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<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Mar. Sci.</journal-id>
<journal-title-group>
<journal-title>Frontiers in Marine Science</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Mar. Sci.</abbrev-journal-title>
</journal-title-group>
<issn pub-type="epub">2296-7745</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
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<article-meta>
<article-id pub-id-type="doi">10.3389/fmars.2025.1663783</article-id>
<article-version article-version-type="Version of Record" vocab="NISO-RP-8-2008"/>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Original Research</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>The trapping of microplastics in the <italic>Posidonia oceanica</italic> aegagropiles in Tunisian coastal areas&#x2014;Southern Mediterranean</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" corresp="yes">
<name><surname>Sghaier</surname><given-names>Dhouha Belhaj</given-names></name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="corresp" rid="c001"><sup>*</sup></xref>
<xref ref-type="author-notes" rid="fn003"><sup>&#x2020;</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/381176/overview"/>
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<contrib contrib-type="author">
<name><surname>Chniti</surname><given-names>Ines</given-names></name>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
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<contrib contrib-type="author">
<name><surname>Barhoumi-Slimi</surname><given-names>Thouraya</given-names></name>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
<xref ref-type="aff" rid="aff3"><sup>3</sup></xref>
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<contrib contrib-type="author">
<name><surname>Zaaboub</surname><given-names>Noureddine</given-names></name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
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<contrib contrib-type="author">
<name><surname>EL Bour</surname><given-names>Monia</given-names></name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
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<aff id="aff1"><label>1</label><institution>National Institute of Marine Sciences and Technologies (INSTM), University of Carthage</institution>, <city>Tunis</city>,&#xa0;<country country="tn">Tunisia</country></aff>
<aff id="aff2"><label>2</label><institution>High Institute of Environmental Science and Technology, Technopark of Borj Cedria, University of Carthage</institution>, <city>Tunis</city>,&#xa0;<country country="tn">Tunisia</country></aff>
<aff id="aff3"><label>3</label><institution>Department of Chemistry, Laboratory of Structural (Bio)Organic Chemistry and Polymers, Faculty of Sciences of Tunis, University of Tunis El Manar</institution>, <city>Tunis</city>,&#xa0;<country country="tn">Tunisia</country></aff>
<author-notes>
<corresp id="c001"><label>*</label>Correspondence: Dhouha Belhaj Sghaier, <email xlink:href="mailto:dhouhasghaier@hotmail.fr">dhouhasghaier@hotmail.fr</email></corresp>
<fn fn-type="other" id="fn003">
<label>&#x2020;</label>
<p>ORCID: Dhouha Belhaj Sghaier, <uri xlink:href="https://orcid.org/0000-0001-5733-8377">orcid.org/0000-0001-5733-8377</uri></p></fn>
</author-notes>
<pub-date publication-format="electronic" date-type="pub" iso-8601-date="2025-10-14">
<day>14</day>
<month>10</month>
<year>2025</year>
</pub-date>
<pub-date publication-format="electronic" date-type="collection">
<year>2025</year>
</pub-date>
<volume>12</volume>
<elocation-id>1663783</elocation-id>
<history>
<date date-type="received">
<day>10</day>
<month>07</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>17</day>
<month>09</month>
<year>2025</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2025 Sghaier, Chniti, Barhoumi-Slimi, Zaaboub and EL Bour.</copyright-statement>
<copyright-year>2025</copyright-year>
<copyright-holder>Sghaier, Chniti, Barhoumi-Slimi, Zaaboub and EL Bour</copyright-holder>
<license>
<ali:license_ref start_date="2025-10-14">https://creativecommons.org/licenses/by/4.0/</ali:license_ref>
<license-p>This is an open-access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License (CC BY)</ext-link>. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p>
</license>
</permissions>
<abstract>
<p>Plastic and microplastic debris (MP) constitute the most important pollutants of the solid litter with a high risk of sediment accumulation. <italic>Posidonia oceanica</italic> (L.) Delile is the main marine seagrass of the Mediterranean Sea which forms immense underwater meadows and deposits of seagrass beds covering the facades of sandy beaches. They are formed by roots and rhizome fragments gathered in fibrous marine balls, called aegagropiles (EGs), having the ability to trap several pollutants from the beaches and mainly microplastic. The present study aims at evaluating microplastic contamination in aegagropiles collected from four locations along the Tunisian coast in the southern Mediterranean basin (two northern sites (S1 and S2) and two southern-central sites (S3 and S4). Microscopic analysis revealed that red and blue microplastics dominated at all sites, with black fibers and fragments being the most prevalent forms and yellow (S3) and transparent particles in S1, S2, and S4. Polymer identification conducted using nuclear magnetic resonance (NMR) and Fourier-transform infrared spectroscopy (FTIR) detected microplastic types with contamination levels and microplastic accumulation variation among the four sites including polystyrene (PS) at sites S1, S3, and S4; ethyl vinyl acetate (EVA) at S1 and S3; polyethylene terephthalate (PET) at S1 and S2; and polyvinyl chloride (PVC) at S2 and S3. Our results highlight close relationships between anthropogenic activities, extensive plastic use, and elevated microplastic pollution in marine ecosystems, particularly in seagrass beds. These findings emphasize the importance of monitoring microplastic contamination to preserve the health of Mediterranean coastal environments.</p>
</abstract>
<kwd-group>
<kwd><italic>Posidonia oceanica</italic></kwd>
<kwd>aegagropile</kwd>
<kwd>microplastics</kwd>
<kwd>acid digestion</kwd>
<kwd>NMR spectroscopy</kwd>
<kwd>FTIR</kwd>
<kwd>stereomicroscopy</kwd>
</kwd-group>
<funding-group>
<funding-statement>The author(s) declare that no financial support was received for the research and/or publication of this article.</funding-statement>
</funding-group>
<counts>
<fig-count count="5"/>
<table-count count="2"/>
<equation-count count="0"/>
<ref-count count="78"/>
<page-count count="12"/>
<word-count count="4941"/>
</counts>
<custom-meta-group>
<custom-meta>
<meta-name>section-in-acceptance</meta-name>
<meta-value>Marine Pollution</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
</front>
<body>
<sec id="s1" sec-type="intro">
<title>Introduction</title>
