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<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Environ. Archaeol.</journal-id>
<journal-title-group>
<journal-title>Frontiers in Environmental Archaeology</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Environ. Archaeol.</abbrev-journal-title>
</journal-title-group>
<issn pub-type="epub">2813-432X</issn>
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<publisher-name>Frontiers Media S.A.</publisher-name>
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<article-meta>
<article-id pub-id-type="doi">10.3389/fearc.2025.1626314</article-id>
<article-version article-version-type="Version of Record" vocab="NISO-RP-8-2008"/>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Brief Research Report</subject>
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</article-categories>
<title-group>
<article-title>Lab waste as hidden treasure. Early results of phytolith analysis from Iberian prehistoric post-ORA pottery powder</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" corresp="yes">
<name><surname>Ferrara</surname> <given-names>Vincenza</given-names></name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
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<uri xlink:href="https://loop.frontiersin.org/people/3061994"/>
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<contrib contrib-type="author">
<name><surname>von Allmen</surname> <given-names>Robin</given-names></name>
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<contrib contrib-type="author">
<name><surname>Salame Gonz&#x000E1;lez</surname> <given-names>Maria Fernanda</given-names></name>
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<contrib contrib-type="author">
<name><surname>Revert Franc&#x000E9;s</surname> <given-names>Elena</given-names></name>
<xref ref-type="aff" rid="aff3"><sup>3</sup></xref>
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<aff id="aff1"><label>1</label><institution>Uppsala Archeobotanical Group (UAG), Department of Archaeology, Ancient History and Conservation, Uppsala University</institution>, <city>Uppsala</city>, <country country="se">Sweden</country></aff>
<aff id="aff2"><label>2</label><institution>Department of Human Geography, Stockholm University</institution>, <city>Stockholm</city>, <country country="se">Sweden</country></aff>
<aff id="aff3"><label>3</label><institution>CRC 1070 ResourceCultures, Institute of Pre- and Early History and Archaeology of the Middle Ages and Modern Times, University of T&#x000FC;bingen</institution>, <city>T&#x000FC;bingen</city>, <country country="de">Germany</country></aff>
<author-notes>
<corresp id="c001"><label>&#x0002A;</label>Correspondence: Vincenza Ferrara, <email xlink:href="mailto:vincenza.ferrara@arkeologi.uu.se">vincenza.ferrara@arkeologi.uu.se</email></corresp>
</author-notes>
<pub-date publication-format="electronic" date-type="pub" iso-8601-date="2026-01-30">
<day>30</day>
<month>01</month>
<year>2026</year>
</pub-date>
<pub-date publication-format="electronic" date-type="collection">
<year>2025</year>
</pub-date>
<volume>4</volume>
<elocation-id>1626314</elocation-id>
<history>
<date date-type="received">
<day>23</day>
<month>05</month>
<year>2025</year>
</date>
<date date-type="rev-recd">
<day>22</day>
<month>12</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>30</day>
<month>12</month>
<year>2025</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#x000A9; 2026 Ferrara, von Allmen, Salame Gonz&#x000E1;lez and Revert Franc&#x000E9;s.</copyright-statement>
<copyright-year>2026</copyright-year>
<copyright-holder>Ferrara, von Allmen, Salame Gonz&#x000E1;lez and Revert Franc&#x000E9;s</copyright-holder>
<license>
<ali:license_ref start_date="2026-01-30">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>Initially designed to explore cultural interactions between Phoenicians and local communities in the Iberian Peninsula during the 1st millennium BCE, the work presented in our paper expanded beyond traditional Organic Residue Analysis (ORA), by attempting phytoliths extraction from post-ORA pottery powder. The aim was to assess whether such a methodological integration may enhance the understanding of past correlations between environments and material cultures in terms of pottery making and function, use of plant resources, food preparation, cultural practices, intercultural exchanges and networks. Despite working with limited sample sizes (&#x0007E;1 g per sample), phytolith analysis successfully revealed distinct environmental signatures across different archeological contexts, illustrating the valuable contribution of plant biogenic silica studies within and beyond archeological research. Furthermore, an unexpected positive result in terms of hue detection during the laboratory procedure underscores the research practice as scientific discovery process. By combining archeological, botanical, and chemical perspectives, our study showcases how phytoliths research can extend beyond traditional boundaries and complement established methodologies, reinforcing the necessity of interdisciplinary dialogue and multi-disciplinary approaches to archeology.</p></abstract>
<kwd-group>
<kwd>biogenic silica</kwd>
<kwd>blue</kwd>
<kwd>dicot phytoliths</kwd>
<kwd>Early Iron Age</kwd>
<kwd>Iberian Peninsula</kwd>
<kwd>organic residue analysis</kwd>
<kwd>Phoenicians</kwd>
<kwd>wood ash</kwd>
</kwd-group>
<funding-group>
<funding-statement>The author(s) declared that financial support was received for this work and/or its publication. This work was supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG), the Collaborative Research Centre ResourceCultures&#x02014;CRC 1070/3&#x02014;Project number 215859406, and ArchLab&#x02014;the National Infrastructure for Archaeological Science, Sweden, Swedish Research Council grant 2023-00174 (ROR id: <ext-link ext-link-type="uri" xlink:href="https://ror.org/00rky3x16">https://ror.org/00rky3x16</ext-link>).</funding-statement>
</funding-group>
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<fig-count count="4"/>
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<equation-count count="0"/>
<ref-count count="90"/>
<page-count count="12"/>
<word-count count="8938"/>
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<custom-meta-group>
<custom-meta>
<meta-name>section-at-acceptance</meta-name>
<meta-value>Archeobotany</meta-value>
</custom-meta>
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</article-meta>
</front>
<body>
<sec sec-type="intro" id="s1">
<label>1</label>
<title>Introduction</title>
<p>During the 1st millennium BCE, the Mediterranean emerged as a highly intertwined region, characterized by extensive connections, movements, exchanges of goods, plants, animals, people, and ideas. Such a dynamic interplay resulted in a flourishing and significant historical period. Human mobility, along with the cultural heritage it carried, offers an interesting perspective for studying the evolution and dynamics of past societies.</p>
<p>The people archaeologically referred to as Phoenicians originated in Canaan, a region in the eastern Mediterranean that encompasses present-day Lebanon as well as the coastal areas of Syria and Israel. They navigated across the sea to establish long-lasting settlements, where they blended with the local populations, in Sicily, Tunisia, Sardinia, Ibiza, Spain, to name a few examples (<xref ref-type="bibr" rid="B8">Aubet, 1994</xref>; <xref ref-type="bibr" rid="B24">Delgado and Ferrer, 2007</xref>; <xref ref-type="bibr" rid="B51">L&#x000F3;pez-Ruiz, 2021</xref>). The Iberian Peninsula, particularly Spain, served as a melting pot of diverse cultures and traditions, bearing significant influence from the Phoenicians, especially in the early 1st millennium BCE.</p>
