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  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher-id">87</journal-id>
      <journal-id journal-id-type="index">urn:lsid:arphahub.com:pub:A116C711-4C18-5A38-8F1E-5E97753A8A64</journal-id>
      <journal-title-group>
        <journal-title xml:lang="en">Folia Medica</journal-title>
        <abbrev-journal-title xml:lang="en">FM</abbrev-journal-title>
      </journal-title-group>
      <issn pub-type="ppub">0204-8043</issn>
      <issn pub-type="epub">1314-2143</issn>
      <publisher>
        <publisher-name>Plovdiv Medical University</publisher-name>
      </publisher>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="doi">10.3897/folmed.64.e64004</article-id>
      <article-id pub-id-type="publisher-id">64004</article-id>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Case Report</subject>
        </subj-group>
        <subj-group subj-group-type="scientific_subject">
          <subject>Dermatology</subject>
          <subject>Infectious diseases</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Green nail syndrome on the nail plate and bed related with <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> and <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> coinfection</article-title>
      </title-group>
      <contrib-group content-type="authors">
        <contrib contrib-type="author" corresp="yes">
          <name name-style="western">
            <surname>Ouzounova-Raykova</surname>
            <given-names>Vessela V.</given-names>
          </name>
          <email xlink:type="simple">pumpi@abv.bg</email>
          <xref ref-type="aff" rid="A1">1</xref>
        </contrib>
      </contrib-group>
      <aff id="A1">
        <label>1</label>
        <addr-line content-type="verbatim">Department of Medical Microbiology, Medical University of Sofia, Sofia, Bulgaria</addr-line>
        <institution>Medical University of Sofia</institution>
        <addr-line content-type="city">Sofia</addr-line>
        <country>Bulgaria</country>
      </aff>
      <author-notes>
        <fn fn-type="corresp">
          <p>Corresponding author: Vessela V. Ouzounova-Raykova, Department of Medical Microbiology, Medical University of Sofia, 2 Zdrave St., 1431 Sofia, Bulgaria; Email: <email xlink:type="simple">pumpi@abv.bg</email>; Tel.: +359 2 9172580</p>
        </fn>
      </author-notes>
      <pub-date pub-type="collection">
        <year>2022</year>
      </pub-date>
      <pub-date pub-type="epub">
        <day>30</day>
        <month>06</month>
        <year>2022</year>
      </pub-date>
      <volume>64</volume>
      <issue>3</issue>
      <fpage>547</fpage>
      <lpage>550</lpage>
      <uri content-type="arpha" xlink:href="http://openbiodiv.net/32B4E6AF-F62D-5878-B511-277E3147894A">32B4E6AF-F62D-5878-B511-277E3147894A</uri>
      <history>
        <date date-type="received">
          <day>05</day>
          <month>02</month>
          <year>2021</year>
        </date>
        <date date-type="accepted">
          <day>27</day>
          <month>03</month>
          <year>2021</year>
        </date>
      </history>
      <permissions>
        <copyright-statement>Vessela V. Ouzounova-Raykova</copyright-statement>
        <license license-type="creative-commons-attribution" xlink:href="http://creativecommons.org/licenses/by/4.0/" xlink:type="simple">
          <license-p>This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p>
        </license>
      </permissions>
      <abstract>
        <p>
          <bold>Abstract</bold>
        </p>
        <p>The green nail syndrome is characterized by discolouration of the nail plate frequently accompanied by chronic paronychia and onycholysis. The cause could be either bacterial or fungal infection.</p>
        <p>A 24-year-old woman presented to the clinic with a history of discolouration of the nail plate and bed accompanied by onycholysis. Scrapings from the nail were taken twice, microscopically examined, and cultured on media for isolation and identification of bacteria and <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name>. The first sample gave positive results for <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. After treatment and a lack of complete recovery of the nail, second sample was taken and subjected to the same tests. This time <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> infection was detected. Additional treatment was performed and the nail’s plate and bed were successfully treated.</p>
      </abstract>
      <kwd-group>
        <label>Keywords</label>
        <kwd>coinfection</kwd>
        <kwd><italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp.</kwd>
        <kwd><italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> spp.</kwd>
        <kwd>green nail syndrome</kwd>
        <kwd>skin appendage</kwd>
      </kwd-group>
    </article-meta>
  </front>
  <body>
    <sec sec-type="Introduction" id="SECID0E4E">
      <title>Introduction</title>