<p>Waste management is becoming a major issue with the increase in human population density; indeed, recent studies have highlighted a serious issue of marine litter (<xref ref-type="bibr" rid="B17">De Luca et&#xa0;al., 2025</xref>; <xref ref-type="bibr" rid="B60">Sarkar et&#xa0;al., 2025</xref>). It is well documented that the human-produced waste which accumulates in marine environments consists essentially of large quantities of microplastics in water like rivers, lakes, seas and oceans (<xref ref-type="bibr" rid="B20">Fraissinet et&#xa0;al., 2025</xref>; <xref ref-type="bibr" rid="B25">Gurjar et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B31">Jamsek et&#xa0;al., 2024</xref>). Research has demonstrated that UV light and low temperature facilitate the breakdown of conventional plastics into smaller fragments, commonly known as microplastics. These microplastics are subsequently transported into marine environments through runoff (<xref ref-type="bibr" rid="B44">Nguyen et&#xa0;al., 2025</xref>). In fact, the plastic material size ranging from 1 &#x3bc;M to 5&#xa0;mm has been classified as microplastic (<xref ref-type="bibr" rid="B12">Cole et&#xa0;al., 2011</xref>). Microplastics are divided into primary and secondary microplastics (<xref ref-type="bibr" rid="B6">Bhuyan et al., 2021</xref>). The main source of primary plastic involves cosmetic products, personal healthcare products, and children&#x2019;s products (<xref ref-type="bibr" rid="B26">Hartmann et al., 2019</xref>). However, the major sources of secondary plastics are fragmented products produced via the physical fragmentation as well as the biological and chemical degradation of large-sized plastic material (<xref ref-type="bibr" rid="B58">Sait et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B75">Yuan et&#xa0;al., 2022</xref>). It is important to mention that most current studies on microplastic toxicity are increasingly focused on elucidating the underlying mechanisms responsible for their toxicity (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>). The latter area of microplastics research is important considering that some of the chemicals associated with plastic contamination are able to disrupt the endocrine system in vertebrates, including fish and mammals (<xref ref-type="bibr" rid="B23">Gugliandolo et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B19">Folbert et&#xa0;al., 2022</xref>; <xref ref-type="bibr" rid="B13">Corti et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B62">Sharma et al., 2024</xref>). Furthermore, microplastics are readily assimilated by plankton, which can serve as a transfer route to secondary and tertiary consumers in the marine food chain, potentially leading to consequences for humans, the final consumers (<xref ref-type="bibr" rid="B24">Gunaalan et&#xa0;al., 2023</xref>). It should be noted that the smaller the size of microplastics, the more the toxicological consequences (<xref ref-type="bibr" rid="B38">Markic et al., 2020</xref>; <xref ref-type="bibr" rid="B65">Tang, 2024</xref>).</p>
<p>On the other hand, <italic>Posidonia oceanica</italic> (PO) is known as marine seagrass and an endemic species to Mediterranean Sea (<xref ref-type="bibr" rid="B17">De Luca et&#xa0;al., 2025</xref>). The shoots of <italic>P. oceanica</italic> constitute structurally complex ecosystems, providing adequate living conditions and ecological niches for a significant number of organisms (<xref ref-type="bibr" rid="B9">Boudouresque et al., 2016</xref>). On the beaches of the Mediterranean Sea, one often finds ball-shaped clumps of plant debris. These natural formations are called &#x201c;aegagropiles (EG),&#x201d; and they are usually made from fibers of the seagrass <italic>Posidonia oceanica</italic>, whose size diverges from millimeters to centimeters (<xref ref-type="bibr" rid="B70">Verhille and Le Gal, 2018</xref>). These balls occupied large areas, notably after storms. Research revealed that EG are formed by hydrodynamic flows and composed of different plant fibrous elements (PO) and sand grains (<xref ref-type="bibr" rid="B39">Matheson et al., 2017</xref>). The leaf cells of <italic>Posidonia</italic> are distinguished by their thin and lignified walls, and thus the fibers offer the rigidity necessary to form EG (<xref ref-type="bibr" rid="B50">Pi&#xf1;eiro-Juncal et&#xa0;al., 2021</xref>).</p>
<p>PO plays an important ecological role by acting as a sink for contaminants, particularly by storing them in its roots and shoots, thereby helping in reducing their availability in the marine environment (<xref ref-type="bibr" rid="B64">Tahir et al., 2019</xref>; <xref ref-type="bibr" rid="B61">Sghaier et&#xa0;al., 2025</xref>). It actively removes certain pollutants, sequestering a portion within its tissues and thereby limiting their transfer through trophic networks. Moreover, several studies highlight the role of <italic>P. oceanica</italic> as an effective bioindicator of marine pollution, as well as a long-term archive of contaminants through accumulation in the matte (the dense, long-lasting underground structure formed by intertwined rhizomes, roots, and trapped sediments), which serves as an environmental memory of pollutant inputs (<xref ref-type="bibr" rid="B66">Telesca et al., 2015</xref>; <xref ref-type="bibr" rid="B61">Sghaier et&#xa0;al., 2025</xref>).</p>
<p>In this context, it is essential to identify effective strategies to mitigate the impact of chemical pollution on seagrass beds. Although various studies have investigated the distribution of microplastics in specific geographical regions and others have examined their effects on certain marine organisms, global knowledge remains limited due to the relatively small number of in-depth studies focusing on natural bioaggregates such as aegagropiles (EGs) and their role in trapping and transferring microplastics. Therefore, the present study aims to assess the accumulation of microplastics in aegagropiles and to identify the most abundant polymers present in the marine environment.</p>
</sec>
<sec id="s2" sec-type="materials|methods">
<title>Material and methods</title>
<sec id="s2_1">
<title>Sampling</title>
<p><italic>P. oceanica</italic> aegagropiles were sampled at four different sites, four from the North of Tunisia including Bizerte (latitude 37&#xb0;17&#x2032;45.02&#x2033;N, longitude 9&#xb0;52&#x2032;23.22&#x2033;E) (S1), and Hammamet (longitude 10&#xb0;32&#x2032;29.84&#x2033;E, latitude 36&#xb0;22&#x2032;13.76&#x2033;N) (S2), and from the east-central Tunisia at two different sites: Mahdia (latitude 35&#xb0;30&#x2032;56.39&#x2033;N, longitude 11&#xb0; 2&#x2032;56.66&#x2033;E) (S3) and Chebba (latitude 35&#xb0;26&#x2032;56.96&#x2033;N, longitude 11&#xb0; 0&#x2032;16.78&#x2033;E) (S4) (<xref ref-type="fig" rid="f1"><bold>Figure&#xa0;1</bold></xref>). These sites are characterized by tourism and fishing activities. Aegagropiles were manually collected in triplicate from seagrass banquettes (dense accumulations of dead seagrass leaves along the shoreline), as illustrated in <xref ref-type="fig" rid="f2"><bold>Figure&#xa0;2</bold></xref>. The samples were placed in sterile sample bags and transported to the Laboratory of Marine Ecotoxicology at National Institute of Marine sciences and Technology for subsequent laboratory analysis.</p>
