A novel approach to the antimicrobial activity of maggot debridement therapy
Identifieur interne : 001522 ( Istex/Corpus ); précédent : 001521; suivant : 001523A novel approach to the antimicrobial activity of maggot debridement therapy
Auteurs : Anders S. Andersen ; Dorthe Sandvang ; Kirk M. Schnorr ; Thomas Kruse ; Sren Neve ; Bo Joergensen ; Tonny Karlsmark ; Karen A. KrogfeltSource :
- Journal of Antimicrobial Chemotherapy [ 0305-7453 ] ; 2010-08.
Abstract
Objectives Commercially produced sterile green bottle fly Lucilia sericata maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments. Methods cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in Escherichia coli, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested in vitro against Gram-positive and Gram-negative bacterial strains. Results Lucifensin was active against Staphylococcus carnosus, Streptococcus pyogenes and Streptococcus pneumoniae (MIC 2 mg/L), as well as Staphylococcus aureus (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant S. aureus and glycopeptide-intermediate S. aureus isolates tested ranged from 8 to >128 mg/L. Conclusions The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.
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DOI: 10.1093/jac/dkq165
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Andersen</name><affiliation><mods:affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Sandvang, Dorthe" sort="Sandvang, Dorthe" uniqKey="Sandvang D" first="Dorthe" last="Sandvang">Dorthe Sandvang</name><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Schnorr, Kirk M" sort="Schnorr, Kirk M" uniqKey="Schnorr K" first="Kirk M." last="Schnorr">Kirk M. 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Krogfelt</name><affiliation><mods:affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: kak@ssi.dk</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:65996797900473D6EA7F9683995D11542FBE8D54</idno><date when="2010" year="2010">2010</date><idno type="doi">10.1093/jac/dkq165</idno><idno type="url">https://api.istex.fr/document/65996797900473D6EA7F9683995D11542FBE8D54/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">001522</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">A novel approach to the antimicrobial activity of maggot debridement therapy</title><author><name sortKey="Andersen, Anders S" sort="Andersen, Anders S" uniqKey="Andersen A" first="Anders S." last="Andersen">Anders S. Andersen</name><affiliation><mods:affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Sandvang, Dorthe" sort="Sandvang, Dorthe" uniqKey="Sandvang D" first="Dorthe" last="Sandvang">Dorthe Sandvang</name><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Schnorr, Kirk M" sort="Schnorr, Kirk M" uniqKey="Schnorr K" first="Kirk M." last="Schnorr">Kirk M. Schnorr</name><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Kruse, Thomas" sort="Kruse, Thomas" uniqKey="Kruse T" first="Thomas" last="Kruse">Thomas Kruse</name><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Neve, Sren" sort="Neve, Sren" uniqKey="Neve S" first="Sren" last="Neve">Sren Neve</name><affiliation><mods:affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Joergensen, Bo" sort="Joergensen, Bo" uniqKey="Joergensen B" first="Bo" last="Joergensen">Bo Joergensen</name><affiliation><mods:affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Karlsmark, Tonny" sort="Karlsmark, Tonny" uniqKey="Karlsmark T" first="Tonny" last="Karlsmark">Tonny Karlsmark</name><affiliation><mods:affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</mods:affiliation></affiliation></author><author><name sortKey="Krogfelt, Karen A" sort="Krogfelt, Karen A" uniqKey="Krogfelt K" first="Karen A." last="Krogfelt">Karen A. Krogfelt</name><affiliation><mods:affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: kak@ssi.dk</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Antimicrobial Chemotherapy</title><idno type="ISSN">0305-7453</idno><idno type="eISSN">1460-2091</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2010-08">2010-08</date><biblScope unit="volume">65</biblScope><biblScope unit="issue">8</biblScope><biblScope unit="page" from="1646">1646</biblScope><biblScope unit="page" to="1654">1654</biblScope></imprint><idno type="ISSN">0305-7453</idno></series><idno type="istex">65996797900473D6EA7F9683995D11542FBE8D54</idno><idno type="DOI">10.1093/jac/dkq165</idno><idno type="ArticleID">dkq165</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0305-7453</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract">Objectives Commercially produced sterile green bottle fly Lucilia sericata maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments. Methods cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in Escherichia coli, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested in vitro against Gram-positive and Gram-negative bacterial strains. Results Lucifensin was active against Staphylococcus carnosus, Streptococcus pyogenes and Streptococcus pneumoniae (MIC 2 mg/L), as well as Staphylococcus aureus (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant S. aureus and glycopeptide-intermediate S. aureus isolates tested ranged from 8 to >128 mg/L. Conclusions The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>Anders S. Andersen</name><affiliations><json:string>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</json:string><json:string>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</json:string><json:string>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</json:string></affiliations></json:item><json:item><name>Dorthe Sandvang</name><affiliations><json:string>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</json:string></affiliations></json:item><json:item><name>Kirk