A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro
Aims To use a flow‐based method to establish, quantify and visualize biofilms of Ureaplasma parvum. Methods and Results Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour chan...
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| Veröffentlicht in: | Journal of applied microbiology 2021-11, Vol.131 (5), p.2579-2585 |
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| container_title | Journal of applied microbiology |
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| creator | Rowlands, R.S. Kragh, K. Sahu, S. Maddocks, S.E. Bolhuis, A. Spiller, O.B. Beeton, M.L. |
| description | Aims
To use a flow‐based method to establish, quantify and visualize biofilms of Ureaplasma parvum.
Methods and Results
Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow‐cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High‐resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation.
Conclusions
Flow is an essential requirement for the establishment of U. parvum biofilms.
Significance and Impact of the Study
This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence‐associated with adhesion. |
| doi_str_mv | 10.1111/jam.15120 |
| format | Article |
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To use a flow‐based method to establish, quantify and visualize biofilms of Ureaplasma parvum.
Methods and Results
Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow‐cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High‐resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation.
Conclusions
Flow is an essential requirement for the establishment of U. parvum biofilms.
Significance and Impact of the Study
This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence‐associated with adhesion.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/jam.15120</identifier><identifier>PMID: 33899996</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Antiinfectives and antibacterials ; Antimicrobial agents ; biofilm ; Biofilms ; Biomass ; Cell culture ; flow cell ; Flow velocity ; quantification ; Scanning electron microscopy ; Ureaplasma parvum ; Virulence</subject><ispartof>Journal of applied microbiology, 2021-11, Vol.131 (5), p.2579-2585</ispartof><rights>2021 The Authors. published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.</rights><rights>2021 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.</rights><rights>2021. This article is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3880-815cb9672c9c853cc906a1deceb7c6e6c7b8444ed3eecad8d69881352defbad3</citedby><cites>FETCH-LOGICAL-c3880-815cb9672c9c853cc906a1deceb7c6e6c7b8444ed3eecad8d69881352defbad3</cites><orcidid>0000-0002-6292-0772</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjam.15120$$EPDF$$P50$$Gwiley$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjam.15120$$EHTML$$P50$$Gwiley$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,1416,27923,27924,45573,45574</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33899996$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rowlands, R.S.</creatorcontrib><creatorcontrib>Kragh, K.</creatorcontrib><creatorcontrib>Sahu, S.</creatorcontrib><creatorcontrib>Maddocks, S.E.</creatorcontrib><creatorcontrib>Bolhuis, A.</creatorcontrib><creatorcontrib>Spiller, O.B.</creatorcontrib><creatorcontrib>Beeton, M.L.</creatorcontrib><title>A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro</title><title>Journal of applied microbiology</title><addtitle>J Appl Microbiol</addtitle><description>Aims
To use a flow‐based method to establish, quantify and visualize biofilms of Ureaplasma parvum.
Methods and Results
Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow‐cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High‐resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation.
Conclusions
Flow is an essential requirement for the establishment of U. parvum biofilms.