<fig id="f1" position="float">
<label>Figure&#xa0;1</label>
<caption>
<p>Map showing the four sampling areas sites along the coast of Tunisia. (S1) Bizerte, (S2) Hammamet, (S3) Mahdia, (S4) Chebba.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fmars-12-1663783-g001.tif">
<alt-text content-type="machine-generated">Map of Tunisia showing geographic locations with markers S1 to S4. Cities like Tunis and Sousse are labeled. A yellow border outlines the country, and a scale indicates 500 kilometers. A north arrow is present.</alt-text>
</graphic>
</fig>
<fig id="f2" position="float">
<label>Figure&#xa0;2</label>
<caption>
<p>Aegagropiles collected from Tunisian beaches: <bold>(A)</bold> Bizerte, <bold>(B)</bold> Mahdia, <bold>(C)</bold> close-up image of an aegagropile.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fmars-12-1663783-g002.tif">
<alt-text content-type="machine-generated">Panel A shows a sandy beach with scattered seaweed and fibrous balls. Panel B depicts a strandline with numerous small fibrous balls mixed with debris. Panel C displays three close-up fibrous balls on a flat surface.</alt-text>
</graphic>
</fig>
</sec>
<sec id="s2_2">
<title>Microplastic extraction</title>
<p>In the laboratory, the samples underwent a thorough cleaning process with distilled water to remove any sediment, followed by air drying at room temperature (25&#xa0;&#xb0;C) and low humidity for several days. The aegagropiles of <italic>P. oceanica</italic> were weighed using an analytical balance with a sensitivity of 0.01 mg. To dissolve and digest the organic matter, two methods were applied following <xref ref-type="bibr" rid="B59">Sanchez-Vidal et&#xa0;al. (2021)</xref>, with some modifications. In the first method, a solution containing 10% HCl and 30 mL of 30% H<sub>2</sub>O<sub>2</sub> was introduced. After a reaction period of 48&#xa0;h, a KOH solution (10%) was added to promote chemical digestion. The samples were dried in the oven at 50&#xa0;&#xb0;C for 1 week. Then, the resulting mixture containing solid residues and MP was sieved. The contents of the sieves were transferred to a glass jar for decanting. The supernatant was filtered under vacuum through a Whatman glass fiber filter with a diameter of 47&#xa0;mm (GF/D, with a particle retention size of 2.7 &#xb5;m). The filter cake was completely rinsed with deionized water and then dried in an oven at 60&#xb0;C, and the membrane was stored in a glass Petri dish for air drying at room temperature.</p>
<p>With the second method, the EGs were carefully disentangled, and the fibers were sieved at 5, 0.36, and 0.2&#xa0;mm using a stainless-steel sieve. The contents retained on the sieves were treated with 4&#x2013;6 mL of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>, 30%), followed by 10% hydrochloric acid (HCl) to remove most of the organic matter and calcium carbonate. The samples were then dried in an oven at 50&#xb0;C for more than 24&#xa0;h.</p>
</sec>
<sec id="s2_3">
<title>Quality control</title>
<p>During the sampling, no plastic tools and materials were used. Additionally, the laboratory contamination was assessed by placing a moist filter over an opened Petri dish. The operators were required to wear cotton coats to further reduce the risk of contamination. Prior to use, the filters were meticulously inspected under a microscope to ensure they were free from any airborne microplastic particles. When handling samples, stainless-steel forceps were used to maintain the integrity of the samples.</p>
</sec>
<sec id="s2_4">
<title>Microplastics identifications</title>
<p>All extracted plastic particles were picked out with metal tweezers into a 90-mm Petri dish containing a black and white background that enabled high contrast with plastic colors and types, which was photographed. The Petri dishes were inspected for plastic debris under a Leica M60 stereomicroscope (Leica Microsystems AG, Glattbrugg, Switzerland) equipped with a CMOS microscope camera and a 6:1 zoom system, offering a continuous magnification range from 2&#xd7; to 5&#xd7;, with engageable click stops for precise settings (see <xref ref-type="bibr" rid="B27">Hassen et&#xa0;al., 2023</xref>).</p>
</sec>
<sec id="s2_5">
<title><sup>1</sup>H-NMR characterization</title>
<p>Using proton nuclear magnetic resonance spectroscopy (<sup>1</sup>H-NMR) characterization, MP particles were dissolved in a suitable deuterated solvent. Deuterated dimethyl sulfoxide (DMSO-d6) (99.8 atom %D) and deuterated chloroform (CDCl3) (99.8 atom %D, stab. with Ag) from Deutero, Germany, were used.</p>
<p>The NMR measurements were performed using a Bruker Avance III 300 MHz spectrometer. Data were recorded at room temperature, at a spinning rate of 14 kHz and with a pulse length of 90&#xb0; and 3.25 &#x3bc;s with a 5-s interval between scans (see <xref ref-type="bibr" rid="B47">Peez and Imhof, 2020</xref>).</p>
</sec>
<sec id="s2_6">
<title>FT-IR characterization</title>
<p>The Fourier-transform infrared (FTIR) measurements were carried out on a Perkin Elmer Spectrum BX FTIR device (Perkin Elmer, USA), utilizing a Golden Gate single reflection diamond ATR system (SpecacLda, USA). All spectra were obtained using 64 scans each and a resolution of 4 cm<sup>&#x2212;1</sup>, within the 4,000&#x2013;450-cm<sup>&#x2212;1</sup> range (see <xref ref-type="bibr" rid="B7">Blindheim and Ruwoldt, 2023</xref>).</p>
</sec>
</sec>
<sec id="s3" sec-type="results">
<title>Results</title>
<sec id="s3_1">
<title>Microscopy identification</title>
<p>An examination of the colors and types of microplastics trapped in the EGs (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3</bold></xref>) revealed that the plastic items mainly consisted of fragments and threads. These were of various sizes and colors and were found to be intertwined within the examined EGs.</p>
<fig id="f3" position="float">
<label>Figure&#xa0;3</label>
<caption>
<p>Microplastic particles found in EG extract samples under a stereomicroscope. <bold>(A)</bold> Bizerte, <bold>(B)</bold> Hammamet, <bold>(C)</bold> Chebba, <bold>(D)</bold> Mahdia.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fmars-12-1663783-g003.tif">
<alt-text content-type="machine-generated">Microscopic images labeled A to D, showing elongated fibrous structures under magnification. Black arrows indicate specific fibers. Each sub-image includes a scale bar measuring one hundred micrometers for reference.</alt-text>
</graphic>
</fig>
<p>The frequent colors of microplastics were mainly blue and red. In site S1, the found microplastic elements included blue, black, and transparent fragments and filaments with red and yellow colors (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3A</bold></xref>).</p>
<p>At the S2 site (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3B</bold></xref>), red, blue, and transparent filaments were detected with the presence of black and blue fragments. Concerning site S3 (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3C</bold></xref>), black and yellow fragments are essentially observed with the presence of transparent filaments. For site S4, black and red fragments are identified (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3D</bold></xref>).</p>