<p>To explore the cultural entanglements between Phoenicians and local communities in the Iberian Peninsula during the 1st millennium BCE, within the framework of the project &#x0201C;<italic>Cultural entanglements in the Lower Guadalquivir&#x02014;Interacting ResourceCultures and socio-cultural change in the South of the Iberian Peninsula</italic>&#x0201D; (University of T&#x000FC;bingen), pottery samples were collected in 2023 for Organic Residue Analysis (ORA) from three archaeological sites in Andalusia and Extremadura (southern Iberian Peninsula) by one of the authors of this paper. The primary objective of the ORA was to identify some not-native compounds potentially linked to the Phoenicians or Mediterranean trade more in general, as well as to observe changes before and after the arrival of these settlers in the ninth century BCE.</p>
<p>Each sample weighed between 1 and 2 g and was collected from the interior surface of the pottery fragments and vessels. The analyses were conducted in the ORA laboratories at the University of T&#x000FC;bingen, and the findings are published elsewhere (<xref ref-type="bibr" rid="B73">Revert Franc&#x000E9;s, 2024</xref>; Revert Franc&#x000E9;s and Toscano, in prep<xref ref-type="fn" rid="fn0003"><sup>1</sup></xref>). However, ORA can encounter challenges in detecting plant oils, depending on the methods used (<xref ref-type="bibr" rid="B30">Drieu et al., 2025</xref>). Consequently, knowledge on plant remains from pottery residues could be limited if relying only on this method.</p>
<p>Recognizing the importance of interdisciplinarity and the value of approaching the same research question from multiple, different but integrated methodological approaches, we decided to conduct phytolith analysis on the pottery powder left from the previously done ORA process. Several authors have already shown the feasibility of phytolith analysis on sediments and/or residues from vessels and pottery, in order to obtain information on the use and cultural practices connected to those artifacts (e.g., food preparation and/or storage, <xref ref-type="bibr" rid="B44">Hart, 2011</xref>; <xref ref-type="bibr" rid="B79">Saul et al., 2013</xref>; <xref ref-type="bibr" rid="B89">Wang et al., 2022</xref>; <xref ref-type="bibr" rid="B23">Debels et al., 2024</xref>; <xref ref-type="bibr" rid="B78">Santiago-Marrero et al., 2024</xref>), and combined residue and phytolith analysis (<xref ref-type="bibr" rid="B42">Gong et al., 2025</xref>). The post-ORA samples available in the case presented here weighed only approx. 1 g, which could have represented a significant limitation for the phytolith extraction process, as it usually may require a higher amount of material depending on the context (e.g., 3.5 g, cf. <xref ref-type="bibr" rid="B55">Mazuy et al., 2024</xref>). Despite such limitations, the analysis proceeded, paving the way for novel work in the field of phytolith extraction from archaeological pottery powder after Organic Residue Analysis. Furthermore, four of the post-ORA samples were selected for pollen analysis.</p>
<p>Our paper is structured as follows. The section Data and Methods presents the archaeological context and key characteristics of the three post-ORA samples used for phytolith analysis, as well as the initial four post-ORA samples chosen for eventual integrated pollen analysis. The results obtained from our laboratory and microscopic analysis are described in the Results section, alongside the report of an unintended discovery of blue pigment in one of the samples, which underwent the first step of the pollen extraction protocol (i.e., acetolysis, cf. <xref ref-type="bibr" rid="B35">Erdtman, 1960</xref>). Our results are further discussed in the framework of the evidence gathered from the ORA analysis performed before on the same samples, and within broader considerations about how phytolith analysis, when integrated in a multi- and interdisciplinary research practice, can provide relevant insights to better understand local cultural practices.</p></sec>
<sec id="s2">
<label>2</label>
<title>Data and methods</title>
<p>Phytolith analysis was conducted on three samples made of post-ORA pottery powder (<xref ref-type="table" rid="T1">Table 1</xref>) from the archaeological sites of Pe&#x000F1;alosa, Tejada la Vieja, and Hacienda, located in the modern-day province of Huelva, in southwestern Spain (<xref ref-type="fig" rid="F1">Figure 1</xref>).</p>
<table-wrap position="float" id="T1">
<label>Table 1</label>
<caption><p>Samples for phytolith analysis.</p></caption>
<table frame="box" rules="all">
<thead>
<tr>
<th valign="top" align="left"><bold>Name</bold></th>
<th valign="top" align="center"><bold>Weight of initial powder (g)</bold></th>
<th valign="top" align="center"><bold>Sampled part of the vessel</bold></th>
<th valign="top" align="center"><bold>Site</bold></th>
<th valign="top" align="center"><bold>Chronology</bold></th>
<th valign="top" align="center"><bold>Weight of final sediment after phytolith extraction (g)</bold></th>
</tr>
</thead>
<tbody>
<tr>
<td valign="top" align="left">PE08</td>
<td valign="top" align="center">1.003</td>
<td valign="top" align="center">Body sherd of a small burnished patera</td>
<td valign="top" align="center">Pe&#x000F1;alosa</td>
<td valign="top" align="center">ca. 12th&#x02212;8th cent. BCE</td>
<td valign="top" align="center">0.015</td>
</tr>
<tr>
<td valign="top" align="left">TV003</td>
<td valign="top" align="center">1.048</td>
<td valign="top" align="center">Rim sherd of a coarse storage jar</td>
<td valign="top" align="center">Tejada la Vieja</td>
<td valign="top" align="center">9th&#x02212;4th cent. BCE</td>
<td valign="top" align="center">0.010</td>
</tr>
<tr>
<td valign="top" align="left">HA002</td>
<td valign="top" align="center">1.076</td>
<td valign="top" align="center">Upper part of a coarse storage jar with one handle</td>
<td valign="top" align="center">Hacienda</td>
<td valign="top" align="center">8th&#x02212;6th cent. BCE</td>
<td valign="top" align="center">0.053</td>
</tr></tbody>
</table>
</table-wrap>
<fig position="float" id="F1">
<label>Figure 1</label>
<caption><p>Map of the southwest of the Iberian Peninsula with the archeological sites named in the map. The drawing of the Lacus Ligustinus is based on the research of Oswaldo Arteaga (<xref ref-type="bibr" rid="B7">Arteaga and Roos, 1992</xref>; <xref ref-type="bibr" rid="B6">Arteaga et al., 2016</xref>).</p></caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fearc-04-1626314-g0001.tif">
<alt-text content-type="machine-generated">Map of the Iberian Peninsula with highlighted archeological sites including Tejada la Vieja, Pe&#x000F1;alosa, Hacienda, Siete Arroyos, and El Turunuelo de Guare&#x000F1;a. An inset shows the location within Europe. A scale indicates a distance of two hundred kilometers.</alt-text>
</graphic>
</fig>