      <p>Green nail syndrome (GNS) is characterized by greenish discolouration of the nail plate (greenish-black, greenish-brown, greenish-yellow), frequently accompanied by chronic paronychia and onycholysis. The causes could be bacteria, <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> or a combination of these microorganisms in persons with predisposing factors such as trauma or consistent exposure of the skin to water, soaps or detergents.<sup>[<xref ref-type="bibr" rid="B1 B2 B3">1–3</xref>]</sup> We present a case of GNS in a young person developed some weeks after trauma of the finger nail.</p>
    </sec>
    <sec sec-type="CASE REPORT" id="SECID0EPF">
      <title>Case report</title>
      <p><italic>A 24-year-old woman presented with a history of greenish-yellow-brown discoloration of the fingernail plate accompanied by onycholysis developed a few weeks after a trauma. No paronychia was observed</italic><bold><italic>(Fig. <xref ref-type="fig" rid="F1">1</xref>)</italic></bold> . <italic>The history revealed no peculiarities related to the profession of the patient (she was a student), or to what her daily activities were. A written informed consent in accordance with the ethical standards of the Ethics Committee of Medical University of Sofia was taken. Nail scrapings were taken from the patient and subjected to microscopy (KOH preparation for direct mycological microscopy) and cultivated on blood agar, MacConkey agar, CHROMagar <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Candida">Candida</tp:taxon-name-part></tp:taxon-name> and SDA chloramphenicol (Becton Dickenson). Plates were incubated at 37°C for 24-48 hours for the routine microbiological testing and for additional three days at room temperature for <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> isolation.</italic></p>
      <fig id="F1" position="float" orientation="portrait">
        <object-id content-type="arpha">7BEB6A58-70FC-5136-92DD-BE150D0B9BC6</object-id>
        <label>Figure 1.</label>
        <caption>
          <p>Green nail syndrome with discolouration of the nail plate and bed accompanied by onycholysis.</p>
        </caption>
        <graphic xlink:href="foliamedica-64-3-e64004-g001.jpg" position="float" orientation="portrait" xlink:type="simple" id="oo_709085.jpg">
          <uri content-type="original_file">https://binary.pensoft.net/fig/709085</uri>
        </graphic>
      </fig>
      <p>
        <italic>The sample was positive for <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name> spp. and negative for <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name>. <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name> spp. was identified on the bases of colony morphology, Gram staining, catalase and oxidase test, bile-esculin test, and salt tolerance test (6.5% NaCl), biochemical reactions, as per standard microbiological guidelines. After unsuccessful combined oral and topical antibiotic treatment according to the strain sensitivity, new nail scrapping was taken and subjected to a new identical microbiological testing. In the second sample <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name> spp. infection was detected.</italic>
      </p>
    </sec>
    <sec sec-type="Discussion" id="SECID0EIH">
      <title>Discussion</title>
      <p>Infections of the nails can be caused by <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name>, bacteria, viruses or a combination of these.<sup>[<xref ref-type="bibr" rid="B4">4</xref>]</sup><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">Fungi</tp:taxon-name-part></tp:taxon-name>, the anthropophilic dermatophytes <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Trichophyton">Trichophyton</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="rubrum">rubrum</tp:taxon-name-part></tp:taxon-name></italic>, <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Epidermophyton">Epidermophyton</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="floccosum">floccosum</tp:taxon-name-part></tp:taxon-name></italic>, and <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Microsporum">Microsporum</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="gypseum">gypseum</tp:taxon-name-part></tp:taxon-name></italic> represent the main causative agents of onychomycoses. These are followed by the yeasts <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Candida">Candida</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="parapsilosis">parapsilosis</tp:taxon-name-part></tp:taxon-name></italic>, <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Candida">Candida</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="guilliermondii">guilliermondii</tp:taxon-name-part></tp:taxon-name></italic>, and <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Candida">Candida</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="albicans">albicans</tp:taxon-name-part></tp:taxon-name></italic>. <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> species are by far the most common cause of mold onychomycosis.<sup>[<xref ref-type="bibr" rid="B4">4</xref>]</sup></p>