M. Schnorr</name><affiliations><json:string>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</json:string></affiliations></json:item><json:item><name>Thomas Kruse</name><affiliations><json:string>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</json:string></affiliations></json:item><json:item><name>Sren Neve</name><affiliations><json:string>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</json:string></affiliations></json:item><json:item><name>Bo Joergensen</name><affiliations><json:string>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</json:string></affiliations></json:item><json:item><name>Tonny Karlsmark</name><affiliations><json:string>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</json:string></affiliations></json:item><json:item><name>Karen A. Krogfelt</name><affiliations><json:string>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</json:string><json:string>E-mail: kak@ssi.dk</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>defensins</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Lucilia sericata</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>MRSA</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>antimicrobial peptides</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>wounds</value></json:item></subject><articleId><json:string>dkq165</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>Objectives Commercially produced sterile green bottle fly Lucilia sericata maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments. Methods cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in Escherichia coli, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested in vitro against Gram-positive and Gram-negative bacterial strains. Results Lucifensin was active against Staphylococcus carnosus, Streptococcus pyogenes and Streptococcus pneumoniae (MIC 2 mg/L), as well as Staphylococcus aureus (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant S. aureus and glycopeptide-intermediate S. aureus isolates tested ranged from 8 to >128 mg/L. Conclusions The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.</abstract><qualityIndicators><score>9</score><pdfVersion>1.5</pdfVersion><pdfPageSize>612.283 x 790.866 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>5</keywordCount><abstractCharCount>2102</abstractCharCount><pdfWordCount>6988</pdfWordCount><pdfCharCount>45153</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>277</abstractWordCount></qualityIndicators><title>A novel approach to the antimicrobial activity of maggot debridement therapy</title><genre><json:string>research-article</json:string></genre><host><volume>65</volume><publisherId><json:string>jac</json:string></publisherId><pages><last>1654</last><first>1646</first></pages><issn><json:string>0305-7453</json:string></issn><issue>8</issue><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><eissn><json:string>1460-2091</json:string></eissn><title>Journal of Antimicrobial Chemotherapy</title></host><categories><wos><json:string>INFECTIOUS DISEASES</json:string><json:string>PHARMACOLOGY & PHARMACY</json:string><json:string>MICROBIOLOGY</json:string></wos></categories><publicationDate>2010</publicationDate><copyrightDate>2010</copyrightDate><doi><json:string>10.1093/jac/dkq165</json:string></doi><id>65996797900473D6EA7F9683995D11542FBE8D54</id><score>0.22610493</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/65996797900473D6EA7F9683995D11542FBE8D54/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/65996797900473D6EA7F9683995D11542FBE8D54/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/65996797900473D6EA7F9683995D11542FBE8D54/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a">A novel approach to the antimicrobial activity of maggot debridement therapy</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Oxford University Press</publisher><availability><p>The Author 2010. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.</p></availability><date>2010-06-11</date></publicationStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">A novel approach to the antimicrobial activity of maggot debridement therapy</title><author xml:id="author-1"><persName><forename type="first">Anders S.</forename><surname>Andersen</surname></persName><affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</affiliation><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation></author><author xml:id="author-2"><persName><forename type="first">Dorthe</forename><surname>Sandvang</surname></persName><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation></author><author xml:id="author-3"><persName><forename type="first">Kirk M.</forename><surname>Schnorr</surname></persName><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation></author><author xml:id="author-4"><persName><forename type="first">Thomas</forename><surname>Kruse</surname></persName><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation></author><author xml:id="author-5"><persName><forename type="first">Sren</forename><surname>Neve</surname></persName><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation></author><author xml:id="author-6"><persName><forename type="first">Bo</forename><surname>Joergensen</surname></persName><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation></author><author xml:id="author-7"><persName><forename type="first">Tonny</forename><surname>Karlsmark</surname></persName><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation></author><author xml:id="author-8"><persName><forename type="first">Karen A.