<p>Pe&#x000F1;alosa (excavated in 1990) is the oldest of these sites, primarily dating to the Late Bronze Age, with an earlier, less-documented occupation during the Chalcolithic period. The excavated area of this settlement, featuring six pit houses, contained typical Late Bronze Age pottery, with the exception of a small fragment of wheel-made Phoenician red-slipped and burnished ware. Excluding this fragment, the assemblage could confidently be dated to the last centuries of the 2nd millennium BCE. However, the presence of this fragment, along with silver slag, prompted excavators to assign a date to the settlement in the ninth to eighth centuries BCE (<xref ref-type="bibr" rid="B39">Garc&#x000ED;a Sanz and Fern&#x000E1;ndez Jurado, 2000</xref>). The possibility of the Phoenician fragment being an intrusive artifact cannot be entirely ruled out. Considering this and the otherwise uniform characteristics of Pe&#x000F1;alosa, a dating range between the twelfth to eighth centuries is proposed here. From this site, phytolith analysis was performed on 1.003 g of post-ORA pottery powder recovered from the inner wall of a body sherd of a small burnished patera (i.e., bowl; <bold>sample PE008</bold>).</p>
<p>Tejada la Vieja is a fortified settlement, with its defensive walls dating to the late ninth century BCE. Covering approximately 6.4 ha, almost the entire area appears to have been densely constructed. The settlement seems to have been abandoned in the fourth century BCE. It is possible that the establishment of Tejada la Vieja is linked to the abandonment of Pe&#x000F1;alosa, situated merely 3 km away (<xref ref-type="bibr" rid="B36">Fern&#x000E1;ndez Jurado, 1987</xref>). Phytolith analysis was done on 1.048 g of post-ORA pottery powder from the inner wall of a rim sherd of a coarse storage jar, found in an archeological excavation in 1981 (<bold>sample TV003</bold>).</p>
<p>The Hacienda site (excavated in 2023) is located beneath the old Treasury Building in the city of Huelva. Recent renovation works on the building facilitated archaeological excavations, uncovering remains of the city&#x00027;s harbor dating back to the Early Iron Age. The excavation and analysis of the recovered materials are still in progress. 1.076 g of post-ORA pottery powder from the inner wall of the upper part of a coarse storage jar with one handle was analyzed for phytolith content (<bold>sample HA002</bold>).</p>
<p>Samples for ORA analysis, conducted in the ORA laboratories at the University of T&#x000FC;bingen, were extracted by one of the authors of this paper (<xref ref-type="bibr" rid="B73">Revert Franc&#x000E9;s, 2024</xref>; Revert Franc&#x000E9;s and Toscano, in prep (see text footnote <xref ref-type="fn" rid="fn0003">1</xref>)) in 2023 from the inner surface of the sherds using a drill, according to a standard method (<xref ref-type="bibr" rid="B46">Heron et al., 1991</xref>; <xref ref-type="bibr" rid="B20">Charters et al., 1993</xref>; <xref ref-type="bibr" rid="B72">Regert, 2011</xref>). ORA sampling method foresees the removal of a first layer of material from the artifact, due to potential contamination from soil and/or the handling process after excavation, and the use for Organic Residue Analysis of only the powder obtained from drilling of the subsequent layer, located just below the first (e.g., <xref ref-type="bibr" rid="B71">Rageot et al., 2019</xref>; <xref ref-type="bibr" rid="B80">Spiteri et al., 2025</xref>).</p>
<p>After ORA analysis, the same samples, with identical weight, were brought in 2024 at the Paleobiology Laboratory (Department of Earth Sciences, Uppsala University) to attempt phytolith analysis by the Uppsala Archeobotanical Group on these post-ORA pottery samples.</p>
<p>Typically, ORA samples weigh between 1 and 2 g. When sampling for ORA, to prevent further damage to the ceramics, no surplus material was collected for potential integrative analyses. Consequently, in the case presented in this paper, we worked with particularly small samples suitable for phytolith analysis. Moreover, such a low amount of post-ORA pottery powder available per sample (approx. 1 g) could not allow to conduct both phytolith and pollen analysis on the same material. For such a reason, we selected four new post-ORA pottery residue samples for an initially planned pollen analysis (<xref ref-type="table" rid="T2">Table 2</xref>). These samples come from the archeological sites of Siete Arroyos (Seville), Hacienda (Huelva), and Casas del Turu&#x000F1;uelo de Guare&#x000F1;a (Badajoz), in southwestern Spain.</p>
<table-wrap position="float" id="T2">
<label>Table 2</label>
<caption><p>Samples for intended pollen analysis.</p></caption>
<table frame="box" rules="all">
<thead>
<tr>
<th valign="top" align="left"><bold>Name</bold></th>
<th valign="top" align="center"><bold>Weight of initial powder (g)</bold></th>
<th valign="top" align="center"><bold>Sampled part of the vessel</bold></th>
<th valign="top" align="center"><bold>Site</bold></th>
<th valign="top" align="center"><bold>Chronology</bold></th>
</tr>
</thead>
<tbody>
<tr>
<td valign="top" align="left">SA002_L2</td>
<td valign="top" align="center">1.000</td>
<td valign="top" align="center">Bottom of a coarse pottery bottle</td>
<td valign="top" align="center">Siete Arroyos</td>
<td valign="top" align="center">19th&#x02212;17th cent. BCE</td>
</tr>
<tr>
<td valign="top" align="left">SA_Bod003</td>
<td valign="top" align="center">1.002</td>
<td valign="top" align="center">Soil sample from inside the vessel SA002_L2</td>
<td valign="top" align="center">Siete Arroyos</td>
<td valign="top" align="center">19th&#x02212;17th cent. BCE</td>
</tr>
<tr>
<td valign="top" align="left">HA012</td>
<td valign="top" align="center">1.019</td>
<td valign="top" align="center">Bottom of a coarse storage jar</td>
<td valign="top" align="center">Hacienda</td>
<td valign="top" align="center">8th&#x02212;6th cent. BCE</td>
</tr>
<tr>
<td valign="top" align="left">TR016</td>
<td valign="top" align="center">1.011</td>
<td valign="top" align="center">Body sherd of a coarse vessel</td>
<td valign="top" align="center">El Turu&#x000F1;uelo</td>
<td valign="top" align="center">5th cent. BCE</td>
</tr></tbody>
</table>
</table-wrap>
<p>Siete Arroyos is a burial site dating to the Early Bronze Age. The excavation and analysis of the archeological materials are ongoing (<xref ref-type="bibr" rid="B14">Bartelheim et al., 2025</xref>). From this site, pottery powder from a vessel and soil from its interior were sampled.</p>
<p>The excavation of the archeological site of Casas del Turu&#x000F1;uelo de Guare&#x000F1;a (hereafter referred to as El Turu&#x000F1;uelo) is currently underway and is expected to continue in the coming years. The site presents a complex archeological narrative: evidence has been found of a banquet, the sacrificial slaughter of 41 equids alongside other animals such as cattle and pigs, and a deliberate conflagration that abruptly ended its use (<xref ref-type="bibr" rid="B74">Rodr&#x000ED;guez Gonz&#x000E1;lez and Celestino, 2017</xref>, <xref ref-type="bibr" rid="B75">2019</xref>; <xref ref-type="bibr" rid="B47">Ibora Eres et al., 2023</xref>).</p>
<p>Phytolith extraction was conducted at the Paleobiology Laboratory (Department of Earth Sciences, Uppsala University), according to <xref ref-type="bibr" rid="B55">Mazuy et al. (2024)</xref> protocol, which allows the extraction of phytoliths from samples characterized by low biogenic silica content. This protocol foresees the deflocculation of the samples with magnetic stirring and the separation of its fine fraction via sieving (200 &#x003BC;m). Carbonates are then removed by hydrochloric acid treatment and organic matter with potassium hydroxide in a hot bath. In the resulting solution, phytoliths are then extracted by heavy liquid flotation and further cleaned from remaining organic matter through boiling in hydrogen peroxide.</p>