      <p>Bacterial infections of the nails are not rare. Usually, Gram-negative bacteria are implicated in the process. <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Pseudomonas">Pseudomonas</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="aeruginosa">aeruginosa</tp:taxon-name-part></tp:taxon-name></italic> is the most frequently isolated bacteria. However, <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Klebsiella">Klebsiella</tp:taxon-name-part></tp:taxon-name></italic> spp. and Gram-positive bacteria such as <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Staphylococcus">Staphylococcus</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="aureus">aureus</tp:taxon-name-part></tp:taxon-name></italic> or <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Streptococcus">Streptococcus</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="pyogenes">pyogenes</tp:taxon-name-part></tp:taxon-name></italic> could be the protagonists in the infection.<sup>[<xref ref-type="bibr" rid="B5 B6 B7">5–7</xref>]</sup> In the discussed case, the 24-year-old woman presented with a greenish-yellow-brown discolouration affecting more than 80% of the surface area of the nail plate, accompanied by onycholysis developed a few weeks after trauma. Samples were collected and different microbiological tests were performed. Scrapings from the nail were subjected to microscopy study and the results were negative. The cultivation on a selective culture medium for isolation and identification of <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> did not show growing colonies. However, the cultures for bacteria showed positive results for <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp.</p>
      <p>It is known that <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. is a group of facultative anaerobic organisms that are part of the normal intestinal flora of humans and animals. They have been long recognized as important human pathogens and are becoming increasingly so. Important clinical infections caused by <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. include bacteremia, urinary tract infections, wound infections, endocarditis, meningitis, and diverticulitis. Enterococcal infections of the nails and their adjacent tissues are not common, but there are some data regarding this matter.<sup>[<xref ref-type="bibr" rid="B8">8</xref>]</sup></p>
      <p>The treatment of nail infections is challenging<sup>[<xref ref-type="bibr" rid="B9">9</xref>]</sup> and recommendations based on clinical trials or strict guidelines are lacking.<sup>[<xref ref-type="bibr" rid="B4">4</xref>]</sup> Therapy is based on the microorganism isolated, the number of nails affected, and the percentage of nail plate involvement. In the infectious nail diseases, local or systemic treatment could be applied. The topical therapy is preferred because it avoids the adverse effects over the metabolism.<sup>[<xref ref-type="bibr" rid="B10">10</xref>]</sup> Frequently, this therapy fails because of the poor penetration of the drug into the nail plate. The treatment usually requires an oral antifungal medication for several months owing to the higher concentration of therapeutic agents in the lesion.<sup>[<xref ref-type="bibr" rid="B11">11</xref>,12]</sup> So, mild infections affecting less than 50% of one or two nails could be subjected to topical medications, otherwise systemic therapy is recommended. We think that combined topical and oral treatment is probably the most effective regimen. Thus, the therapeutic scheme used for our patient was: amoxicillin/clavulanic acid 1 g BID for 10 days followed by tobramycin eye drops, 2-3 drops 3 times per day for 3 weeks. The medication was prescribed based on the susceptibility of the isolated strain. After two and a half months, the change in the nail structure was not satisfactory. The colour became whitish, the decreased sensitivity of the nail bed has recovered; however, the onycholysis could still be seen <bold>(Fig. <xref ref-type="fig" rid="F2">2</xref>)</bold>. The lack of desired clinical response made us conduct another microbiological study. This time <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> spp. infection was detected.</p>
      <fig id="F2" position="float" orientation="portrait">
        <object-id content-type="arpha">CDABEBD0-51C9-5223-B735-B95F28BA5DB1</object-id>
        <label>Figure 2.</label>
        <caption>
          <p>Green nail syndrome with some clinical response after 3 months specific treatment.</p>
        </caption>
        <graphic xlink:href="foliamedica-64-3-e64004-g002.jpg" position="float" orientation="portrait" xlink:type="simple" id="oo_709086.jpg">
          <uri content-type="original_file">https://binary.pensoft.net/fig/709086</uri>
        </graphic>
      </fig>
      <p><italic><tp:taxon-name>
            <tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part>
          </tp:taxon-name></italic> spp. are non-dermatophytic moulds. They are known as soil saprophytes, important plant pathogens as well as aetiological agents of opportunistic human infections and they are one of the three different types of <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> that could lead to the development of onychomycosis.<sup>[<xref ref-type="bibr" rid="B4">4</xref>,13]</sup></p>