</forename><surname>Krogfelt</surname></persName><email>kak@ssi.dk</email><affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</affiliation></author></analytic><monogr><title level="j">Journal of Antimicrobial Chemotherapy</title><idno type="pISSN">0305-7453</idno><idno type="eISSN">1460-2091</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2010-08"></date><biblScope unit="volume">65</biblScope><biblScope unit="issue">8</biblScope><biblScope unit="page" from="1646">1646</biblScope><biblScope unit="page" to="1654">1654</biblScope></imprint></monogr><idno type="istex">65996797900473D6EA7F9683995D11542FBE8D54</idno><idno type="DOI">10.1093/jac/dkq165</idno><idno type="ArticleID">dkq165</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2010-06-11</date></creation><langUsage><language ident="en">en</language></langUsage><abstract><p>Objectives Commercially produced sterile green bottle fly Lucilia sericata maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments. Methods cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in Escherichia coli, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested in vitro against Gram-positive and Gram-negative bacterial strains. Results Lucifensin was active against Staphylococcus carnosus, Streptococcus pyogenes and Streptococcus pneumoniae (MIC 2 mg/L), as well as Staphylococcus aureus (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant S. aureus and glycopeptide-intermediate S. aureus isolates tested ranged from 8 to >128 mg/L. Conclusions The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.</p></abstract><textClass><keywords scheme="keyword"><list><head>keywords</head><item><term>defensins</term></item><item><term>Lucilia sericata</term></item><item><term>MRSA</term></item><item><term>antimicrobial peptides</term></item><item><term>wounds</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2010-06-11">Created</change><change when="2010-08">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/65996797900473D6EA7F9683995D11542FBE8D54/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article article-type="research-article"><front><journal-meta><journal-id journal-id-type="publisher-id">jac</journal-id><journal-id journal-id-type="hwp">jac</journal-id><journal-title>Journal of Antimicrobial Chemotherapy</journal-title><issn pub-type="ppub">0305-7453</issn><issn pub-type="epub">1460-2091</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.1093/jac/dkq165</article-id><article-id pub-id-type="publisher-id">dkq165</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original research</subject></subj-group></article-categories><title-group><article-title>A novel approach to the antimicrobial activity of maggot debridement therapy</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Andersen</surname><given-names>Anders S.</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af2">2</xref><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Sandvang</surname><given-names>Dorthe</given-names></name><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Schnorr</surname><given-names>Kirk M.</given-names></name><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Kruse</surname><given-names>Thomas</given-names></name><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Neve</surname><given-names>Søren</given-names></name><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Joergensen</surname><given-names>Bo</given-names></name><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Karlsmark</surname><given-names>Tonny</given-names></name><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Krogfelt</surname><given-names>Karen A.</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="af1"><label>1</label><addr-line>Department of Microbiological Surveillance and Research</addr-line>, <institution>Statens Serum Institut</institution>, <addr-line>Artillerivej 5, 2300 Copenhagen S</addr-line>, <country>Denmark</country></aff><aff id="af2"><label>2</label><institution>Novozymes A/S</institution>, <addr-line>Krogshoejvej 36, 2880 Bagsvaerd</addr-line>, <country>Denmark</country></aff><aff id="af3"><label>3</label><addr-line>Copenhagen Wound Healing Center</addr-line>, <institution>Bispebjerg Hospital</institution>, <addr-line>Bispebjerg bakke 23, 2400 Copenhagen NV</addr-line>, <country>Denmark</country></aff><author-notes><corresp id="cor1"><label>*</label>Corresponding author. Tel: <phone>+45-32-68-37-45</phone>; Fax: <fax>+45-32-68-31-47</fax>; E-mail: <email>kak@ssi.dk</email></corresp></author-notes><pub-date pub-type="ppub"><month>8</month><year>2010</year></pub-date><pub-date pub-type="epub"><day>11</day><month>6</month><year>2010</year></pub-date><volume>65</volume><issue>8</issue><fpage>1646</fpage><lpage>1654</lpage><history><date date-type="received"><day>20</day><month>11</month><year>2009</year></date><date date-type="rev-request"><day>28</day><month>1</month><year>2010</year></date><date date-type="rev-recd"><day>28</day><month>3</month><year>2010</year></date><date date-type="accepted"><day>21</day><month>4</month><year>2010</year></date></history><permissions><copyright-statement>© The Author 2010. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.</copyright-statement><copyright-year>2010</copyright-year><license license-type="creative-commons" xlink:href="http://creativecommons.org/licenses/by-nc/2.5/"><p>This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.</p></license></permissions><abstract><sec><title>Objectives</title><p>Commercially produced sterile green bottle fly <italic>Lucilia sericata</italic> maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments.</p></sec><sec><title>Methods</title><p>cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in <italic>Escherichia coli</italic>, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested <italic>in vitro</italic> against Gram-positive and Gram-negative bacterial strains.