<p>The phytoliths resulting from the extraction were identified, counted and interpreted at the Department of Archaeology, Ancient History and Conservation (Uppsala University), using a light microscope at &#x000D7; 400 magnification. A minimum of 200 diagnostic morphotypes were identified and categorized into plant taxonomic groups according to the International Code for Phytolith Nomenclature (ICPN; <xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>) and the PhytCore online database (<xref ref-type="bibr" rid="B3">Albert et al., 2016</xref>). For each sample, we calculated each morphotype&#x00027;s relative abundance and analyzed their assemblages in terms of the ratio of inflorescence to culm-leaves morphotypes, long to short cell phytoliths, Dicotyledonous to Poaceae morphotypes (cf. <xref ref-type="bibr" rid="B37">Ferrara, 2024</xref> for details about how to calculate these indices).</p>
<p>The ratio of inflorescence to culm-leaves phytoliths indicates the amount of <sc>Elongate dendritic</sc> phytoliths (from inflorescence parts of grasses) to <sc>Elongate entire</sc> and <sc>Elongate sinuate</sc> (from culms and leaves; <xref ref-type="bibr" rid="B64">Piperno, 1988</xref>; <xref ref-type="bibr" rid="B85">Tsartsidou et al., 2007</xref>; <xref ref-type="bibr" rid="B27">Delhon et al., 2020</xref>), and can be informative about the presence of spikelets and/or straws in an assemblage. <sc>Elongate</sc> phytoliths, produced by grasses from the Poaceae family (<xref ref-type="bibr" rid="B86">Twiss et al., 1969</xref>; <xref ref-type="bibr" rid="B38">Fredlund and Tieszen, 1994</xref>; <xref ref-type="bibr" rid="B64">Piperno, 1988</xref>; <xref ref-type="bibr" rid="B10">Ball et al., 2001</xref>; <xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>), can be distinguished among <sc>Elongate entire</sc> morphotype from the stems and <sc>Elongate sinuate</sc> from the leaves, while <sc>Elongate dendritic</sc> from the inflorescence of the plant (<xref ref-type="bibr" rid="B76">Rosen, 1992</xref>; <xref ref-type="bibr" rid="B9">Ball et al., 1999</xref>; <xref ref-type="bibr" rid="B68">Portillo et al., 2006</xref>; <xref ref-type="bibr" rid="B4">Albert et al., 2008</xref>).</p>
<p>The ratio of long cell to short cell phytoliths can provide information on the grass composition in terms of age and phenological phase, thus we can infer if local vegetation was formed by young or mature grasses (<xref ref-type="bibr" rid="B28">Delhon et al., 2024</xref>). Furthermore, when it comes to assemblage analysis of grass short cell phytoliths, it is possible to distinguish among the following three Poaceae subfamilies, represented by these specific phytolith morphotypes: <sc>Rondel</sc> (<xref ref-type="bibr" rid="B86">Twiss et al., 1969</xref>; <xref ref-type="bibr" rid="B38">Fredlund and Tieszen, 1994</xref>; <xref ref-type="bibr" rid="B66">Piperno and Pearsall, 1998</xref>; <xref ref-type="bibr" rid="B12">Barboni and Bremond, 2009</xref>), <sc>Trapezoid</sc> (<xref ref-type="bibr" rid="B12">Barboni and Bremond, 2009</xref>) and <sc>Crenate</sc> (<xref ref-type="bibr" rid="B86">Twiss et al., 1969</xref>; <xref ref-type="bibr" rid="B38">Fredlund and Tieszen, 1994</xref>; <xref ref-type="bibr" rid="B13">Barboni et al., 2007</xref>) as representative of Pooideae (C3 grasses growing in temperate climates); <sc>Saddle</sc> (<xref ref-type="bibr" rid="B65">Piperno, 2006</xref>; <xref ref-type="bibr" rid="B54">Madella et al., 2016</xref>) and <sc>Rondel</sc> in association with them (<xref ref-type="bibr" rid="B11">Bamford et al., 2006</xref>; <xref ref-type="bibr" rid="B12">Barboni and Bremond, 2009</xref>) as indicators of Chloridoideae (C4 grasses in dry and warm environments); <sc>Bilobate</sc> (<xref ref-type="bibr" rid="B86">Twiss et al., 1969</xref>; <xref ref-type="bibr" rid="B38">Fredlund and Tieszen, 1994</xref>; <xref ref-type="bibr" rid="B12">Barboni and Bremond, 2009</xref>), <sc>Polylobate</sc> (<xref ref-type="bibr" rid="B86">Twiss et al., 1969</xref>; <xref ref-type="bibr" rid="B38">Fredlund and Tieszen, 1994</xref>; <xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>) and <sc>Cross</sc> indicating usually Panicoideae (C4 grasses in warm and wet contexts).</p>
<p>Comparing Dicots vs. Poaceae morphotypes can inform on the different origins of the phytoliths (trees or shrubs vs. grasses). The following morphotypes are attributable to dicotyledonous: <sc>Spheroid</sc> (<xref ref-type="bibr" rid="B16">Bozarth, 1992</xref>; <xref ref-type="bibr" rid="B5">Alexandre et al., 1997</xref>; <xref ref-type="bibr" rid="B2">Albert et al., 1999</xref>; <xref ref-type="bibr" rid="B77">Runge, 1999</xref>; <xref ref-type="bibr" rid="B26">Delhon et al., 2003</xref>) and <sc>Blocky</sc> (<xref ref-type="bibr" rid="B84">Tsartsidou et al., 2015</xref>; <xref ref-type="bibr" rid="B15">Boixadera et al., 2016</xref>; <xref ref-type="bibr" rid="B58">Ntinou and Tsartsidou, 2017</xref>; <xref ref-type="bibr" rid="B18">Burguet-Coca et al., 2020</xref>; <xref ref-type="bibr" rid="B49">Kraushaar et al., 2021</xref>; <xref ref-type="bibr" rid="B83">Tencariu et al., 2022</xref>).</p>
<p>The chemical preparation of samples to extract pollen was performed according to the acetolysis protocol (<xref ref-type="bibr" rid="B35">Erdtman, 1960</xref>), which is the standard chemical treatment to enhance pollen morphological features and remove non-pollen organic matter. Acetolyzed samples were analyzed under a bright-field microscope, Nikon Eclipse Ni, with a 40 &#x000D7; objective.</p></sec>
<sec sec-type="results" id="s3">
<label>3</label>
<title>Results</title>
<p>Phytoliths were present in all the three samples and in good preservation state, despite the initial low amount of pottery powder processed (<xref ref-type="fig" rid="F2">Figure 2</xref>). In all the three samples, there is a predominance of short cell vs. long cell phytoliths (PE08 53.5% vs. 21%; TV003 45% vs. 36%; HA002 64.5% vs. 18%). All the three samples show, however, overall high percentages of dicot phytoliths as well: PE08 14.5%, TV003 10%, HA002 10.5% (<xref ref-type="table" rid="T3">Table 3</xref>), reflected by the Dicotyledonous/Poaceae index values (<xref ref-type="table" rid="T4">Table 4</xref>), thus attesting the presence of residues from dicot plants.</p>
<fig position="float" id="F2">
<label>Figure 2</label>
<caption><p>Examples of phytoliths extracted from the samples: <bold>(a)</bold> <sc>Blocky</sc> (PE08); <bold>(b)</bold> <sc>Blocky</sc>, side view (PE08); <bold>(c)</bold> <sc>Elongate dendritic</sc> (TV003); <bold>(d)</bold> <sc>Elongate dendritic</sc>, side view (TV003); <bold>(e)</bold> <sc>Elongate entire</sc> (HA002); <bold>(f)</bold> <sc>Polylobate</sc> (HA002); <bold>(g)</bold> <sc>Rondel</sc> (TV003); <bold>(h)</bold> <sc>Spheroid psilate</sc> (TV003).</p></caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fearc-04-1626314-g0002.tif">
<alt-text content-type="machine-generated">Microscopic images labeled a to h, each showing different small silica particles under magnification, with a scale of fifty micrometers provided in each panel. The particles vary in shape and transparency, some more defined and angular like in panels a, b, and e, while others are irregular or faintly visible like in panels c, d, f, g, and h.</alt-text>
</graphic>
</fig>
<table-wrap position="float" id="T3">
<label>Table 3</label>
<caption><p>Phytolith morphotypes: absolute counts and frequency (%) per sample (PE08&#x02014;TV003&#x02014;HA002).</p></caption>
<table frame="box" rules="all">
<thead>
<tr>
<th/>
<th valign="top" align="center" colspan="2"><bold>PE08</bold></th>
<th valign="top" align="center" colspan="2"><bold>TV003</bold></th>
<th valign="top" align="center" colspan="2"><bold>HA002</bold></th>
</tr>
<tr>
<th valign="top" align="center"><bold>Phytolith morphotype</bold></th>