</sec>
<sec id="s3_2">
<title>FTIR results</title>
<p>According to the literature (<xref ref-type="table" rid="T1"><bold>Table&#xa0;1</bold></xref>), the results obtained by FTIR showed intense absorptions at 3,250 (hydrogen bonded N&#x2013;H stretch), 2,917-2,840 cm<sup>&#x2212;1</sup> (ns CH<sub>2</sub>), 1,722-1,529 (amide I), 1,354, 1,249 (amide III), 1,152-1,003 (C&#x2013;C str), 800-700 (CH<sub>2</sub> rocking), and 694-500 (Aromatic CH out-of-plane bending) cm<sup>&#x2212;1</sup>.</p>
<table-wrap id="T1" position="float">
<label>Table&#xa0;1</label>
<caption>
<p>FTIR characteristic peak assignments in cm<sup>-1</sup>for various types of MPs.</p>
</caption>
<table frame="hsides">
<thead>
<tr>
<th valign="middle" align="left">POLYMER</th>
<th valign="middle" align="left">Characteristic peaks (cm<sup>-1</sup> )</th>
<th valign="middle" align="left">Assignement</th>
<th valign="middle" align="left">References</th>
</tr>
</thead>
<tbody>
<tr>
<td valign="middle" rowspan="7" align="left">Low density polyethylene (LDPE)</td>
<td valign="middle" align="left">2915</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
<td valign="middle" rowspan="7" align="left"><xref ref-type="bibr" rid="B42">Nishikida and Coates, 2003</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B3">Asensio et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B40">Mecozzi et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">2845</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1467</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1462</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1377</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">730</td>
<td valign="middle" align="left">CH2 rocking</td>
</tr>
<tr>
<td valign="middle" align="left">717</td>
<td valign="middle" align="left">CH2 rocking</td>
</tr>
<tr>
<td valign="middle" rowspan="4" align="left">Polyethylene terephthalate (PET)</td>
<td valign="middle" align="left">1713</td>
<td valign="middle" align="left">C=O stretching</td>
<td valign="middle" rowspan="4" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B3">Asensio et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B40">Mecozzi et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">1241</td>
<td valign="middle" align="left">C&#x2013;O stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1094</td>
<td valign="middle" align="left">C&#x2013;O stretching Aromatic</td>
</tr>
<tr>
<td valign="middle" align="left">720</td>
<td valign="middle" align="left">CH out-of plane bending</td>
</tr>
<tr>
<td valign="middle" rowspan="10" align="left">Polypropylene (PP)</td>
<td valign="middle" align="left">2950</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
<td valign="middle" rowspan="10" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B3">Asensio et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B40">Mecozzi et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">2838</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
</tr>
<tr>
<td valign="middle" align="left">2915</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1455</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1377</td>
<td valign="middle" align="left">CH3 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1166</td>
<td valign="middle" align="left">CH bending, CH3 rocking, C&#x2013;C stretching</td>
</tr>
<tr>
<td valign="middle" align="left">997</td>
<td valign="middle" align="left">CH3 rocking, CH3 bending, CH bending</td>
</tr>
<tr>
<td valign="middle" align="left">972</td>
<td valign="middle" align="left">CH3 rocking, C&#x2013;C stretching</td>
</tr>
<tr>
<td valign="middle" align="left">840</td>
<td valign="middle" align="left">CH2 rocking, C&#x2013;CH3 stretching</td>
</tr>
<tr>
<td valign="middle" align="left">808</td>
<td valign="middle" align="left">CH2 rocking, C&#x2013;C stretching, C&#x2013;CH stretching Aromatic</td>
</tr>
<tr>
<td valign="middle" rowspan="8" align="left">Polystyrene (PS)</td>
<td valign="middle" align="left">3024</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
<td valign="middle" rowspan="7" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B3">Asensio et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B40">Mecozzi et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">2847</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1601</td>
<td valign="middle" align="left">Aromatic ring stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1492</td>
<td valign="middle" align="left">Aromatic ring stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1451</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1027</td>
<td valign="middle" align="left">Aromatic CH bending</td>
</tr>
<tr>
<td valign="middle" align="left">694</td>
<td valign="middle" align="left">Aromatic CH out-of plane bending</td>
</tr>
<tr>
<td valign="middle" align="left">537</td>
<td valign="middle" align="left">Aromatic ring out-of plane bending</td>
<td valign="middle" rowspan="7" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" rowspan="6" align="left">Polyvinyl chloride (PVC)</td>
<td valign="middle" align="left">1427</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1331</td>
<td valign="middle" align="left">CH bending</td>
</tr>
<tr>
<td valign="middle" align="left">1255</td>
<td valign="middle" align="left">CH bending</td>
</tr>
<tr>
<td valign="middle" align="left">1099</td>
<td valign="middle" align="left">C&#x2013;C stretching</td>
</tr>
<tr>
<td valign="middle" align="left">966</td>
<td valign="middle" align="left">CH2 rocking</td>
</tr>
<tr>
<td valign="middle" align="left">616</td>
<td valign="middle" align="left">C&#x2013;Cl stretching</td>
</tr>
<tr>
<td valign="middle" rowspan="7" align="left">Ethylene vinyl acetate (EVA)</td>
<td valign="middle" align="left">2917</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
<td valign="middle" rowspan="7" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B3">Asensio et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">2848</td>
<td valign="middle" align="left">C&#x2013;H stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1740</td>
<td valign="middle" align="left">C = O stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1469</td>
<td valign="middle" align="left">CH2 bending, CH3 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1241</td>
<td valign="middle" align="left">C (=O) O stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1020</td>
<td valign="middle" align="left">C&#x2013;O stretching</td>
</tr>
<tr>
<td valign="middle" align="left">720</td>
<td valign="middle" align="left">CH2 rocking</td>
</tr>
<tr>
<td valign="middle" rowspan="10" align="left">Nylon (all polyamides)</td>
<td valign="middle" align="left">3298</td>
<td valign="middle" align="left">N&#x2013;H stretching</td>