Significance and Impact of the Study
This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence‐associated with adhesion.</description><subject>Antiinfectives and antibacterials</subject><subject>Antimicrobial agents</subject><subject>biofilm</subject><subject>Biofilms</subject><subject>Biomass</subject><subject>Cell culture</subject><subject>flow cell</subject><subject>Flow velocity</subject><subject>quantification</subject><subject>Scanning electron microscopy</subject><subject>Ureaplasma parvum</subject><subject>Virulence</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><recordid>eNp10D1PwzAQBmALgShfA38AWWKBIa0dx44zVhWfKmIpYowc5yJSOXGwk1b995i2MCBxy93w6NXpReiSkjENM1mqZkw5jckBOqFM8CgWaXy4vZOIkzQeoVPvl4RQRrg4RiPGZBZGnKD3KXbwOdQOGmh7XFmHK2PXuLcYWlUYwP0H4BJWYGy3JbbCbw5UZ5RvFO6UWw0NLmpb1abxuG7xqu6dPUdHlTIeLvb7DC3u7xazx2j--vA0m84jzaQkkaRcF1n4VmdacqZ1RoSiJWgoUi1A6LSQSZJAyQC0KmUpMikp43EJVaFKdoZudrGds58D-D5vaq_BGNWCHXwecypTliWpCPT6D13awbXhuaBkwmKeJTyo253SznrvoMo7VzfKbXJK8u-y81B2vi072Kt94lA0UP7Kn3YDmOzAujaw-T8pf56-7CK_AIZliSw</recordid><startdate>202111</startdate><enddate>202111</enddate><creator>Rowlands, R.S.</creator><creator>Kragh, K.</creator><creator>Sahu, S.</creator><creator>Maddocks, S.E.</creator><creator>Bolhuis, A.</creator><creator>Spiller, O.B.</creator><creator>Beeton, M.L.</creator><general>Oxford University Press</general><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TM</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-6292-0772</orcidid></search><sort><creationdate>202111</creationdate><title>A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro</title><author>Rowlands, R.S. ; Kragh, K. ; Sahu, S. ; Maddocks, S.E. ; Bolhuis, A. ; Spiller, O.B. ; Beeton, M.L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3880-815cb9672c9c853cc906a1deceb7c6e6c7b8444ed3eecad8d69881352defbad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Antiinfectives and antibacterials</topic><topic>Antimicrobial agents</topic><topic>biofilm</topic><topic>Biofilms</topic><topic>Biomass</topic><topic>Cell culture</topic><topic>flow cell</topic><topic>Flow velocity</topic><topic>quantification</topic><topic>Scanning electron microscopy</topic><topic>Ureaplasma parvum</topic><topic>Virulence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rowlands, R.S.</creatorcontrib><creatorcontrib>Kragh, K.</creatorcontrib><creatorcontrib>Sahu, S.</creatorcontrib><creatorcontrib>Maddocks, S.E.</creatorcontrib><creatorcontrib>Bolhuis, A.</creatorcontrib><creatorcontrib>Spiller, O.B.</creatorcontrib><creatorcontrib>Beeton, M.L.</creatorcontrib><collection>Wiley-Blackwell Open Access Titles</collection><collection>Wiley Free Content</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rowlands, R.S.</au><au>Kragh, K.</au><au>Sahu, S.</au><au>Maddocks, S.E.</au><au>Bolhuis, A.</au><au>Spiller, O.B.</au><au>Beeton, M.L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro</atitle><jtitle>Journal of applied microbiology</jtitle><addtitle>J Appl Microbiol</addtitle><date>2021-11</date><risdate>2021</risdate><volume>131</volume><issue>5</issue><spage>2579</spage><epage>2585</epage><pages>2579-2585</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><abstract>Aims
To use a flow‐based method to establish, quantify and visualize biofilms of Ureaplasma parvum.
Methods and Results
Absorbance readings of a U. parvum HPA5 culture were taken at 550 nm every 3 h for 30 h in order to establish a growth curve, with viability determined by the number of colour changing units (CCUs). Biofilms were established using the DTU flow‐cell with a flow rate of 0·01 ml min−1 and compared to the static control. Titres of bacteria were determined by CCU and biofilm biomass was quantified by Syto9 staining and COMSTAT analysis. High‐resolution images were obtained by scanning electron microscopy (SEM). Flow resulted in significantly more biofilm and higher cell titre (0·599 µm3/µm2 ± 0·152 and 4 × 108 CCU per ml, respectively) compared with static conditions (0·008 µm3/µm2 ± 0·010 and no recoverable cells, respectively). SEM revealed pleomorphic cells, with signs of budding and possible membrane vesicle formation.
Conclusions
Flow is an essential requirement for the establishment of U. parvum biofilms.
Significance and Impact of the Study
This is the first quantification of biofilm biomass formed by U. parvum. It is now possible to establish viable biofilms of U. parvum which will allow for future testing of antimicrobial agents and understanding of virulence‐associated with adhesion.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>33899996</pmid><doi>10.1111/jam.15120</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0002-6292-0772</orcidid><oa>free_for_read</oa></addata></record> |
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| source | Oxford University Press Journals All Titles (1996-Current); Wiley Online Library All Journals |
| subjects | Antiinfectives and antibacterials Antimicrobial agents biofilm Biofilms Biomass Cell culture flow cell Flow velocity quantification Scanning electron microscopy Ureaplasma parvum Virulence |
| title | A requirement for flow to enable the development of Ureaplasma parvum biofilms in vitro |
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