<th valign="top" align="center"><bold>Absolute count</bold></th>
<th valign="top" align="center"><bold>Frequency (%)</bold></th>
<th valign="top" align="center"><bold>Absolute count</bold></th>
<th valign="top" align="center"><bold>Frequency (%)</bold></th>
<th valign="top" align="center"><bold>Absolute count</bold></th>
<th valign="top" align="center"><bold>Frequency (%)</bold></th>
</tr>
</thead>
<tbody>
<tr>
<td valign="top" align="left"><sc>Spheroid psilate</sc></td>
<td valign="top" align="center">7</td>
<td valign="top" align="center">3.5</td>
<td valign="top" align="center">2</td>
<td valign="top" align="center">1</td>
<td valign="top" align="center">9</td>
<td valign="top" align="center">4.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Spheroid echinate</sc></td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">1</td>
<td valign="top" align="center">0.5</td>
<td valign="top" align="center">1</td>
<td valign="top" align="center">0.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Acute bulbosus</sc></td>
<td valign="top" align="center">6</td>
<td valign="top" align="center">3</td>
<td valign="top" align="center">5</td>
<td valign="top" align="center">2.5</td>
<td valign="top" align="center">9</td>
<td valign="top" align="center">4.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Blocky</sc></td>
<td valign="top" align="center">22</td>
<td valign="top" align="center">11</td>
<td valign="top" align="center">17</td>
<td valign="top" align="center">8.5</td>
<td valign="top" align="center">11</td>
<td valign="top" align="center">5.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Bulliform flabellate</sc></td>
<td valign="top" align="center">16</td>
<td valign="top" align="center">8</td>
<td valign="top" align="center">13</td>
<td valign="top" align="center">6.5</td>
<td valign="top" align="center">4</td>
<td valign="top" align="center">2</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Elongate entire</sc></td>
<td valign="top" align="center">33</td>
<td valign="top" align="center">16.5</td>
<td valign="top" align="center">57</td>
<td valign="top" align="center">28.5</td>
<td valign="top" align="center">33</td>
<td valign="top" align="center">16.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Elongate sinuate</sc></td>
<td valign="top" align="center">5</td>
<td valign="top" align="center">2.5</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Elongate dendritic</sc></td>
<td valign="top" align="center">4</td>
<td valign="top" align="center">2</td>
<td valign="top" align="center">15</td>
<td valign="top" align="center">7.5</td>
<td valign="top" align="center">3</td>
<td valign="top" align="center">1.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Bilobate</sc></td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">3</td>
<td valign="top" align="center">1.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Polylobate</sc></td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">1</td>
<td valign="top" align="center">0.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Crenate</sc></td>
<td valign="top" align="center">1</td>
<td valign="top" align="center">0.5</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">0</td>
<td valign="top" align="center">13</td>
<td valign="top" align="center">6.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Rondel</sc></td>
<td valign="top" align="center">82</td>
<td valign="top" align="center">41</td>
<td valign="top" align="center">83</td>
<td valign="top" align="center">41.5</td>
<td valign="top" align="center">92</td>
<td valign="top" align="center">46</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Trapezoid</sc></td>
<td valign="top" align="center">24</td>
<td valign="top" align="center">12</td>
<td valign="top" align="center">7</td>
<td valign="top" align="center">3.5</td>
<td valign="top" align="center">21</td>
<td valign="top" align="center">10.5</td>
</tr>
<tr>
<td valign="top" align="left"><bold>Total</bold></td>
<td valign="top" align="center"><bold>200</bold></td>
<td valign="top" align="center"><bold>100</bold></td>
<td valign="top" align="center"><bold>200</bold></td>
<td valign="top" align="center"><bold>100</bold></td>
<td valign="top" align="center"><bold>200</bold></td>
<td valign="top" align="center"><bold>100</bold></td>
</tr>
</tbody>
</table>
</table-wrap>
<table-wrap position="float" id="T4">
<label>Table 4</label>
<caption><p>Phytolith morphotypes assemblage analysis per sample (PE08&#x02014;TV003&#x02014;HA002).</p></caption>
<table frame="box" rules="all">
<thead>
<tr>
<th valign="top" align="left"><bold>Assemblage analysis (morphotypes ratio)</bold></th>
<th valign="top" align="center"><bold>PE08</bold></th>
<th valign="top" align="center"><bold>TV003</bold></th>
<th valign="top" align="center"><bold>HA002</bold></th>
</tr>
</thead>
<tbody>
<tr>
<td valign="top" align="left">long cell</td>
<td valign="top" align="center">21</td>
<td valign="top" align="center">36</td>
<td valign="top" align="center">18</td>
</tr>
<tr>
<td valign="top" align="left">short cell</td>
<td valign="top" align="center">53.5</td>
<td valign="top" align="center">45</td>
<td valign="top" align="center">64.5</td>
</tr>
<tr>
<td valign="top" align="left"><bold>Index long cell/short cell</bold></td>
<td valign="top" align="center"><bold>0.4</bold></td>
<td valign="top" align="center"><bold>0.8</bold></td>
<td valign="top" align="center"><bold>0.3</bold></td>
</tr>
<tr>
<td valign="top" align="left">DICOTS</td>
<td valign="top" align="center">14.5</td>
<td valign="top" align="center">10</td>
<td valign="top" align="center">10.5</td>
</tr>
<tr>
<td valign="top" align="left">POACEAE</td>
<td valign="top" align="center">53.5</td>
<td valign="top" align="center">45</td>
<td valign="top" align="center">64.5</td>
</tr>
<tr>
<td valign="top" align="left"><bold>Index D/P</bold></td>
<td valign="top" align="center"><bold>0.3</bold></td>
<td valign="top" align="center"><bold>0.2</bold></td>
<td valign="top" align="center"><bold>0.2</bold></td>
</tr>
<tr>
<td valign="top" align="left"><sc>Elongate dendritic</sc></td>
<td valign="top" align="center">2</td>
<td valign="top" align="center">7.5</td>
<td valign="top" align="center">1.5</td>
</tr>
<tr>
<td valign="top" align="left"><sc>Elongate entire</sc> &#x0002B; <sc>Elongate sinuate</sc></td>
<td valign="top" align="center">19</td>
<td valign="top" align="center">28.5</td>
<td valign="top" align="center">16.5</td>
</tr>
<tr>
<td valign="top" align="left"><bold>Index inflorescence/culm-leaves</bold></td>
<td valign="top" align="center"><bold>0.11</bold></td>
<td valign="top" align="center"><bold>0.26</bold></td>
<td valign="top" align="center"><bold>0.09</bold></td>
</tr></tbody>
</table>
</table-wrap>
<p>If the three samples show similarities in such respect, there are important differences among them.</p>
<p>HA002 is the sample with the highest number of short cells (64.5%) when compared to the others, and these include also phytolith morphotypes attributable to C4 Panicoid grasses (<sc>Bilobate</sc> 1.5% and <sc>Polylobate</sc> 0.5%; cf. <xref ref-type="bibr" rid="B59">Out and Madella, 2016</xref>). HA002 also has a slight presence of <sc>Spheroid echinate</sc> phytoliths (0.5%), attributable to palm trees (<xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>, 195&#x02013;196).</p>