<td valign="middle" rowspan="10" align="left"><xref ref-type="bibr" rid="B72">Verleye et&#xa0;al., 2001</xref>; <xref ref-type="bibr" rid="B43">Noda et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B40">Mecozzi et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B33">Jung et&#xa0;al., 2018</xref></td>
</tr>
<tr>
<td valign="middle" align="left">2932</td>
<td valign="middle" align="left">CH stretching</td>
</tr>
<tr>
<td valign="middle" align="left">2858</td>
<td valign="middle" align="left">CH stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1634</td>
<td valign="middle" align="left">C =O stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1538</td>
<td valign="middle" align="left">NH bending, C&#x2013;N stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1464</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1372</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">1274</td>
<td valign="middle" align="left">NH bending, C&#x2013;N stretching</td>
</tr>
<tr>
<td valign="middle" align="left">1199</td>
<td valign="middle" align="left">CH2 bending</td>
</tr>
<tr>
<td valign="middle" align="left">687</td>
<td valign="middle" align="left">NH bending, C=O bending</td>
</tr>
</tbody>
</table>
</table-wrap>
<p><xref ref-type="fig" rid="f4"><bold>Figure&#xa0;4</bold></xref> shows different types of polymers determined in each site. In site S1 (<xref ref-type="fig" rid="f4"><bold>Figure&#xa0;4A</bold></xref>), bands at 3,300, 2,992, 1,700, 1,500, 1,250, 1,050, 750, and 550 cm<sup>&#x2212;1</sup> are related to polysterene (PS), polyethylene terephthalate (PET), and ethylene-vinyl acetate (EVA). Similarly, in site S2 (<xref ref-type="fig" rid="f4"><bold>Figure&#xa0;4B</bold></xref>), the collected spectra revealed that bands at 2,875, 1,750, 1,650, 1,500, 1,250, 1,000, 750, 550, and 500 cm<sup>&#x2212;1</sup> demonstrated the existence of polyvinylchloride (PVC), propylene (PP), and PET.</p>
<fig id="f4" position="float">
<label>Figure&#xa0;4</label>
<caption>
<p>Polymers of microplastic obtained from FTIR spectroscopy reads found in <bold>(A)</bold> Bizerte, <bold>(B)</bold> Hammamet, <bold>(C)</bold> Chebba, and <bold>(D)</bold> Mahdia (assigned characteristic peaks in cm<sup>&#x2212;1</sup>).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fmars-12-1663783-g004.tif">
<alt-text content-type="machine-generated">Four infrared spectroscopy graphs labeled A, B, C, and D demonstrate transmittance versus wavenumber for regions Bizerte, Hammamet, Mahdia, and Chebba. Each graph displays differing transmittance patterns across the wavenumber range of 0 to 4500 cm^-1.</alt-text>
</graphic>
</fig>
<p>At site S3 in the Tunisian east-center, as depicted in <xref ref-type="fig" rid="f4"><bold>Figure&#xa0;4C</bold></xref>, bands observed at 2,900, 1,800, 1,650, 1,300, 1,150, and 1,000 cm<sup>&#x2212;1</sup> indicated the presence of PS and EVA. However, in site S4 (<xref ref-type="fig" rid="f4"><bold>Figure&#xa0;4D</bold></xref>), the spectrum revealed the existence of PVC and PS, characterized by bands at 2,900, 1,800, 1,750, 1,600, 1,400, 1,100, 950, and 600 cm<sup>&#x2212;1</sup>.</p>
</sec>
<sec id="s3_3">
<title><sup>1</sup>H-NMR spectroscopy results</title>
<p>The spectra obtained for each compound separately were determined according to previous studies related to microplastic identifications (<xref ref-type="table" rid="T2"><bold>Table&#xa0;2</bold></xref>). According to the literature, in site S1, three types of microplastics are determined, namely, PS, PA, and PVC (<xref ref-type="fig" rid="f5"><bold>Figure&#xa0;5A</bold></xref>). In fact, two signals can be clearly assigned to PVC, one of which with a chemical shift of 4.6-4.2 ppm (H1) corresponds to the <italic>&#x3b1;</italic>-Cl H atoms and the other in the range of 2.5-2.1 ppm (H2) matches with the <italic>&#x3b2;</italic>-Cl H atom. With respect to PS, four signals were assigned: 1.87 ppm (H1) 1.46 ppm (H2), 6.4-6.8 ppm (H3), and 7.11 ppm (H4, 5). It is worthwhile to note that PA signals at high field with chemical shifts between 3.2 and 1.1 ppm can be assigned to protons H1-H5. The signal with the chemical shift of 3.2-3.0 ppm (H1) aligns with the <italic>&#x3b1;</italic>-NH H atoms and the signal in the range of 2.3-2.15 ppm (H2) matches the protons H2 of the <italic>&#x3b1;</italic>-CO group. Signals in the range of 1.65-1.1 ppm represent CH2 groups H3-H5 of the polymer chain.</p>
<table-wrap id="T2" position="float">
<label>Table&#xa0;2</label>
<caption>
<p><sup>1</sup>H- NMR characteristic signal assignments in ppm for various types of MPs.</p>
</caption>
<table frame="hsides">
<thead>
<tr>
<th valign="middle" align="center">Polymer</th>
<th valign="middle" align="center">ppm</th>
<th valign="middle" align="center">Assignement</th>
<th valign="middle" align="center">References</th>
</tr>
</thead>
<tbody>
<tr>
<td valign="middle" rowspan="3" align="center">PET</td>
<td valign="middle" align="center">8.119</td>
<td valign="middle" align="center">H1</td>
<td valign="middle" rowspan="2" align="center"><xref ref-type="bibr" rid="B45">Papini et&#xa0;al., 2022</xref></td>
</tr>
<tr>
<td valign="middle" align="center">4.713</td>
<td valign="middle" align="center">H2</td>
</tr>
<tr>
<td valign="middle" align="center">1.33</td>
<td valign="middle" align="center">H2</td>
<td valign="middle" rowspan="2" align="center"><xref ref-type="bibr" rid="B48">Peez et&#xa0;al., 2019</xref></td>
</tr>
<tr>
<td valign="middle" align="center">PE</td>
<td valign="middle" align="center">0.93</td>
<td valign="middle" align="center">H1</td>
</tr>
<tr>
<td valign="middle" rowspan="5" align="center">PS</td>
<td valign="middle" align="center">1.878</td>
<td valign="middle" align="center">H1</td>
<td valign="middle" rowspan="4" align="center"><xref ref-type="bibr" rid="B45">Papini et&#xa0;al., 2022</xref></td>
</tr>
<tr>
<td valign="middle" align="center">1.463</td>
<td valign="middle" align="center">H2</td>
</tr>
<tr>
<td valign="middle" align="center">6.400-6.800</td>
<td valign="middle" align="center">H3</td>
</tr>
<tr>
<td valign="middle" align="center">7.112</td>
<td valign="middle" align="center">H4,5</td>
</tr>
<tr>
<td valign="middle" align="center">3.2-3.0</td>
<td valign="middle" align="center">H1</td>
<td valign="middle" rowspan="4" align="center"><xref ref-type="bibr" rid="B47">Peez and Imhof, 2020</xref></td>
</tr>
<tr>
<td valign="middle" rowspan="4" align="center">PA</td>
<td valign="middle" align="center">2.3-2.15</td>
<td valign="middle" align="center">H2</td>
</tr>
<tr>
<td valign="middle" align="center">1.65-1.1</td>
<td valign="middle" align="center">H3-H5</td>
</tr>
<tr>
<td valign="middle" align="center">7.31</td>
<td valign="middle" align="center">(amines)</td>
</tr>
<tr>
<td valign="middle" align="center">0.74</td>
<td valign="middle" align="center">H1</td>
<td valign="middle" rowspan="3" align="center"><xref ref-type="bibr" rid="B29">Hero and Kali, 2020</xref></td>
</tr>
<tr>
<td valign="middle" rowspan="2" align="center">PP</td>
<td valign="middle" align="center">1.19</td>
<td valign="middle" align="center">H2</td>
</tr>
<tr>
<td valign="middle" align="center">1.48</td>
