Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System
Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilt...
Gespeichert in:
| Veröffentlicht in: | Applied and Environmental Microbiology 2006-04, Vol.72 (4), p.2896-2904 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2904 |
|---|---|
| container_issue | 4 |
| container_start_page | 2896 |
| container_title | Applied and Environmental Microbiology |
| container_volume | 72 |
| creator | Tal, Yossi Watts, Joy E. M Schreier, Harold J |
| description | Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilters and the anaerobic sludge waste collection tank and was analyzed by amplifying 16S rRNA genes by PCR using anammox-selective and universal GC-clamped primers. Separation of amplified PCR products by denaturing gradient gel electrophoresis and sequencing of the different phylotypes revealed a diverse biofilter microbial community. While Planctomycetales were found in all three communities, the anaerobic denitrifying biofilters contained one clone that exhibited high levels of sequence similarity to known anammox bacteria. Fluorescence in situ hybridization studies using an anammox-specific probe confirmed the presence of anammox Planctomycetales in the microbial biofilm from the denitrifying biofilters, and anammox activity was observed in these biofilters, as detected by the ability to simultaneously consume ammonia and nitrite. To our knowledge, this is the first identification of anammox-related sequences in a marine recirculating aquaculture filtration system, and our findings provide a foundation for incorporating this important pathway for complete nitrogen removal in such systems. |
| doi_str_mv | 10.1128/AEM.72.4.2896-2904.2006 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_29795716</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>19974580</sourcerecordid><originalsourceid>FETCH-LOGICAL-c712t-855e84a7b3db9aaa4ae62b45922cb2cf235cf51d9a5bcb92c475ef1d715ed1d93</originalsourceid><addsrcrecordid>eNqNkttu1DAQhiMEokvhFahBAsFFFtvxIb5BSqsuILWqROm1NXGcXVc5tHZSdnkH3hlHu2KBm-XK45lv_rHGf5KcEDwnhOYfivPLuaRzNqe5EilVOEYYi0fJjGCVpzzLxONkhrFSKaUMHyXPQrjFGDMs8qfJERFcSaXELPlZdGB9XzqDirbtOze26dXaVe6H65boXazG7Po9OgUzWO8AQVehIoTeOBhsDM3gHtywQa5DC7e2VbpwTYtOXV-7JnYE1NcI0CV411n01RrnzdjAMKkX9yPEyzB6i643YbDt8-RJDU2wL3bncXKzOP929jm9uPr05ay4SI0kdEhzzm3OQJZZVSoAYGAFLRlXlJqSmppm3NScVAp4aUpFDZPc1qSShNsqprPj5ONW924sW1sZ2w0eGn3nXQt-o3tw-u9K51Z62T9owlge9xYF3u4EfH8_2jDo1gVjmwY6249BC5lnccP4IEgjxSUR_wEyqgRmB0GilGQ8PzyaMCm5zCbF1_-At_3ou_gBmmKuBFG5jJDcQsb3IXhb_94WwXrypI6e1JJqpidP6smTevJk7Hz557L3fTsTRuDNDoBgoKk9dMaFPSdllOHTO19tuZVbrr47bzWEVoNt92Mjc7Jlaug1LH3UubmmmGSYYCE5zbNfxXsA2w</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>205961987</pqid></control><display><type>article</type><title>Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System</title><source>American Society for Microbiology</source><source>MEDLINE</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Tal, Yossi ; Watts, Joy E. M ; Schreier, Harold J</creator><creatorcontrib>Tal, Yossi ; Watts, Joy E. M ; Schreier, Harold J</creatorcontrib><description>Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilters and the anaerobic sludge waste collection tank and was analyzed by amplifying 16S rRNA genes by PCR using anammox-selective and universal GC-clamped primers. Separation of amplified PCR products by denaturing gradient gel electrophoresis and sequencing of the different phylotypes revealed a diverse biofilter microbial community. While Planctomycetales were found in all three communities, the anaerobic denitrifying biofilters contained one clone that exhibited high levels of sequence similarity to known anammox bacteria. Fluorescence in situ hybridization studies using an anammox-specific probe confirmed the presence of anammox Planctomycetales in the microbial biofilm from the denitrifying biofilters, and anammox activity was observed in these biofilters, as detected by the ability to simultaneously consume ammonia and nitrite. To our knowledge, this is the first identification of anammox-related sequences in a marine recirculating aquaculture filtration system, and our findings provide a foundation for incorporating this important pathway for complete nitrogen removal in such systems.