<p>On the contrary, TV003 is the sample that presents the highest percentage of long cell phytoliths among the three, 36%, of which 7.5% is formed by <sc>Elongate dendritic</sc> morphotypes (produced in the inflorescence parts of a plant and/or in crop by-product, cf. <xref ref-type="bibr" rid="B78">Santiago-Marrero et al., 2024</xref>). Moreover, when this morphotype has been identified at the microscope and categorized, it was possible to attest that the recognized <sc>Elongate dendritic</sc> phytoliths (see example in <xref ref-type="fig" rid="F2">Figure 2</xref>) belonged to C3 wild grasses and/or cereals and not to Panicoids (<italic>sensu</italic> <xref ref-type="bibr" rid="B9">Ball et al., 1999</xref>; <xref ref-type="bibr" rid="B52">Lu et al., 2009</xref>; <xref ref-type="bibr" rid="B53">Madella et al., 2014</xref>; <xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>). <sc>Elongated dendritic</sc> phytoliths have not been found in such high percentages in the other two samples (PE08 2%; HA002 1.5%). The phytolith assemblage of TV003 contains a small percentage of <sc>Spheroid echinate</sc> as well (0.5%).</p>
<p>Phytolith morphotypes percentages and assemblage analysis are presented for all the three samples in <xref ref-type="table" rid="T3">Tables 3</xref>, <xref ref-type="table" rid="T4">4</xref>, and visually shown in <xref ref-type="fig" rid="F3">Figure 3</xref>.</p>
<fig position="float" id="F3">
<label>Figure 3</label>
<caption><p>Phytolith morphotypes percentages in the three samples PE08, TV003, and HA002.</p></caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fearc-04-1626314-g0003.tif">
<alt-text content-type="machine-generated">Three pie charts labeled PE08, TV003, and HA002 compare different phytoliths categories. Each chart displays percentage values with distinct colors for categories.</alt-text>
</graphic>
</fig>
<p>Results from pollen analysis did not attest the presence of pollen in the samples analyzed. However, after the first preprocessing step of the pollen acetolysis protocol (water bath 90 &#x000B0;C with Sodium hydroxide 10% for 5 min), the sample TR016 showed the presence of a vivid blue compound (7.5B7/8 or 7.5B8/8; <xref ref-type="fig" rid="F4">Figure 4</xref>).</p>
<fig position="float" id="F4">
<label>Figure 4</label>
<caption><p>Vivid blue compound present in sample TR016 after the first preprocessing step of pollen acetolysis.</p></caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fearc-04-1626314-g0004.tif">
<alt-text content-type="machine-generated">Four centrifuge tubes containing different colored liquids are placed in a white plastic rack. The first tube has a blue-colored liquid.</alt-text>
</graphic>
</fig>
</sec>
<sec sec-type="discussion" id="s4">
<label>4</label>
<title>Discussion</title>
<p>Evidence from phytolith analysis provides integrative information to the ORA results that, when interpreted within the broader archeological context of the pottery remains analyzed, can provide new insights into the local environment, plant use in terms of dietary, cultural and symbolic practices, allowing us also to make assumptions about intercultural influence and exchange.</p>
<p>Despite the modest number of samples, within a comparative perspective, the results from phytolith analysis show a high amount of dicot phytolith morphotypes from wood and bark in all the three samples.</p>
<p>Even relatively modest percentages of dicots phytolith morphotypes can be interpreted as meaningful (<xref ref-type="bibr" rid="B19">Carnelli et al., 2004</xref>; <xref ref-type="bibr" rid="B85">Tsartsidou et al., 2007</xref>), as dicots have a low phytoliths production compared to grasses. Their high number in all the three samples (each of them located in diverse archeological contexts) can thus be interpreted beyond any doubt (cf. <xref ref-type="bibr" rid="B89">Wang et al., 2022</xref> for a similar argument about the interpretation of a high amount of grass phytoliths extracted from pottery residues). First and foremost, such a high amount of dicot phytoliths (when associated with higher percentages of C3 grasses, as in the samples analyzed here) can inform about the local paleoenvironment of the time, characterized by a general temperate climate and related type of vegetation (<xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>). Furthermore, the fact that such a high amount of dicots phytoliths from wood and bark is present in all the three structurally diverse and geographically-chronologically distinct pottery fragments (a small burnished patera&#x02014;Pe&#x000F1;alosa PE08, a rim sherd of a coarse storage jar&#x02014;Tejada TV003 and an upper part of a coarse storage jar with one handle&#x02014;Hacienda HA002) can be interpreted as a form of practice common to all the three different sites. One first hypothesis is that such a common practice may have been either local indigenous or brought from outside in the past, and already well-diffused and adopted by that time. Since the three samples belong to three pots different in both size and specific shape, one potential explanation for their common high amount of dicot phytoliths could be that these phytoliths are evidence of wood ashes use as pottery temper or to improve the quality of the clay (<xref ref-type="bibr" rid="B22">Coria-Noguera et al., 2021</xref>; <xref ref-type="bibr" rid="B40">Gardner et al., 2025</xref>).</p>
<p>Another possible explanation for the origin of the counted dicot phytoliths might be rituals of light burning of small wood branches to produce smoke and/or fragrance (<italic>sensu</italic> <xref ref-type="bibr" rid="B17">Buonasera et al., 2023</xref>). However, this last explanation would apply only to the small patera&#x02014;PE008, since the other two fragments derive from bigger pots that could not have had such a use.</p>
<p>Preserved in the fabric of pottery (<xref ref-type="bibr" rid="B88">Wallis et al., 2014</xref>; <xref ref-type="bibr" rid="B34">Dzhanfezova, 2021</xref>), when not in a forest context, higher proportions of dicot phytoliths (especially those derived from wood/bark) have been associated in the literature with ash presence (<xref ref-type="bibr" rid="B25">Delhon, 2010</xref>; <xref ref-type="bibr" rid="B32">Dudgeon, 2024</xref>). As demonstrated by <xref ref-type="bibr" rid="B1">Albert et al. (2003)</xref>, phytoliths from wood ashes are highly informative in archeological contexts and <xref ref-type="bibr" rid="B25">Delhon (2010)</xref> observed how this type of phytoliths has no specific characteristics indicative of their exposure to high heat (i.e., their supposed blackish color as charred phytoliths). Consequently, the high percentage of dicot phytoliths in all the three samples investigated in this paper could be correlated with the presence of wood ash in the pottery fabric.</p>
<p>One last alternative explanation that might explain such a large quantity of dicot phytoliths in all the three fragments can be the use of wooden tools as part of food processing, for example, to grind or crush plant materials (<xref ref-type="bibr" rid="B70">Portillo et al., 2017</xref>, <xref ref-type="bibr" rid="B69">2024</xref>). However, such a hypothetical explanation does not take into consideration the distinctive dimensions and structural characteristics of the three vessels, which differ significantly in size, shape, and&#x02014;likely&#x02014;function. Consequently, the probability of an identical use-related phytolith signature across all three samples appears highly unlikely.</p>