<td valign="middle" align="center">H3</td>
</tr>
<tr>
<td valign="middle" rowspan="4" align="center">PVC</td>
<td valign="middle" align="center">2.000-2.500</td>
<td valign="middle" align="center">H2</td>
<td valign="middle" rowspan="2" align="center"><xref ref-type="bibr" rid="B45">Papini et&#xa0;al., 2022</xref></td>
</tr>
<tr>
<td valign="middle" align="center">4.250-4.670</td>
<td valign="middle" align="center">H1</td>
</tr>
<tr>
<td valign="middle" align="center">0.86</td>
<td valign="middle" align="center">H1</td>
<td valign="middle" rowspan="5" align="center"><xref ref-type="bibr" rid="B55">Ren et&#xa0;al., 2019</xref></td>
</tr>
<tr>
<td valign="middle" align="center">1.25</td>
<td valign="middle" align="center">H2</td>
</tr>
<tr>
<td valign="middle" rowspan="3" align="center">EVA</td>
<td valign="middle" align="center">1.74</td>
<td valign="middle" align="center">H3</td>
</tr>
<tr>
<td valign="middle" align="center">2.01</td>
<td valign="middle" align="center">H4</td>
</tr>
<tr>
<td valign="middle" align="center">4.85</td>
<td valign="middle" align="center">H5</td>
</tr>
</tbody>
</table>
</table-wrap>
<fig id="f5" position="float">
<label>Figure&#xa0;5</label>
<caption>
<p><sup>1</sup>H-NMR spectra of plastic polymers found in the Tunisian coastlines of the four different sites <bold>(A)</bold> Bizerte, <bold>(B)</bold> Hammamet, <bold>(C)</bold> Chebba, and <bold>(D)</bold> Mahdia (assigned chemical shifts in ppm).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fmars-12-1663783-g005.tif">
<alt-text content-type="machine-generated">Spectral data displaying four labeled panels: A, B, C, and D. Each panel includes line graphs with peaks and troughs, representing chemical shifts in parts per million (ppm). Annotations provide detailed information on sample parameters.</alt-text>
</graphic>
</fig>
<p>In site S2 (<xref ref-type="fig" rid="f5"><bold>Figure&#xa0;5B</bold></xref>), EVA, PP, and PE can be observed. As regards EVA, five signals were assigned at 0.86 to H1, 1.25 (H2), 1.74 (H3), 2.01 (H4), and 4.85 to (H5). The PE can be assigned to two signals. The signal at 1.33 ppm (H2) corresponds to the protons of the CH<sub>2</sub> groups and the one at 0.93 ppm (H1) corresponds to the protons of the CH<sub>3</sub> group. Three signals defined PP and can be seen in the NMR spectrum at 0.74, 1.19, and 1.48 ppm, belonging to the methyl, methylene, and methine groups of oligo/polymeric propylene.</p>
<p>While PVC, EVA, and PP were determined in site S3 (<xref ref-type="fig" rid="f5"><bold>Figure&#xa0;5C</bold></xref>), PA and PP were detected in site S4 (<xref ref-type="fig" rid="f5"><bold>Figure&#xa0;5D</bold></xref>).</p>
</sec>
</sec>
<sec id="s4" sec-type="discussion">
<title>Discussion</title>
<p>Microplastics (MPs) are composed of various polymers, and their distribution in the environment depends on both morphological and chemical characteristics (<xref ref-type="bibr" rid="B21">Giaganini et al., 2023</xref>). Understanding these characteristics is important because the shape and size of MPs can influence their transport and accumulation in different environments. For instance, morphological traits may help infer how MPs are distributed across coastal areas (horizontally), whereas polymer type can affect their behavior in the water column (vertically). Although our study does not directly assess horizontal or vertical distribution, we adopted a similar approach by characterizing MPs in eagagropiles (EGs) based on morphology (e.g., fragments, threads) and polymer composition, using microscopy and spectroscopy. This characterization helps to understand the types and potential sources of MPs accumulating along the Tunisian coasts. Previous studies have similarly emphasized the importance of such classification (e.g., <xref ref-type="bibr" rid="B2">Anderson et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B68">Veerasingam et&#xa0;al., 2020</xref>). Microscopy identification of microplastics (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3</bold></xref>) showed that fibers particularly red (100%), transparent (100%), and blue (75%) were the most commonly observed in all samples. While the black fragment was revealed in two sites, the yellow fragment was identified in one site (<xref ref-type="fig" rid="f3"><bold>Figure&#xa0;3</bold></xref>). However, <xref ref-type="bibr" rid="B5">Ben Ismail et&#xa0;al. (2022)</xref> reported different MP forms in water samples collected in the Gulf of Gabes, with fragments being the most abundant plastic form, whereas fibers, pellets, films, and foams were detected in only a small fraction. This observation was confirmed by <xref ref-type="bibr" rid="B76">Zayen et&#xa0;al. (2020)</xref>, and five different categories of MP forms, fragments, films, filaments, pellets, and foam, were found in the water of the same site. The color of MP items was white in the majority, whereas blue, black, green, and red items accounted on average for 9%, 5%, 5%, and 3% of the total MPs, respectively.</p>
<p>In other studies, other identified colors, including yellow, orange, gray, and purple, were also present within MPs (4%; 344 of total MPs) (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>). <xref ref-type="bibr" rid="B15">Dahl et&#xa0;al. (2021)</xref> examined soil collected in PO meadows at three locations along the Spanish Mediterranean coast and reported that the most common particle colors were transparent (30%), white (18%), and green (15%), and the dominant forms were irregular (58%) and flat (30%). In addition, according to several studies, transparent particles of MPs represented approximately 20% to 70% of the total plastic-like particles, initially revealed by stereomicroscopy and afterward classified by other methods (<xref ref-type="bibr" rid="B28">He et&#xa0;al., 2025</xref>; <xref ref-type="bibr" rid="B63">Song et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B37">Mariano et&#xa0;al., 2021</xref>). Likewise, it is difficult to distinguish between natural and synthetic fibers when using a stereomicroscope, which are prevalent constituents in water, soil, and biota (<xref ref-type="bibr" rid="B36">Lusher et&#xa0;al., 2013</xref>; <xref ref-type="bibr" rid="B16">de los Santos et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B37">Mariano et&#xa0;al., 2021</xref>). This allows for the rapid identification of the shape, size, and color of the particles before subsequent characterization by other techniques. The use of microscopy combined with additional methods, such as spectroscopy, enhances the accuracy and comprehensiveness of microplastic analysis (<xref ref-type="bibr" rid="B37">Mariano et&#xa0;al., 2021</xref>).</p>
<p>Within the same vein, to determine the type of microplastics accumulated in EGs, FTIR and NMR analyses were carried out (<xref ref-type="fig" rid="f4"><bold>Figures&#xa0;4</bold></xref>, <xref ref-type="fig" rid="f5"><bold>5</bold></xref>). Using two spectroscopy methods allowed for defining the polymers identified in the samples. Different types of microplastics are identified, principally polysterene, propylene, ethyl-vinyl acetate, polyamide, and polyvinyl chloride which are mainly the most found.</p>