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/AEM.72.4.2896-2904.2006</identifier><identifier>PMID: 16597996</identifier><identifier>CODEN: AEMIDF</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Ammonia ; Ammonia - metabolism ; anaerobes ; Aquaculture ; Bacteria ; Bacteria, Anaerobic - classification ; Bacteria, Anaerobic - genetics ; Bacteria, Anaerobic - metabolism ; Biofilms - growth & development ; biofilters ; Biological and medical sciences ; biological treatment ; Biotechnology ; Deoxyribonucleic acid ; DNA ; DNA, Ribosomal - analysis ; Filtration - methods ; fish culture ; fixed-film biofilters ; Fundamental and applied biological sciences. Psychology ; genes ; In Situ Hybridization, Fluorescence ; mariculture ; Marine ; Microbial Ecology ; Microbiology ; Molecular Sequence Data ; Nitrates - metabolism ; Nitrosomonadaceae ; nucleotide sequences ; Organic waste treatment ; Oxidation ; Oxidation-Reduction ; Planctomycetales ; Polymorphism, Restriction Fragment Length ; recirculating aquaculture systems ; ribosomal RNA ; RNA, Ribosomal, 16S - genetics ; Seawater - microbiology ; Sequence Analysis, DNA ; wastewater treatment</subject><ispartof>Applied and Environmental Microbiology, 2006-04, Vol.72 (4), p.2896-2904</ispartof><rights>2006 INIST-CNRS</rights><rights>Copyright American Society for Microbiology Apr 2006</rights><rights>Copyright © 2006, American Society for Microbiology 2006</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c712t-855e84a7b3db9aaa4ae62b45922cb2cf235cf51d9a5bcb92c475ef1d715ed1d93</citedby><cites>FETCH-LOGICAL-c712t-855e84a7b3db9aaa4ae62b45922cb2cf235cf51d9a5bcb92c475ef1d715ed1d93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1448996/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1448996/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,3174,3175,27903,27904,53769,53771</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17700654$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16597996$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tal, Yossi</creatorcontrib><creatorcontrib>Watts, Joy E. M</creatorcontrib><creatorcontrib>Schreier, Harold J</creatorcontrib><title>Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System</title><title>Applied and Environmental Microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilters and the anaerobic sludge waste collection tank and was analyzed by amplifying 16S rRNA genes by PCR using anammox-selective and universal GC-clamped primers. Separation of amplified PCR products by denaturing gradient gel electrophoresis and sequencing of the different phylotypes revealed a diverse biofilter microbial community. While Planctomycetales were found in all three communities, the anaerobic denitrifying biofilters contained one clone that exhibited high levels of sequence similarity to known anammox bacteria. Fluorescence in situ hybridization studies using an anammox-specific probe confirmed the presence of anammox Planctomycetales in the microbial biofilm from the denitrifying biofilters, and anammox activity was observed in these biofilters, as detected by the ability to simultaneously consume ammonia and nitrite. To our knowledge, this is the first identification of anammox-related sequences in a marine recirculating aquaculture filtration system, and our findings provide a foundation for incorporating this important pathway for complete nitrogen removal in such systems.