<p>We conclude that the most probable explanation of the high number of dicot phytoliths in all the three fragments may be indicative of wood-ash tempering practices of the pottery or its clay improvement; practices that, moreover, appear to be connected with the local culture at the broader regional level (<xref ref-type="bibr" rid="B22">Coria-Noguera et al., 2021</xref>). Another element of commonality among the three samples is indicated by the ORA results, which show the presence of beeswax and conifer resin. According to the literature and the specific use we have hypothesized for every single pot, as explained further below, the mixture of beeswax and conifer resin could be interpreted as mastic used for sealing and insulation purposes (<xref ref-type="bibr" rid="B43">Grace, 1996</xref>).</p>
<p>The comparison of phytolith analysis and ORA results for every single sample brings further integrative insights as well.</p>
<p>The results of the solvent extraction of lipids from sample <bold>PE08</bold>, published in <xref ref-type="bibr" rid="B73">Revert Franc&#x000E9;s (2024)</xref>, show well-preserved beeswax, with a concentration of 334 &#x003BC;g/g of pottery powder. Additionally, a malic acid-rich fruit product was detected, with a concentration of 2 &#x003BC;g/g of pottery powder. Due to the elevated presence of beeswax, we then hypothesize that this small patera may have been used as a lamp (according to the interpretation of <xref ref-type="bibr" rid="B73">Revert Franc&#x000E9;s, 2024</xref>) or to contain some sort of products (e.g., cosmetic scented balms made with natural ingredients, probably fruits), while we exclude the ritual use to create light fragrance smoking (as advanced above, when providing a second interpretative option that could explain the high presence of dicot phytoliths from wood and bark).</p>
<p>In sample <bold>TV003</bold> (rim sherd of a coarse storage jar), the high percentage of long cell phytoliths (36%), of which 7.5% <sc>Elongate dendritic</sc> from C3 wild grasses or cereals (<xref ref-type="bibr" rid="B9">Ball et al., 1999</xref>; <xref ref-type="bibr" rid="B52">Lu et al., 2009</xref>; <xref ref-type="bibr" rid="B53">Madella et al., 2014</xref>; <xref ref-type="bibr" rid="B57">Neumann et al., 2019</xref>), may attest the use of the vessel for storage of adult plants and/or crop by-products. The evidence of C3 cereals from the phytolith assemblages may be indicative of their cultivation and use at that time in the area, as reported by other sources (cf. <xref ref-type="bibr" rid="B60">P&#x000E9;rez-Jord&#x000E0;, 2020</xref>). From the ORA, a probable presence of ruminant fat, at an exceptionally high concentration of 5,637 &#x003BC;g/g of pottery powder was identified, which could have been used as a preservative for the adult plants and/or crop by-products processed or cooked in some way. A malic acid-rich fruit product (17 &#x003BC;g/g of pottery powder) completed the assemblage of organic compounds found in this coarse jar, whose most probable use could have thus been storage of prepared/processed food of some sort.</p>
<p>Sample <bold>HA002</bold> is derived from the upper part of a coarse jar with one handle. The potential interpretation of its scope as a serving jar is supported by both ORA and phytoliths results. From the ORA, an interesting and well-preserved mixture (470 &#x003BC;g/g of pottery powder) was identified, consisting of animal fat, conifer resin, other plant products (high degraded triterpenes) and wine (tartaric acid: 2.27 &#x003BC;g/g of pottery powder; malic acid: 0.77 &#x003BC;g/g of pottery powder; %TA: 74%, following <xref ref-type="bibr" rid="B31">Drieu et al., 2021</xref>), suggesting multiple possible liquid contents for this vessel. In addition, the presence of <sc>Spheroid echinate</sc> phytoliths in this sample may be evidence of the palm tree <italic>Chamaerops humilis</italic>, endemic of the Iberian Peninsula and widespread in the site area. Such an assumption is corroborated by the ORA results, which indicate the presence of <italic>Chamaerops humilis</italic> as well, through evidence of triterpenes (which are contained in the palm tree, cf. <xref ref-type="bibr" rid="B82">Taibi et al., 2025</xref>). <xref ref-type="bibr" rid="B23">Debels et al. (2024)</xref> do not exclude the idea that phytoliths from palm trees could be indicative of certain food products (e.g., palm oil and palm wine, commonly prepared at the time in the area according to <xref ref-type="bibr" rid="B62">P&#x000E9;rez-Jord&#x000E0; et al., 2017</xref>). However, since the ORA results provide evidence of highly degraded triterpenes, and <sc>Spheroid echinate</sc> phytoliths are also present in sample TV003, we do not exclude the hypothesis that, apart from an indicator of palm wine or oil, the presence of these phytoliths could be an indication of the use of palm wood in pottery-making practices as well. Sample HA002 also contains traces of millets (Panicoideae), proved by the presence of <sc>Bilobate</sc>, <sc>Polylobate</sc> and <sc>Cross</sc> morphotypes. Even though millets could not be attested by the ORA on this specific sample (<italic>sensu</italic> <xref ref-type="bibr" rid="B81">Standall et al., 2022</xref>), millets were present in the city of Huelva (<xref ref-type="bibr" rid="B61">P&#x000E9;rez-Jord&#x000E0; et al., 2024</xref>) and, thus, the liquid content of the jar could have also been porridge or millet-based beverages (cf. <xref ref-type="bibr" rid="B33">Dunne et al., 2022</xref>). When compared with sample TV003 (characterized by traces of C3 cereals) and framed within an intercultural perspective emphasizing millet as typical North African crop (<xref ref-type="bibr" rid="B50">Le Moyne et al., 2023</xref>) and the city of Huelva as having a long history of Phoenician influence (<xref ref-type="bibr" rid="B60">P&#x000E9;rez-Jord&#x000E0;, 2020</xref>; <xref ref-type="bibr" rid="B61">P&#x000E9;rez-Jord&#x000E0; et al., 2024</xref>), we could advance the hypothesis that sample HA002 is indicative of a change in local agricultural practices and/or diet, which could then attest cultural exchanges between southern Spanish and North African communities.</p>
<p>Summarizing, the results of our work indicate clearly that combining phytolith analysis with ORA can be a valuable integrated method to further investigate prehistoric material culture, cultural influences, and gain information about local environmental and climatic conditions, even in the case of using post-ORA samples. Additionally, from the interpretation of our results, it is possible to assume that the phytoliths extracted and analyzed from the post-ORA pottery powder derive quite probably from both the ceramic matrix and their prolonged specific uses. The high potential of phytolith analysis to be informative even in the case of post-ORA residues can thus be attested.</p>
<p>Our results indicate, moreover, that phytoliths extraction from post organic residue analysis is not only a viable integrative method, but also that the ORA chemical pre-treatment (cf. <xref ref-type="bibr" rid="B56">Mottram et al., 1999</xref>; <xref ref-type="bibr" rid="B41">Garnier and Valamoti, 2016</xref>), being centered on the extraction of organic compounds, has not altered the content of non-organic silica material, thus the integrity of the phytoliths in our samples. We believe that the drilling process for obtaining ORA samples is unlikely to have damaged the phytoliths, as they are smaller than the resulting powder particles, and therefore should remain intact during sampling. The preservation state of the phytoliths found in our samples is excellent (cf. <xref ref-type="fig" rid="F2">Figure 2</xref>), and no broken phytoliths have emerged during the analysis at the microscope. However, a dedicated study and controlled experiments on the impact of such drilling process on phytoliths preservation should be worth pursuing. Another aspect that deserves further investigation is how variations in pottery manufacturing (e.g., kiln temperature, clay fabric, firing technology) may affect the preservation of phytoliths originally embedded in the clay, as well as how higher porosity of the vessel could result in an increased possibility of hosting a greater amount of residues in the matrix of the clay. These aspects are extremely relevant and worth being investigated further. However, they are out of the scope of this paper, since the focus of our work has been on attempting phytoliths extraction and analysis from pottery powder samples already used for Organic Residue Analysis, with the aim to see if such methodological integration performed in the same samples could provide additional information on pottery making and use, as well as of practices related to plant use, food preparation and/or consumption, cultural exchanges and networks.</p>
