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<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Ecol. Evol.</journal-id>
<journal-title>Frontiers in Ecology and Evolution</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Ecol. Evol.</abbrev-journal-title>
<issn pub-type="epub">2296-701X</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.3389/fevo.2025.1605785</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Ecology and Evolution</subject>
<subj-group>
<subject>Original Research</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group>
<article-title>Trajectory of coastal wetland soil physical and microbial properties 6+ years after thin layer placement sediment amendment</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Cheng</surname>
<given-names>Jacob Z.</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2">
<sup>2</sup>
</xref>
<role content-type="https://credit.niso.org/contributor-roles/investigation/"/>
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<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Berkowitz</surname>
<given-names>Jacob F.</given-names>
</name>
<xref ref-type="aff" rid="aff3">
<sup>3</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/2896368/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
<role content-type="https://credit.niso.org/contributor-roles/supervision/"/>
<role content-type="https://credit.niso.org/contributor-roles/funding-acquisition/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>White</surname>
<given-names>John R.</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2">
<sup>2</sup>
</xref>
<xref ref-type="author-notes" rid="fn001">
<sup>*</sup>
</xref>
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<aff id="aff1">
<sup>1</sup>
<institution>Department of Oceanography &amp; Coastal Sciences, Louisiana State University</institution>, <addr-line>Baton Rouge, LA</addr-line>,&#xa0;<country>United States</country>
</aff>
<aff id="aff2">
<sup>2</sup>
<institution>Coastal Studies Institute, Louisiana State University</institution>, <addr-line>Baton Rouge, LA</addr-line>,&#xa0;<country>United States</country>
</aff>
<aff id="aff3">
<sup>3</sup>
<institution>U.S. Army Corps of Engineers, Engineer Research and Development Center</institution>, <addr-line>Vicksburg, MS</addr-line>,&#xa0;<country>United States</country>
</aff>
<author-notes>
<fn fn-type="edited-by">
<p>Edited by: Mariko Polk, North Carolina Sea Grant (NOAA), United States</p>
</fn>
<fn fn-type="edited-by">
<p>Reviewed by: Gina Marie Wimp, Georgetown University, United States</p>
<p>Nia Bartolucci, United States Environmental Protection Agency (EPA), United States</p>
<p>Ansley Levine, Texas A&amp;M University at Galveston, United States</p>
</fn>
<fn fn-type="corresp" id="fn001">
<p>*Correspondence: John R. White, <email xlink:href="mailto:jrwhite@lsu.edu">jrwhite@lsu.edu</email>
</p>
</fn>
</author-notes>
<pub-date pub-type="epub">
<day>08</day>
<month>08</month>
<year>2025</year>
</pub-date>
<pub-date pub-type="collection">
<year>2025</year>
</pub-date>
<volume>13</volume>
<elocation-id>1605785</elocation-id>
<history>
<date date-type="received">
<day>04</day>
<month>04</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>02</day>
<month>07</month>
<year>2025</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2025 Cheng, Berkowitz and White.</copyright-statement>
<copyright-year>2025</copyright-year>
<copyright-holder>Cheng, Berkowitz and White</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<p> This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). 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.</p>
</license>
</permissions>
<abstract>
<p>The intentional thin layer placement (TLP) of dredged sediment is an increasingly popular approach to maintaining marsh elevation and restoring degraded marshes, which can improve conditions for vegetation establishment. Prior TLP restoration projects, assessed shortly after construction, evaluated soil, hydrology, plant, and faunal responses. However, few long-term studies (&gt;3 yrs) investigate TLP-induced shifts in soil properties and especially properties related to biogeochemical cycling. In response, this study revisited a salt marsh 6+ years after TLP restoration and determined both soil physiochemical and microbial properties related to plant growth (nitrogen (N) mineralization) and water quality improvement (denitrification). Data were compared with samples collected before TLP project implementation and 0.5 years after project completion. Bulk density increased to 342% of the control 0.5 years after project completion and was 272% of the control after 6+ years, suggesting significant sediment retention in the marsh over time. Microbial biomass declined to 7.6% of the control