</description><subject>Ammonia</subject><subject>Ammonia - metabolism</subject><subject>anaerobes</subject><subject>Aquaculture</subject><subject>Bacteria</subject><subject>Bacteria, Anaerobic - classification</subject><subject>Bacteria, Anaerobic - genetics</subject><subject>Bacteria, Anaerobic - metabolism</subject><subject>Biofilms - growth & development</subject><subject>biofilters</subject><subject>Biological and medical sciences</subject><subject>biological treatment</subject><subject>Biotechnology</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Ribosomal - analysis</subject><subject>Filtration - methods</subject><subject>fish culture</subject><subject>fixed-film biofilters</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>genes</subject><subject>In Situ Hybridization, Fluorescence</subject><subject>mariculture</subject><subject>Marine</subject><subject>Microbial Ecology</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Nitrates - metabolism</subject><subject>Nitrosomonadaceae</subject><subject>nucleotide sequences</subject><subject>Organic waste treatment</subject><subject>Oxidation</subject><subject>Oxidation-Reduction</subject><subject>Planctomycetales</subject><subject>Polymorphism, Restriction Fragment Length</subject><subject>recirculating aquaculture systems</subject><subject>ribosomal RNA</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>Seawater - microbiology</subject><subject>Sequence Analysis, DNA</subject><subject>wastewater treatment</subject><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkttu1DAQhiMEokvhFahBAsFFFtvxIb5BSqsuILWqROm1NXGcXVc5tHZSdnkH3hlHu2KBm-XK45lv_rHGf5KcEDwnhOYfivPLuaRzNqe5EilVOEYYi0fJjGCVpzzLxONkhrFSKaUMHyXPQrjFGDMs8qfJERFcSaXELPlZdGB9XzqDirbtOze26dXaVe6H65boXazG7Po9OgUzWO8AQVehIoTeOBhsDM3gHtywQa5DC7e2VbpwTYtOXV-7JnYE1NcI0CV411n01RrnzdjAMKkX9yPEyzB6i643YbDt8-RJDU2wL3bncXKzOP929jm9uPr05ay4SI0kdEhzzm3OQJZZVSoAYGAFLRlXlJqSmppm3NScVAp4aUpFDZPc1qSShNsqprPj5ONW924sW1sZ2w0eGn3nXQt-o3tw-u9K51Z62T9owlge9xYF3u4EfH8_2jDo1gVjmwY6249BC5lnccP4IEgjxSUR_wEyqgRmB0GilGQ8PzyaMCm5zCbF1_-At_3ou_gBmmKuBFG5jJDcQsb3IXhb_94WwXrypI6e1JJqpidP6smTevJk7Hz557L3fTsTRuDNDoBgoKk9dMaFPSdllOHTO19tuZVbrr47bzWEVoNt92Mjc7Jlaug1LH3UubmmmGSYYCE5zbNfxXsA2w</recordid><startdate>20060401</startdate><enddate>20060401</enddate><creator>Tal, Yossi</creator><creator>Watts, Joy E. M</creator><creator>Schreier, Harold J</creator><general>American Society for Microbiology</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>7TV</scope><scope>F1W</scope><scope>H95</scope><scope>H98</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>7TB</scope><scope>KR7</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20060401</creationdate><title>Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System</title><author>Tal, Yossi ; Watts, Joy E. M ; Schreier, Harold J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c712t-855e84a7b3db9aaa4ae62b45922cb2cf235cf51d9a5bcb92c475ef1d715ed1d93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Ammonia</topic><topic>Ammonia - metabolism</topic><topic>anaerobes</topic><topic>Aquaculture</topic><topic>Bacteria</topic><topic>Bacteria, Anaerobic - classification</topic><topic>Bacteria, Anaerobic - genetics</topic><topic>Bacteria, Anaerobic - metabolism</topic><topic>Biofilms - growth & development</topic><topic>biofilters</topic><topic>Biological and medical sciences</topic><topic>biological treatment</topic><topic>Biotechnology</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Ribosomal - analysis</topic><topic>Filtration - methods</topic><topic>fish culture</topic><topic>fixed-film biofilters</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>genes</topic><topic>In Situ Hybridization, Fluorescence</topic><topic>mariculture</topic><topic>Marine</topic><topic>Microbial Ecology</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Nitrates - metabolism</topic><topic>Nitrosomonadaceae</topic><topic>nucleotide sequences</topic><topic>Organic waste treatment</topic><topic>Oxidation</topic><topic>Oxidation-Reduction</topic><topic>Planctomycetales</topic><topic>Polymorphism, Restriction Fragment Length</topic><topic>recirculating aquaculture systems</topic><topic>ribosomal RNA</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>Seawater - microbiology</topic><topic>Sequence Analysis, DNA</topic><topic>wastewater treatment</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tal, Yossi</creatorcontrib><creatorcontrib>Watts, Joy E. M</creatorcontrib><creatorcontrib>Schreier, Harold J</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>Pollution