<p>The dominance of polymers such as polysterene (PS), polyethylene terephthalate (PET), polyvinyl chloride (PVC), and ethyl-vinyl acetate (EVA) reflects their widespread use in everyday items ranging from packaging, textiles, to construction materials commonly found in the region (<xref ref-type="bibr" rid="B25">Gurjar et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B31">Jamsek et&#xa0;al., 2024</xref>). Coastal tourism, which plays a vital role in the local economy, often brings seasonal surges of plastic waste, whereas urban runoff, wastewater discharge, and river inputs transport land-based sources into the marine ecosystem (<xref ref-type="bibr" rid="B25">Gurjar et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B31">Jamsek et&#xa0;al., 2024</xref>).</p>
<p>Several previous studies have confirmed similar polymer distributions in the Mediterranean and other regions (<xref ref-type="bibr" rid="B54">Pourebrahimi and Pirooz, 2023</xref>). According to <xref ref-type="bibr" rid="B76">Zayen et&#xa0;al. (2020)</xref>, the common MP items were attributed to polyolefins, basically polyethylene and reformulated polyethylene, as well as polypropylene (PP) and ethylene&#x2013;propylene copolymers, from water samples collected in near-surface waters of the Gulf of Gabes. Based on water samples taken from the Gulf of Gabes, <xref ref-type="bibr" rid="B5">Ben Ismail et&#xa0;al. (2022)</xref> characterized 11 different polymer typologies determined as PE, constituting the majority of MPs, followed by PP. Less frequent polymers included (&lt;6%) PS, polyvinyl alcohol (PVA), polyamides (PA), acrylic (Acr), ethylene-vinyl acetate (EVA), polyvinyl chloride (PVC), ethylene propylene diene monomer (EPDM), and polyesters (mainly PET). In addition, <xref ref-type="bibr" rid="B5">Ben Ismail et&#xa0;al. (2022)</xref> admitted that from biota samples, a mixture of PE and EVA was determined. Nevertheless, <xref ref-type="bibr" rid="B49">Pietrelli et&#xa0;al. (2017)</xref> reported that PE and PP were found in significantly higher amounts in sand, whereas PE, nylon, polyester, and microfibers (as pills) were more frequently detected in EGs from samples collected along the central coast of Italy.</p>
<p>In this context, in Tarragona coastal regions, PP and PE fragments were the prevalent MPs on beaches, whereas polyester fibers were dominated in the bottom sediments and saltwater (<xref ref-type="bibr" rid="B18">Exp&#xf3;sito et&#xa0;al., 2021</xref>). The abundance of fiber balls coupled with bottom sediments, organic materials, and plankton hid the true fibers present in each reservoir (<xref ref-type="bibr" rid="B18">Exp&#xf3;sito et&#xa0;al., 2021</xref>). Nevertheless, other plastic polymers have been detected from marine sediment from the German coast showing the presence of PP in each sample (<xref ref-type="bibr" rid="B35">Lorenz, 2014</xref>). PS was also present throughout the samples, although it occurred less frequently than other polymer types. Denser polymers, such as PVC and PVA, have been detected alongside low-density polymers like PE. In other studies, for example, in the Venice lagoon and in PO meadows adjacent to agricultural hinterland in Spain, PE and PP were revealed as the most abundant materials in seawater and sediments. Additionally, fragments and filaments were the most common forms determined, as signaled by <xref ref-type="bibr" rid="B22">Grego et&#xa0;al. (2022)</xref>.</p>
<p>Generally, PP, PE, and PET were admitted as common polymer types in the marine environment (<xref ref-type="bibr" rid="B78">Zhou et&#xa0;al., 2021</xref>). More deeply, PE and PP are frequently detected materials due to their low density (<xref ref-type="bibr" rid="B46">Pedrotti et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B74">Yao et&#xa0;al., 2024</xref>), which permits their floating and immersion. These polymers are widely used as polymers in various commodities like packaging, household plastic waste, and numerous personal care and cosmetic products (<xref ref-type="bibr" rid="B12">Cole et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B52">Plastic Europe, 2020</xref>).</p>
<p>Beyond the Mediterranean, studies such as <xref ref-type="bibr" rid="B4">Ballent et&#xa0;al. (2016)</xref> reported the presence of various polymers including PE, PS, PU, PVC, and PSS in high abundance in Canadian Lake Ontario, with other polymers like PET, nylon, and ABS found in smaller quantities. Although our study focuses on the Tunisian coast, these findings support the global ubiquity and diversity of microplastic polymer types, aligning with the polymeric variety we observed in the EGs.</p>
<p>The identified microplastics differ from one site to another, depending on the specific activities in these regions, such as tourism, agriculture, fishing, and industry. In fact, it is difficult to determine their origin because a single polymer may come from several products and have a huge range of sources (<xref ref-type="bibr" rid="B4">Ballent et&#xa0;al., 2016</xref>). PE, PP, PS, PA, PVC, EVA, and nylon are the most dominating forms of microplastic distributed in marine, freshwater, and estuarine environments (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>). PE and its derivatives are generally used in fishing gears (<xref ref-type="bibr" rid="B73">Wang et&#xa0;al., 2021</xref>), food packaging, agricultural film, and plastic bottles and plastic bags (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>). However, PP is used in plastic containers, food packaging, carpets, and pipes (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>). While PS is used in food containers, rubber tires, and boat hulls, surfboards, bathtubs, and shower enclosures, EVA is used as padding in the equipment of various sports such as ski boots, bicycle saddles, waterski boots, fishing rods, and fishing-reel handles (<xref ref-type="bibr" rid="B77">Zhang et&#xa0;al., 2022</xref>).</p>
<p>Anthropogenic pressures, combined with insufficient waste management and densely populated coastlines typical of the Mediterranean basin, contribute significantly to the observed microplastic contamination patterns in <italic>Posidonia oceanica</italic> seagrass aegagropiles (EGs). Therefore, seagrass meadows constitute an area of microplastic accumulation, which accords well with the findings obtained by <xref ref-type="bibr" rid="B14">Cozzolino et&#xa0;al. (2020)</xref>, who reported higher macroplastic presence in vegetated coastal areas compared with unvegetated ones. Such variation in microplastic distribution within vegetated zones is influenced by multiple factors, including anthropogenic activities, local physical drivers, and seagrass leaf morphology and spatial distribution (<xref ref-type="bibr" rid="B67">Unsworth et&#xa0;al., 2021</xref>).</p>