<p>The vivid blue compound observed in sample TR016 during the pollen extraction process is worth reflection and further investigation. As a color which rarely occurs in natural conditions, particularly in the Iberian Peninsula Early Iron Age context, the possibility of encountering this hue unintentionally is remote. Our opinion is that the chemical reaction given by the sample during the first step of the acetolysis process in the laboratory was key to determine such a specific compound; we believe that the heat during the process could have been the deciding factor in &#x0201C;awakening&#x0201D; the hue. Moreover, the lack of pollen in this sample led to the presumption of the sample being a copper-derived blue pigment. Although, based on Pliny the Elder&#x00027;s description of the production of blue pigments and dyes (<xref ref-type="bibr" rid="B67">Pliny the Elder, 1938</xref>, LCL 394: 120&#x02013;121), the use of woad (<italic>Isatis tinctoria</italic> L.) and even indigo (<italic>Indigofera tinctoria</italic>; <xref ref-type="bibr" rid="B87">Vauquelin et al., 2024</xref>) was common at the time (<xref ref-type="bibr" rid="B21">Clark et al., 1993</xref>), pollen or phytoliths from this species could be expected to be encountered in the sample. Dyeing with indigo was apparently accomplished early in European prehistory (<xref ref-type="bibr" rid="B48">Kramell et al., 2014</xref>). Woad has been the subject of several interesting studies on ancient dye techniques, currently cultivated also as a sustainable source of indigo dye (<xref ref-type="bibr" rid="B45">Hartl et al., 2015</xref>). The dye compound indigotin has been identified in many archaeological contexts, from Neolithic &#x000C7;atalh&#x000F6;y&#x000FC;k to contemporary Turkey (<xref ref-type="bibr" rid="B90">Zohary et al., 2012</xref>).</p>
<p>This particular sample, from an Early Iron Age site (fifth century BCE), was collected from El Turu&#x000F1;uelo de Guare&#x000F1;a, a ritualistic context including animal sacrifices and ceremonial burning, converting the building into a Tumulus. The sample was extracted from a coarseware fragment initially believed to be a crucible. However, this may have been the result of a glassy pigment residue on the internal surface of the fragment, a product of the heating process needed for the elaboration of copper-based pigments, particularly &#x0201C;Egyptian Blue,&#x0201D; requiring a heat between 800 &#x000B0; and 900 &#x000B0; for its production. Yet, it cannot be discarded the hypothesis that this pigment could be a lapis lazuli ultramarine. However, given the faded nature of the sample before being subjected to treatment, it is more probable that the pigment may be a copper silicate (<xref ref-type="bibr" rid="B63">Pilans, 1913</xref>). We can neither exclude the hypothesis that the vivid blue hue was the result of surface decoration of pottery materials at the final stage of production, made with vivianite (cf. <xref ref-type="bibr" rid="B29">Dillian and Bello, 2009</xref>, even though its use has not been archeologically attested in Mediterranean countries yet).</p>
<p>In conclusion, the work presented in this paper demonstrates that, without further extraction of samples on already limited material culture artifacts, post-ORA residues can be further processed for phytolith analysis, whose results&#x02014;when interpreted within an interdisciplinary framework&#x02014;can be highly informative of the complex relationships between material culture(s) and local environment(s), above all in historical contexts as prehistory, where preservation of other types of evidence may be extremely poor.</p></sec>
</body>
<back>
<sec sec-type="data-availability" id="s5">
<title>Data availability statement</title>
<p>The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: <ext-link ext-link-type="uri" xlink:href="https://doi.org/10.5281/zenodo.15371479">https://doi.org/10.5281/zenodo.15371479</ext-link>.</p>
</sec>
<sec sec-type="author-contributions" id="s6">
<title>Author contributions</title>
<p>VF: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Resources, Visualization, Writing &#x02013; original draft, Writing &#x02013; review &#x00026; editing. RA: Conceptualization, Formal analysis, Investigation, Methodology, Writing &#x02013; review &#x00026; editing. MS: Conceptualization, Formal analysis, Investigation, Methodology, Writing &#x02013; original draft, Writing &#x02013; review &#x00026; editing. EF: Conceptualization, Formal analysis, Funding acquisition, Investigation, Methodology, Resources, Visualization, Writing &#x02013; original draft, Writing &#x02013; review &#x00026; editing.</p>
</sec>
<ack><title>Acknowledgments</title><p>We would like to thank Martin Bartelheim (University of T&#x000FC;bingen), D&#x000F6;bereiner Chala-Aldana and Marta D&#x000ED;az-Zorita Bonilla (University of Granada), Clara Toscano (University of Huelva), Elena Aguilera and Raquel Zapata (Museum of Huelva), Esther Rodr&#x000ED;guez Gonz&#x000E1;lez and Sebasti&#x000E1;n Celestino P&#x000E9;rez (CSIC M&#x000E9;rida), Maxime Rageot (University of Bonn), Anneli Ekblom and Sebastian Willman (Uppsala University). We are also very thankful to the reviewers, the Chief Editor Dr. Alison Crowther and the Editor Dr. Ofir Kats, for their very relevant comments that helped improve the manuscript significantly.</p></ack>
<sec sec-type="COI-statement" id="conf1">
<title>Conflict of interest</title>
<p>The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
</sec>
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<title>Generative AI statement</title>
<p>The author(s) declared that generative AI was not 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>
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<fn-group>
<fn fn-type="custom" custom-type="edited-by" id="fn0001">
<p>Edited by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/118859/overview">Ofir Katz</ext-link>, Dead Sea and Arava Science Center, Israel</p>
</fn>
<fn fn-type="custom" custom-type="reviewed-by" id="fn0002">
<p>Reviewed by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/2089878/overview">Robert C. Power</ext-link>, University College Dublin, Ireland</p>
<p><ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/3138245/overview">Karina Chueng</ext-link>, Rio de Janeiro State University, Brazil</p>
</fn>
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<fn-group>
<fn id="fn0003"><label>1</label><p>Revert Franc&#x000E9;s, E., and Toscano, C. (in prep.). Organic Residue Analysis in Iron Age Settlements of Huelva: Insights from Pe&#x000F1;alosa, Hacienda, and Tejada la Vieja.</p></fn>
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