</p></sec><sec><title>Results</title><p>Lucifensin was active against <italic>Staphylococcus carnosus</italic>, <italic>Streptococcus pyogenes</italic> and <italic>Streptococcus pneumoniae</italic> (MIC 2 mg/L), as well as <italic>Staphylococcus aureus</italic> (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant <italic>S. aureus</italic> and glycopeptide-intermediate <italic>S. aureus</italic> isolates tested ranged from 8 to >128 mg/L.</p></sec><sec><title>Conclusions</title><p>The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.</p></sec></abstract><kwd-group><kwd>defensins</kwd><kwd><italic>Lucilia sericata</italic></kwd><kwd>MRSA</kwd><kwd>antimicrobial peptides</kwd><kwd>wounds</kwd></kwd-group></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>A novel approach to the antimicrobial activity of maggot debridement therapy</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>A novel approach to the antimicrobial activity of maggot debridement therapy</title></titleInfo><name type="personal"><namePart type="given">Anders S.</namePart><namePart type="family">Andersen</namePart><affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</affiliation><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Dorthe</namePart><namePart type="family">Sandvang</namePart><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Kirk M.</namePart><namePart type="family">Schnorr</namePart><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Thomas</namePart><namePart type="family">Kruse</namePart><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Sren</namePart><namePart type="family">Neve</namePart><affiliation>Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Bo</namePart><namePart type="family">Joergensen</namePart><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Tonny</namePart><namePart type="family">Karlsmark</namePart><affiliation>Copenhagen Wound Healing Center, Bispebjerg Hospital, Bispebjerg bakke 23, 2400 Copenhagen NV, Denmark</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Karen A.</namePart><namePart type="family">Krogfelt</namePart><affiliation>Department of Microbiological Surveillance and Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark</affiliation><affiliation>E-mail: kak@ssi.dk</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article"></genre><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">2010-08</dateIssued><dateCreated encoding="w3cdtf">2010-06-11</dateCreated><copyrightDate encoding="w3cdtf">2010</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract>Objectives Commercially produced sterile green bottle fly Lucilia sericata maggots are successfully employed by practitioners worldwide to clean a multitude of chronic necrotic wounds and reduce wound bacterial burdens during maggot debridement therapy (MDT). Secretions from the maggots exhibit antimicrobial activity along with other activities beneficial for wound healing. With the rise of multidrug-resistant bacteria, new approaches to identifying the active compounds responsible for the antimicrobial activity within this treatment are imperative. Therefore, the aim of this study was to use a novel approach to investigate the output of secreted proteins from the maggots under conditions mimicking clinical treatments. Methods cDNA libraries constructed from microdissected salivary glands and whole maggots, respectively, were treated with transposon-assisted signal trapping (TAST), a technique selecting for the identification of secreted proteins. Several putative secreted components of insect immunity were identified, including a defensin named lucifensin, which was produced recombinantly as a Trx-fusion protein in Escherichia coli, purified using immobilized metal affinity chromatography and reverse-phase HPLC, and tested in vitro against Gram-positive and Gram-negative bacterial strains. Results Lucifensin was active against Staphylococcus carnosus, Streptococcus pyogenes and Streptococcus pneumoniae (MIC 2 mg/L), as well as Staphylococcus aureus (MIC 16 mg/L). The peptide did not show antimicrobial activity towards Gram-negative bacteria. The MIC of lucifensin for the methicillin-resistant S. aureus and glycopeptide-intermediate S. aureus isolates tested ranged from 8 to >128 mg/L. Conclusions The TAST results did not reveal any highly secreted compounds with putative antimicrobial activity, implying an alternative antimicrobial activity of MDT. Lucifensin showed antimicrobial activities comparable to other defensins and could have potential as a future drug candidate scaffold, for redesign for other applications besides the topical treatment of infected wounds.</abstract><subject><genre>keywords</genre><topic>defensins</topic><topic>Lucilia sericata</topic><topic>MRSA</topic><topic>antimicrobial peptides</topic><topic>wounds</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Antimicrobial Chemotherapy</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0305-7453</identifier><identifier type="eISSN">1460-2091</identifier><identifier type="PublisherID">jac</identifier><identifier type="PublisherID-hwp">jac</identifier><part><date>2010</date><detail type="volume"><caption>vol.</caption><number>65</number></detail><detail type="issue"><caption>no.</caption><number>8</number></detail><extent unit="pages"><start>1646</start><end>1654</end></extent></part></relatedItem><identifier type="istex">65996797900473D6EA7F9683995D11542FBE8D54</identifier><identifier type="DOI">10.1093/jac/dkq165</identifier><identifier type="ArticleID">dkq165</identifier><accessCondition type="use and reproduction" contentType="copyright">The Author 2010. 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