<p>Although the capacity of microplastic retention generally increases with seagrass canopy density and particle abundance, it tends to decrease with higher flow velocities, with likely varied relationships among particle types (<xref ref-type="bibr" rid="B8">Boshoff et&#xa0;al., 2023</xref>). For instance, <italic>Zostera capensis</italic> leaves are thinner and more flexible than the tougher leaves of <italic>Zostera marina</italic>, affecting their ability to trap litter (<xref ref-type="bibr" rid="B34">Kreitsberg et&#xa0;al., 2021</xref>). The significant accumulation of plastic fragments intertwined within <italic>P. oceanica</italic> seagrass beds washed ashore on Mediterranean beaches supports the idea that seagrass meadows act as effective litter traps (<xref ref-type="bibr" rid="B59">Sanchez-Vidal et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B61">Sghaier et&#xa0;al., 2025</xref>). This accumulation is likely enhanced in sheltered coastal areas and influenced by episodic events such as storms or increased runoff during heavy precipitation (<xref ref-type="bibr" rid="B30">Huang et&#xa0;al., 2020</xref>).</p>
<p>Furthermore, seagrasses directly influence sediment dynamics by reducing water velocity, which promotes sedimentation and vertical accretion (<xref ref-type="bibr" rid="B32">Jones et&#xa0;al., 2020</xref>). This sediment trapping is governed by the availability of terrestrial and marine sediments, as well as by wave and tidal energy (<xref ref-type="bibr" rid="B8">Boshoff et&#xa0;al., 2023</xref>). Vegetated coastal systems thus provide the important ecosystem service of capturing and storing environmental contaminants and organic matter, including microplastics (<xref ref-type="bibr" rid="B10">Celis-Hernandez et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B69">Veettil et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B41">Navarrete-Fern&#xe1;ndez et al., 2022</xref>). Microplastic deposition may be further enhanced by changes in relative density caused by salinity fluctuations, biofouling, and the reduced water flow within dense seagrass canopies (<xref ref-type="bibr" rid="B51">Pinheiro et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B11">Cesarini et al., 2023</xref>).</p>
<p>Aegagropiles (EGs) with fibrous aggregates of <italic>Posidonia oceanica</italic> debris and trapped sediments play a crucial ecological role as bioindicators of plastic pollution in coastal marine environments (<xref ref-type="bibr" rid="B59">Sanchez-Vidal et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B1">Alomar et&#xa0;al., 2024</xref>). Formed and deposited on shorelines, especially following storm events, EGs efficiently capture and retain microplastics and other anthropogenic pollutants, integrating contaminants over time and space (<xref ref-type="bibr" rid="B71">Verhille et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B59">Sanchez-Vidal et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B57">Rigatou et&#xa0;al., 2025</xref>). Their lignocellulosic matrix (a natural structure composed of cellulose, hemicellulose, and lignin) facilitates pollutant retention, making EGs reliable tools for detecting microplastic presence and accumulation. Variations in microplastic concentration and composition across different sites reflect local human pressures and pollution sources (<xref ref-type="bibr" rid="B59">Sanchez-Vidal et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B1">Alomar et&#xa0;al., 2024</xref>; <xref ref-type="bibr" rid="B57">Rigatou et&#xa0;al., 2025</xref>).</p>
<p>Beyond trapping pollutants, EGs provide important insights into the transport and dispersal of plastic debris within marine habitats, as they are moved by waves and currents. Their visible, persistent presence on beaches offers a practical and cost-effective method for monitoring coastal plastic pollution. Utilizing EGs as indicators supports the identification of pollution hotspots, source attribution, and evaluation of waste management effectiveness (<xref ref-type="bibr" rid="B53">Porcino et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B56">Restaino et&#xa0;al., 2023</xref>).</p>
</sec>
<sec id="s5" sec-type="conclusion">
<title>Conclusion</title>
<p>This study demonstrated that the aegagropiles (EGs), formed from the lignocellulosic debris of <italic>Posidonia oceanica</italic> meadows, can serve as effective indicators of microplastic pollution along the Tunisian coasts. Through morphological and chemical characterization of the microplastics trapped in the EGs, we found a wide diversity of plastic types of plastics often entangled with the fibrous structure of the EGs. Our findings suggest that EGs contribute to both the accumulation and transport of microplastics from shallow seagrass habitats to coastal areas, in particular during storms that facilitate their movement ashore. These results highlight the ecological role of EGs in reflecting local pollution pressures and underscore the urgent need for targeted strategies to reduce plastic input into marine environments. In particular, the high occurrence of certain polymer types points to potential land-based and maritime sources. Therefore, future research should prioritize tracing the origins of these polymers as such knowledge is critical for designing effective mitigation policies and pollution control efforts.</p>
</sec>
</body>
<back>
<sec id="s6" sec-type="data-availability">
<title>Data availability statement</title>
<p>The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.</p></sec>
<sec id="s7" sec-type="ethics-statement">
<title>Ethics statement</title>
<p>This study did not take place on any private or protected areas. No specific permissions were required for corresponding locations.</p></sec>
<sec id="s8" sec-type="author-contributions">
<title>Author contributions</title>
<p>DS: Writing &#x2013; review &amp; editing. IC: Formal Analysis, Methodology, Writing &#x2013; original draft. TB-S: Funding acquisition, Validation, Writing &#x2013; review &amp; editing. NZ: Formal Analysis, Methodology, Validation, Writing &#x2013; review &amp; editing. ME: Supervision, Validation, Writing &#x2013; review &amp; editing.</p></sec>
<sec id="s10" sec-type="COI-statement">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p></sec>
<sec id="s11" sec-type="ai-statement">
<title>Generative AI statement</title>
<p>The author(s) declare that no Generative AI was used in the creation of this manuscript.</p>
<p>Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.</p></sec>
<sec id="s12" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
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<p>Edited by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/1003141">Julius A. Ellrich</ext-link>, Alfred Wegener Institute Helmholtz Centre for Polar and Marine Research (AWI), Germany</p></fn>
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<p>Reviewed by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/3160752">Mario De Luca</ext-link>, University of Sassari, Italy; <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/3161405">Paolo Marras</ext-link>, University of Cagliari, Italy</p></fn>
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