following TLP, then rebounded to 29.4% of control after 6+ years. The N mineralization rate increased from 22% to 31% of control after 0.5 years and 6+ years, respectively. Notably, live root density was 3x higher in the TLP marsh compared with the control, suggesting that the restored marsh likely responded to reduced nutrient availability (approximately 1/3) by generating additional belowground biomass. TLP marsh denitrification rates were not significantly different from the control suggesting the water quality improvement ecosystem services recovers more quickly than other soil properties. While TLP soil properties appear to be trending more similar to controls over time, longer-term studies are needed to inform the ecological trajectories of sediment amended marshes.</p>
</abstract>
<kwd-group>
<kwd>wetland</kwd>
<kwd>sediment</kwd>
<kwd>marsh resilience</kwd>
<kwd>soil properties</kwd>
<kwd>restoration</kwd>
</kwd-group>
<contract-sponsor id="cn001">U.S. Army Corps of Engineers<named-content content-type="fundref-id">10.13039/100006752</named-content>
</contract-sponsor>
<counts>
<fig-count count="7"/>
<table-count count="2"/>
<equation-count count="0"/>
<ref-count count="61"/>
<page-count count="12"/>
<word-count count="5982"/>
</counts>
<custom-meta-wrap>
<custom-meta>
<meta-name>section-in-acceptance</meta-name>
<meta-value>Biogeography and Macroecology</meta-value>
</custom-meta>
</custom-meta-wrap>
</article-meta>
</front>
<body>
<sec id="s1" sec-type="intro">
<label>1</label>
<title>Introduction</title>
<p>Coastal marshes provide important ecosystem functions. Marshes have some of the highest rates of primary productivity of any ecosystem (<xref ref-type="bibr" rid="B34">Reddy and DeLaune, 2008</xref>), making them an essential component of global biogeochemical cycles. However, coastal marshes worldwide are threatened by an increasing multitude of stressors, including sea level rise (SLR), sediment starvation, subsidence, coastal edge erosion (<xref ref-type="bibr" rid="B50">Turner and Cahoon, 1987</xref>; <xref ref-type="bibr" rid="B21">DeLaune et&#xa0;al., 1978</xref>; <xref ref-type="bibr" rid="B15">Day et&#xa0;al., 2024</xref>). Sea level has been rising for over 18,000 yrs since the Holocene Era (<xref ref-type="bibr" rid="B42">Smith et&#xa0;al., 2011</xref>). However, modern rates of SLR are trending higher when compared to recent historical rates by a few mm y<sup>-1</sup> (<xref ref-type="bibr" rid="B10">Church and White, 2011</xref>). This poses challenges to communities and critical infrastructure in the coastal zone where human populations, military installations, and economic drivers (i.e., transportation hubs) are disproportionately concentrated. Generally, coastal marsh elevations keep pace with eustatic SLR by maintaining elevation via soil organic matter accretion and mineral sedimentation (<xref ref-type="bibr" rid="B18">Day et&#xa0;al., 1999</xref>). However, when SLR exceeds a marsh&#x2019;s ability to accrete material, the marsh may drown and convert to open water or mud flats (<xref ref-type="bibr" rid="B30">Orson et&#xa0;al., 1985</xref>). The degradation of marshes reduces the extent of habitat for a variety of species and increases the vulnerability of coastal communities and infrastructure to storm flooding.</p>
<p>A variety of factors influence a coastal marsh&#x2019;s ability to effectively build elevation. For example, coastal marshes receive sediment primarily through the deposition of material carried by rivers or channels from inland areas as well as from coastal waters (<xref ref-type="bibr" rid="B36">Roberts, 1997</xref>). When the major source of sediment is disconnected from the coastal marshes, the marshes experience sediment starvation, and accretion capacity is reduced (<xref ref-type="bibr" rid="B16">Day et&#xa0;al., 2007</xref>). Coastal wetlands around the globe are undergoing sediment starvation due to urban encroachment and the historical construction of dams and levees that have decreased sediment delivery by over 50% in some areas (<xref ref-type="bibr" rid="B32">Peteet et&#xa0;al., 2018</xref>; <xref ref-type="bibr" rid="B60">Yang et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B49">Torab and Azab, 2007</xref>). Subsidence consists of soil compaction and subsequent settling of the ground surface resulting in a decrease in surface elevation. Globally, subsidence can result from withdrawal of groundwater and hydrocarbons, or increased vegetation waterlogging and drowning (<xref ref-type="bibr" rid="B54">Walker et&#xa0;al., 1987</xref>). In large deltaic systems (i.e., the Mississippi River Delta), much of the subsidence is due to interstitial moisture that is forced out of the sediment structure due to the weight of overlying sediments