A. Melo, Janaína. G. S. Militão, Gardênia C. G. Lima, Gláucia M. S. do Carmo A. Lima, Maria Aguiar, Jaciana S. Araújo, Renata M. Braz-Filho, Raimundo Marchand, Pascal Araújo, Janete M. Silva, Teresinha G. |
| description | Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of
Paullinia cupana
. The Actinobacteria was identified as
Streptomyces hygroscopicus
ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with
n
-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C
54
H
88
O
18
) based on
1
H and
13
C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC
50
of 1 μg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC
50
of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and |
| doi_str_mv | 10.1007/s00253-016-7886-9 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_hal_p</sourceid><recordid>TN_cdi_hal_primary_oai_HAL_hal_03825272v1</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A476743001</galeid><sourcerecordid>A476743001</sourcerecordid><originalsourceid>FETCH-LOGICAL-c538t-bcfeb93475b587d68ef66ddd251bb068ca5775e873a4baf02b0e01ef03243aad3</originalsourceid><addsrcrecordid>eNp1kstu1DAUhiMEotPCA7BBkdi0EgFf4kuWYQRMpUFITFlbjuNMXCVxsB3EzHv0fXHI0AISK-vY37n8Pn-SvIDgDQSAvfUAIIIzAGnGOKdZ8ShZwRyjDFCYP05WADKSMVLws-Tc-1sAIOKUPk3OEGOEEcBXyd26lU6qoJ05ymDskNomDa1OK2NVq3ujZPc6HduDN7az-yWUQ532ujbKDLJLR2dH7YLRfs7dBafHYPuDinF72DvrlR2Nmnxarm8-7bJikxpvOxl0nTbO9r-6lb08xt6X75w8mu7qWfKkkZ3Xz0_nRfL1w_ub9Sbbfv54vS63mSKYh6xSja4KnDNSEc5qynVDaV3XiMCqApQrSaJQzRmWeSUbgCqgAdQNwCjHUtb4Irla6rayE6MzvXQHYaURm3Ir5juAOSKIoe8wspcLG_V-m7QPojde6a6Tg7aTF5BTjjFjnEb01T_orZ1c_KuZIhQUFDPyQO1lp4UZGhviJuaioswZZTmOC3sY8UQpOwT9I-zl5L243n35m4ULq-Kve6ebe00QiNkxYnGMiI4Rs2NEEXNenmadqrjT-4zfFokAWgAfn4a9dn-I-W_Vnwnoypk</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1856096375</pqid></control><display><type>article</type><title>Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil)</title><source>MEDLINE</source><source>SpringerLink Journals</source><creator>Lima, Sandrine M. A. ; Melo, Janaína. G. S. ; Militão, Gardênia C. G. ; Lima, Gláucia M. S. ; do Carmo A. Lima, Maria ; Aguiar, Jaciana S. ; Araújo, Renata M. ; Braz-Filho, Raimundo ; Marchand, Pascal ; Araújo, Janete M. ; Silva, Teresinha G.</creator><creatorcontrib>Lima, Sandrine M. A. ; Melo, Janaína. G. S. ; Militão, Gardênia C. G. ; Lima, Gláucia M. S. ; do Carmo A. Lima, Maria ; Aguiar, Jaciana S. ; Araújo, Renata M. ; Braz-Filho, Raimundo ; Marchand, Pascal ; Araújo, Janete M. ; Silva, Teresinha G.</creatorcontrib><description>Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of
Paullinia cupana
. The Actinobacteria was identified as
Streptomyces hygroscopicus
ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with
n
-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C
54
H
88
O
18
) based on
1
H and
13
C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC
50
of 1 μg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC
50
of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and <3.9 μg/mL, respectively). In conclusion, the EB1 fraction and elaiophylin of
S. hygroscopicus
have potent antimicrobial, antifungal, and anticancer activities.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-016-7886-9</identifier><identifier>PMID: 27757508</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Actinobacteria ; Analysis ; Anti-Infective Agents - chemistry ; Anti-Infective Agents - isolation & purification ; Antibiotics ; Antimicrobial agents ; Antineoplastic Agents - chemistry ; Antineoplastic Agents - isolation & purification ; Applied Microbial and Cell Physiology ; Bacteria ; Bacteria - drug effects ; Biological Products - chemistry ; Biological Products - isolation & purification ; Biomass ; Biomedical and Life Sciences ; Biotechnology ; Brazil ; Cancer ; Cell Line, Tumor ; Cell Survival - drug effects ; Cytotoxicity ; Drug resistance ; Fermentation ; Fungi - drug effects ; Glycerol ; Health aspects ; Humans ; Inhibitory Concentration 50 ; Life Sciences ; Metabolites ; Microbial Genetics and Genomics ; Microbial Sensitivity Tests ; Microbiology ; Microorganisms ; Natural products ; Paullinia - growth & development ; Paullinia cupana ; Pharmaceutical industry ; Physiology ; Rhizosphere ; Soil Microbiology ; Spectrum Analysis ; Streptomyces ; Streptomyces - chemistry ; Streptomyces - isolation & purification ; Streptomyces - metabolism ; Streptomyces - physiology ; Streptomyces hygroscopicus ; Studies ; Yeast</subject><ispartof>Applied microbiology and biotechnology, 2017, Vol.101 (2), p.711-723</ispartof><rights>Springer-Verlag Berlin Heidelberg 2016</rights><rights>COPYRIGHT 2017 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, 2017.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c538t-bcfeb93475b587d68ef66ddd251bb068ca5775e873a4baf02b0e01ef03243aad3</citedby><cites>FETCH-LOGICAL-c538t-bcfeb93475b587d68ef66ddd251bb068ca5775e873a4baf02b0e01ef03243aad3</cites><orcidid>0000-0001-7773-8642</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-016-7886-9$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-016-7886-9$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27757508$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-03825272$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Lima, Sandrine M. A.</creatorcontrib><creatorcontrib>Melo, Janaína. G. S.</creatorcontrib><creatorcontrib>Militão, Gardênia C. G.</creatorcontrib><creatorcontrib>Lima, Gláucia M. S.</creatorcontrib><creatorcontrib>do Carmo A. Lima, Maria</creatorcontrib><creatorcontrib>Aguiar, Jaciana S.</creatorcontrib><creatorcontrib>Araújo, Renata M.</creatorcontrib><creatorcontrib>Braz-Filho, Raimundo</creatorcontrib><creatorcontrib>Marchand, Pascal</creatorcontrib><creatorcontrib>Araújo, Janete M.</creatorcontrib><creatorcontrib>Silva, Teresinha G.</creatorcontrib><title>Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil)</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of
Paullinia cupana
. The Actinobacteria was identified as
Streptomyces hygroscopicus
ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with
n
-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C
54
H
88
O
18
) based on
1
H and
13
C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC
50
of 1 μg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC
50
of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and <3.9 μg/mL, respectively). In conclusion, the EB1 fraction and elaiophylin of
S. hygroscopicus
have potent antimicrobial, antifungal, and anticancer activities.</description><subject>Actinobacteria</subject><subject>Analysis</subject><subject>Anti-Infective Agents - chemistry</subject><subject>Anti-Infective Agents - isolation & purification</subject><subject>Antibiotics</subject><subject>Antimicrobial agents</subject><subject>Antineoplastic Agents - chemistry</subject><subject>Antineoplastic Agents - isolation & purification</subject><subject>Applied Microbial and Cell Physiology</subject><subject>Bacteria</subject><subject>Bacteria - drug effects</subject><subject>Biological Products - chemistry</subject><subject>Biological Products - isolation & purification</subject><subject>Biomass</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Brazil</subject><subject>Cancer</subject><subject>Cell Line, Tumor</subject><subject>Cell Survival - drug effects</subject><subject>Cytotoxicity</subject><subject>Drug resistance</subject><subject>Fermentation</subject><subject>Fungi - drug effects</subject><subject>Glycerol</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Inhibitory Concentration 50</subject><subject>Life Sciences</subject><subject>Metabolites</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbial Sensitivity Tests</subject><subject>Microbiology</subject><subject>Microorganisms</subject><subject>Natural products</subject><subject>Paullinia - growth & development</subject><subject>Paullinia cupana</subject><subject>Pharmaceutical industry</subject><subject>Physiology</subject><subject>Rhizosphere</subject><subject>Soil Microbiology</subject><subject>Spectrum Analysis</subject><subject>Streptomyces</subject><subject>Streptomyces - chemistry</subject><subject>Streptomyces - isolation & purification</subject><subject>Streptomyces - metabolism</subject><subject>Streptomyces - physiology</subject><subject>Streptomyces hygroscopicus</subject><subject>Studies</subject><subject>Yeast</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp1kstu1DAUhiMEotPCA7BBkdi0EgFf4kuWYQRMpUFITFlbjuNMXCVxsB3EzHv0fXHI0AISK-vY37n8Pn-SvIDgDQSAvfUAIIIzAGnGOKdZ8ShZwRyjDFCYP05WADKSMVLws-Tc-1sAIOKUPk3OEGOEEcBXyd26lU6qoJ05ymDskNomDa1OK2NVq3ujZPc6HduDN7az-yWUQ532ujbKDLJLR2dH7YLRfs7dBafHYPuDinF72DvrlR2Nmnxarm8-7bJikxpvOxl0nTbO9r-6lb08xt6X75w8mu7qWfKkkZ3Xz0_nRfL1w_ub9Sbbfv54vS63mSKYh6xSja4KnDNSEc5qynVDaV3XiMCqApQrSaJQzRmWeSUbgCqgAdQNwCjHUtb4Irla6rayE6MzvXQHYaURm3Ir5juAOSKIoe8wspcLG_V-m7QPojde6a6Tg7aTF5BTjjFjnEb01T_orZ1c_KuZIhQUFDPyQO1lp4UZGhviJuaioswZZTmOC3sY8UQpOwT9I-zl5L243n35m4ULq-Kve6ebe00QiNkxYnGMiI4Rs2NEEXNenmadqrjT-4zfFokAWgAfn4a9dn-I-W_Vnwnoypk</recordid><startdate>2017</startdate><enddate>2017</enddate><creator>Lima, Sandrine M. 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A. ; Melo, Janaína. G. S. ; Militão, Gardênia C. G. ; Lima, Gláucia M. S. ; do Carmo A. Lima, Maria ; Aguiar, Jaciana S. ; Araújo, Renata M. ; Braz-Filho, Raimundo ; Marchand, Pascal ; Araújo, Janete M. ; Silva, Teresinha G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c538t-bcfeb93475b587d68ef66ddd251bb068ca5775e873a4baf02b0e01ef03243aad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Actinobacteria</topic><topic>Analysis</topic><topic>Anti-Infective Agents - chemistry</topic><topic>Anti-Infective Agents - isolation & purification</topic><topic>Antibiotics</topic><topic>Antimicrobial agents</topic><topic>Antineoplastic Agents - chemistry</topic><topic>Antineoplastic Agents - isolation & purification</topic><topic>Applied Microbial and Cell Physiology</topic><topic>Bacteria</topic><topic>Bacteria - drug effects</topic><topic>Biological Products - chemistry</topic><topic>Biological Products - isolation & purification</topic><topic>Biomass</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Brazil</topic><topic>Cancer</topic><topic>Cell Line, Tumor</topic><topic>Cell Survival - drug effects</topic><topic>Cytotoxicity</topic><topic>Drug resistance</topic><topic>Fermentation</topic><topic>Fungi - drug effects</topic><topic>Glycerol</topic><topic>Health aspects</topic><topic>Humans</topic><topic>Inhibitory Concentration 50</topic><topic>Life Sciences</topic><topic>Metabolites</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbial Sensitivity Tests</topic><topic>Microbiology</topic><topic>Microorganisms</topic><topic>Natural products</topic><topic>Paullinia - growth & development</topic><topic>Paullinia cupana</topic><topic>Pharmaceutical industry</topic><topic>Physiology</topic><topic>Rhizosphere</topic><topic>Soil Microbiology</topic><topic>Spectrum Analysis</topic><topic>Streptomyces</topic><topic>Streptomyces - chemistry</topic><topic>Streptomyces - isolation & purification</topic><topic>Streptomyces - metabolism</topic><topic>Streptomyces - physiology</topic><topic>Streptomyces hygroscopicus</topic><topic>Studies</topic><topic>Yeast</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lima, Sandrine M. 