      <p>In our opinion, the patient had sustained a bacterial and fungal coinfection. It is very likely that the inability to prove the mycotic agent resulted from the bacterial overgrowth in the culture, which suppressed the <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> development. When the bacteria were totally eliminated by the applied therapy, the nail remained with the fungal mono-infection alone. Meanwhile, a second sample was taken for a new microbiological test. The results for bacteria showed negative and <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> spp. was detected.</p>
      <p>The new therapeutic strategy consisted of topical treatment with an extempore prepared combination of reagents against <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> (thymolum, salicylic acid, resorcinol, acetic acid, iodine tincture, and alcohol 70°) applied twice per day for a sufficient amount of time. The nail plate was cured within 6 months after the initiation of this treatment.</p>
      <p>This is the first case in which <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. has been reported as a causative agent of GNS in coinfection with <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> spp. Although <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Pseudomonas">P.</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="aeruginosa">aeruginosa</tp:taxon-name-part></tp:taxon-name></italic>, known as the causative agent of GNS, was not detected in the sample, on the basis of the greenish-yellow-brown discolouration of the nail plate accompanied by onycholysis, and the detection of <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. with the possibility of some enterococci to synthesize yellowish pigment<sup>[<xref ref-type="bibr" rid="B14">14</xref>]</sup> give us the reason to assume that the described infection could be named as green nail syndrome.</p>
      <p>Bacterial and fungal coinfections of nails were described by Yang et al., too.<sup>[<xref ref-type="bibr" rid="B15">15</xref>]</sup> They found <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="solani">solani</tp:taxon-name-part></tp:taxon-name></italic> onychomycosis coinfected with <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Pseudomonas">P.</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="aeruginosa">aeruginosa</tp:taxon-name-part></tp:taxon-name></italic>. The researchers reported that coinfection with bacteria can prevent the isolation of the fungus because of bacterial overgrowth in culture which is consistent with our conclusions.</p>
      <p>It is worth noting that only about 50% of discoloured or dystrophic-appearing nails have an infection confirmed with microscope or culture. The differential diagnosis of GNS and other causes of onycholysis should include trauma (tight shoes, nail biting), eczema (irritant or allergic contact dermatitis), lichen planus, subungual melanoma, psoriatic nail disease, systemic diseases such as thyroid disease, diabetes, peripheral arterial disease, idiosyncratic drug reaction (especially tetracyclines, quinolones and psoralens) or chemical exposure to solutions containing pyocyanin or pyoverdine.</p>
    </sec>
    <sec sec-type="Conclusions" id="SECID0EEKAC">
      <title>Conclusions</title>
      <p>Green colouration of the nails accompanied by onycholysis should raise suspicion for GNS. Based on the practice in which <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name>, bacteria, and other microorganisms could affect the nail plate and bed, screening for co-infections is crucial. This is the first time in which <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Enterococcus">Enterococcus</tp:taxon-name-part></tp:taxon-name></italic> spp. has been reported as a causative agent of GNS in coinfection with <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Fusarium">Fusarium</tp:taxon-name-part></tp:taxon-name></italic> spp. The condition, however, was successfully treated <bold>(Fig. <xref ref-type="fig" rid="F3">3</xref>)</bold>.</p>
      <fig id="F3" position="float" orientation="portrait">
        <object-id content-type="arpha">C1F9B01A-EC5B-5739-A39D-5246DBD9D6E3</object-id>
        <label>Figure 3.</label>
        <caption>
          <p>Recovered nail plate and bed after 6 months treatment.</p>
        </caption>
        <graphic xlink:href="foliamedica-64-3-e64004-g003.jpg" position="float" orientation="portrait" xlink:type="simple" id="oo_709087.jpg">
          <uri content-type="original_file">https://binary.pensoft.net/fig/709087</uri>
        </graphic>
      </fig>
    </sec>
  </body>
  <back>
    <ref-list>
      <title>References</title>
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        <mixed-citation xlink:type="simple">4. Nenoff P, Paasch U, Handrick W. Infections of finger and toe nails due to <tp:taxon-name><tp:taxon-name-part taxon-name-part-type="kingdom">fungi</tp:taxon-name-part></tp:taxon-name> and bacteria. Hautarzt 2014; 65(4):337–48.</mixed-citation>
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</article>