leading to compaction (<xref ref-type="bibr" rid="B15">Day et&#xa0;al., 2024</xref>). Marsh edge erosion is driven primarily by wind and storm waves in many regions (<xref ref-type="bibr" rid="B37">Sapkota and White, 2019</xref>; <xref ref-type="bibr" rid="B39">Sapkota et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B61">Zhou et&#xa0;al., 2024</xref>). Subsidence and erosion combined with SLR increases the stress on marsh vegetation leading to open water ponding (<xref ref-type="bibr" rid="B20">DeLaune et&#xa0;al., 1994</xref>: <xref ref-type="bibr" rid="B27">Haywood et&#xa0;al., 2020</xref>). This results in a coastal landscape in critical need of rehabilitation, without which, the many coastal marsh ecosystem services delivered to communities in the region are diminished (<xref ref-type="bibr" rid="B52">Vaccare et&#xa0;al., 2019</xref>).</p>
<p>Globally, coastal wetlands offer trillions of United States dollars&#x2019; worth of ecosystem services per year (<xref ref-type="bibr" rid="B11">Costanza et&#xa0;al., 1997</xref>). Livelihoods and economies alike are increasingly threatened by wetland loss and coastal degradation (<xref ref-type="bibr" rid="B59">Williams et&#xa0;al., 1997</xref>). Resource managers are continuing to pursue restoration and rehabilitation techniques to compensate for wetland loss. As such, there are many management practices being implemented on coastlines worldwide in efforts to protect and help maintain the remaining coastal marsh. One emerging restoration management technique is the thin layer placement (TLP) of dredged sediments, which serves to nourish marsh soil with mineral material to build elevation capital and offset subsiding soils. TLP is increasingly carried out by the US Army Corps of Engineers and other entities and has been defined as the &#x201c;purposeful placement of thin layers of sediment (e.g., dredged sediment) in an environmentally acceptable manner to achieve a target elevation or thickness&#x201d;. Thin layer placement projects may include efforts to support infrastructure and/or create, maintain, enhance, or restore ecological function&#x201d; (<xref ref-type="bibr" rid="B4">Berkowitz et&#xa0;al., 2019</xref>). The TLP projects can raise marsh elevation and increase marsh resilience (<xref ref-type="bibr" rid="B33">Raposa et&#xa0;al., 2023</xref>). Thin layer placement can also improve vegetation health in previously degraded marshes via increases of elevation and bulk density, providing macrophytes with stable substrates while mitigating waterlogging (<xref ref-type="bibr" rid="B35">Reimold et&#xa0;al., 1978</xref>; <xref ref-type="bibr" rid="B57">Wilber, 1992</xref>; <xref ref-type="bibr" rid="B24">Ford et&#xa0;al., 1999</xref>; <xref ref-type="bibr" rid="B14">Davis et&#xa0;al., 2022</xref>). Coastal marshes with higher bulk density often erode more slowly, and re-establishing plants in sparsely vegetated or unvegetated areas improved soil shear strength (<xref ref-type="bibr" rid="B37">Sapkota and White, 2019</xref>). Additionally, when TLP is implemented without exceeding ecological thickness thresholds (e.g., conversion to uplands), macrofauna and infauna populations can fully recover shortly after placement (<xref ref-type="bibr" rid="B13">Croft et&#xa0;al., 2006</xref>; <xref ref-type="bibr" rid="B58">Wilber et&#xa0;al., 2007</xref>), often in &lt;3 months. TLP can also serve to restore macroinvertebrate communities (<xref ref-type="bibr" rid="B48">Tong et&#xa0;al., 2013</xref>; <xref ref-type="bibr" rid="B33">Raposa et&#xa0;al., 2023</xref>). The expediency of recovery for plant growth and fauna is one advantage of TLP which preserves seed/rhizome sources and nutrients in the rooting zone, whereas approaches such as sediment diversion and marsh creation may take years or decades to manifest suitable habitat and function.</p>
<p>Most TLP projects completed to date include very limited and essentially no long-term monitoring beyond 3 years, particularly for microbial properties. Thus, longer-term trajectories of TLP outcomes are poorly understood in the research and management community, limiting the ability of practitioners and resource managers to predict or model the environmental implications over decadal timespans. In particular, very little is known about the effects of TLP on soil microbial communities beyond very short term (&lt;1yr) monitoring periods. Microbial populations drive biogeochemical cycling, and their activity delivers wetland functions (e.g., denitrification) and associated ecosystem services (e.g., water quality improvement). Nutrient cycling and N availability, vegetation productivity, and C sequestration are all dependent on microbial-driven biogeochemical processes. Consequently, longer-term data that includes microbial properties are needed to fully understand the implications of TLP techniques for ecosystem restoration trajectories. Therefore, the goal of this study was to extend the monitoring of select soil physiochemical and microbial properties out to 6+ years at a coastal marsh TLP in Avalon, New Jersey, USA. Our approach was to compliment measurements made just before the TLP occurred and within the first year.</p>