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A.</au><au>Melo, Janaína. G. S.</au><au>Militão, Gardênia C. G.</au><au>Lima, Gláucia M. S.</au><au>do Carmo A. Lima, Maria</au><au>Aguiar, Jaciana S.</au><au>Araújo, Renata M.</au><au>Braz-Filho, Raimundo</au><au>Marchand, Pascal</au><au>Araújo, Janete M.</au><au>Silva, Teresinha G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil)</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2017</date><risdate>2017</risdate><volume>101</volume><issue>2</issue><spage>711</spage><epage>723</epage><pages>711-723</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of
Paullinia cupana
. The Actinobacteria was identified as
Streptomyces hygroscopicus
ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with
n
-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C
54
H
88
O
18
) based on
1
H and
13
C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC
50
of 1 μg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC
50
of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and <3.9 μg/mL, respectively). In conclusion, the EB1 fraction and elaiophylin of
S. hygroscopicus
have potent antimicrobial, antifungal, and anticancer activities.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>27757508</pmid><doi>10.1007/s00253-016-7886-9</doi><tpages>13</tpages><orcidid>https://orcid.org/0000-0001-7773-8642</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2017, Vol.101 (2), p.711-723 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_hal_primary_oai_HAL_hal_03825272v1 |
| source | MEDLINE; SpringerLink Journals |
| subjects | Actinobacteria Analysis Anti-Infective Agents - chemistry Anti-Infective Agents - isolation & purification Antibiotics Antimicrobial agents Antineoplastic Agents - chemistry Antineoplastic Agents - isolation & purification Applied Microbial and Cell Physiology Bacteria Bacteria - drug effects Biological Products - chemistry Biological Products - isolation & purification Biomass Biomedical and Life Sciences Biotechnology Brazil Cancer Cell Line, Tumor Cell Survival - drug effects Cytotoxicity Drug resistance Fermentation Fungi - drug effects Glycerol Health aspects Humans Inhibitory Concentration 50 Life Sciences Metabolites Microbial Genetics and Genomics Microbial Sensitivity Tests Microbiology Microorganisms Natural products Paullinia - growth & development Paullinia cupana Pharmaceutical industry Physiology Rhizosphere Soil Microbiology Spectrum Analysis Streptomyces Streptomyces - chemistry Streptomyces - isolation & purification Streptomyces - metabolism Streptomyces - physiology Streptomyces hygroscopicus Studies Yeast |
| title | Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil) |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-15T09%3A01%3A29IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_hal_p&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Characterization%20of%20the%20biochemical,%20physiological,%20and%20medicinal%20properties%20of%20Streptomyces%20hygroscopicus%20ACTMS-9H%20isolated%20from%20the%20Amazon%20(Brazil)&rft.jtitle=Applied%20microbiology%20and%20biotechnology&rft.au=Lima,%20Sandrine%20M.%20A.&rft.date=2017&rft.volume=101&rft.issue=2&rft.spage=711&rft.epage=723&rft.pages=711-723&rft.issn=0175-7598&rft.eissn=1432-0614&rft_id=info:doi/10.1007/s00253-016-7886-9&rft_dat=%3Cgale_hal_p%3EA476743001%3C/gale_hal_p%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1856096375&rft_id=info:pmid/27757508&rft_galeid=A476743001&rfr_iscdi=true |