Abstracts</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Aquaculture Abstracts</collection><collection>ASFA: Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Civil Engineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and Environmental Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tal, Yossi</au><au>Watts, Joy E. M</au><au>Schreier, Harold J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System</atitle><jtitle>Applied and Environmental Microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>2006-04-01</date><risdate>2006</risdate><volume>72</volume><issue>4</issue><spage>2896</spage><epage>2904</epage><pages>2896-2904</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><coden>AEMIDF</coden><abstract>Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilters and the anaerobic sludge waste collection tank and was analyzed by amplifying 16S rRNA genes by PCR using anammox-selective and universal GC-clamped primers. Separation of amplified PCR products by denaturing gradient gel electrophoresis and sequencing of the different phylotypes revealed a diverse biofilter microbial community. While Planctomycetales were found in all three communities, the anaerobic denitrifying biofilters contained one clone that exhibited high levels of sequence similarity to known anammox bacteria. Fluorescence in situ hybridization studies using an anammox-specific probe confirmed the presence of anammox Planctomycetales in the microbial biofilm from the denitrifying biofilters, and anammox activity was observed in these biofilters, as detected by the ability to simultaneously consume ammonia and nitrite. To our knowledge, this is the first identification of anammox-related sequences in a marine recirculating aquaculture filtration system, and our findings provide a foundation for incorporating this important pathway for complete nitrogen removal in such systems.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>16597996</pmid><doi>10.1128/AEM.72.4.2896-2904.2006</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0099-2240 |
| ispartof | Applied and Environmental Microbiology, 2006-04, Vol.72 (4), p.2896-2904 |
| issn | 0099-2240 1098-5336 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_29795716 |
| source | American Society for Microbiology; MEDLINE; PubMed Central; Alma/SFX Local Collection |
| subjects | Ammonia Ammonia - metabolism anaerobes Aquaculture Bacteria Bacteria, Anaerobic - classification Bacteria, Anaerobic - genetics Bacteria, Anaerobic - metabolism Biofilms - growth & development biofilters Biological and medical sciences biological treatment Biotechnology Deoxyribonucleic acid DNA DNA, Ribosomal - analysis Filtration - methods fish culture fixed-film biofilters Fundamental and applied biological sciences. Psychology genes In Situ Hybridization, Fluorescence mariculture Marine Microbial Ecology Microbiology Molecular Sequence Data Nitrates - metabolism Nitrosomonadaceae nucleotide sequences Organic waste treatment Oxidation Oxidation-Reduction Planctomycetales Polymorphism, Restriction Fragment Length recirculating aquaculture systems ribosomal RNA RNA, Ribosomal, 16S - genetics Seawater - microbiology Sequence Analysis, DNA wastewater treatment |
| title | Anaerobic Ammonium-Oxidizing (Anammox) Bacteria and Associated Activity in Fixed-Film Biofilters of a Marine Recirculating Aquaculture System |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-26T05%3A38%3A21IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Anaerobic%20Ammonium-Oxidizing%20(Anammox)%20Bacteria%20and%20Associated%20Activity%20in%20Fixed-Film%20Biofilters%20of%20a%20Marine%20Recirculating%20Aquaculture%20System&rft.jtitle=Applied%20and%20Environmental%20Microbiology&rft.au=Tal,%20Yossi&rft.date=2006-04-01&rft.volume=72&rft.issue=4&rft.spage=2896&rft.epage=2904&rft.pages=2896-2904&rft.issn=0099-2240&rft.eissn=1098-5336&rft.coden=AEMIDF&rft_id=info:doi/10.1128/AEM.72.4.2896-2904.2006&rft_dat=%3Cproquest_pubme%3E19974580%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=205961987&rft_id=info:pmid/16597996&rfr_iscdi=true |