</sec>
<sec id="s2" sec-type="materials|methods">
<label>2</label>
<title>Materials and methods</title>
<sec id="s2_1">
<label>2.1</label>
<title>Study site</title>
<p>In 2016, the US Army Corps of Engineers and project partners implemented a TLP restoration project at a rapidly degrading coastal marsh near Avalon, NJ (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). Vegetated areas of the marsh were dominated by <italic>Spartina alterniflora</italic> occupying &gt;90% absolute areal cover of the soil surface, with small amounts of <italic>Distichlis</italic> sp<italic>icata</italic> and other trace species (e.g., <italic>Salicornia Sp</italic>.) occupying &lt;5% of the soil surface marsh. The marsh soils consisted of Typic Sulfihemists mapped as the very frequently flooded Appoquinimink-Transquaking-Mispillion complex on 0 to 1 percent slopes (<xref ref-type="bibr" rid="B44">Soil Survey Staff, 2025</xref>).</p>
<fig id="f1" position="float">
<label>Figure&#xa0;1</label>
<caption>
<p>Location of the Avalon, New Jersey, USA thin-layer placement sediment project. Map inset identifies the locations of TLP locations 1 and 2 (irregular polygons), and the control marsh location (circle).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g001.tif">
<alt-text content-type="machine-generated">Map showing the northeastern United States, highlighting Philadelphia, Wilmington, and Cape May. Inset map detail displays the New Jersey Intracoastal Waterway with labeled areas: TLP 1, TLP 2, and a control.</alt-text>
</graphic>
</fig>
<p>Degradation of the back bay marsh resulted in the interspersion of vegetated areas with expanding unvegetated deepwater pools and shallow water pannes, interior marsh erosion and collapse, and other rapid marsh platform destabilization. This impaired marsh has experienced increasing inundation during which has resulted in dieback of vegetation and weaking of the soil structure via root system decay (<xref ref-type="bibr" rid="B41">Schepers et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B20">DeLaune et&#xa0;al., 1994</xref>). Following open water area expansion, vegetation can no longer re-establish for a number of reasons. First, the bulk density is reduced past the threshold required for vegetation colonization as the elevation decreases (<xref ref-type="bibr" rid="B22">DeLaune et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B17">Day et&#xa0;al., 2011</xref>). Second, water depth increased until it prevents re-establishment of vegetation. Finally, wind waves led to expansion of open water by inducing edge erosion (<xref ref-type="bibr" rid="B38">Sapkota and White, 2021</xref>). Thus, efforts were initiated to supplement the marsh elevation with dredged sediment to increase bulk density and help stabilize the marsh platform.</p>
<p>TLP was performed via high-pressure spraying of a sediment slurry (~20% sediment, 80% water) in a practice known as &#x201c;rainbowing,&#x201d; in which the dredge material is dispersed from a nozzle in an arc-shape, and by moving the dredged sediment outlet pipe into target locations surrounded by coconut fiber roll containment features (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>). There are two assessments of the Avalon, NJ TLP project published in the literature: one assessing biogeochemical properties prior to the sediment placement (<xref ref-type="bibr" rid="B5">Berkowitz et&#xa0;al., 2018</xref>) and another assessing biogeochemical properties 0.5 years after the placement (<xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al., 2018</xref>). This study extends the monitoring timeframe, now 6+ years after the TLP project initiation, to investigate the longer-term effects of TLP on wetland soil physical and microbial characteristics. In doing so, the authors of the current study compare data from all three discrete sampling intervals, evaluating the trajectory of changes in soil biogeochemical properties. The same TLP sampling stations were revisited from the two previous studies to ensure that locations were consistent across sampling temporal intervals. Sampling stations included vegetated areas that received TLP and vegetated areas that did not receive TLP to serve as unamended sediment controls (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>).</p>
<fig id="f2" position="float">
<label>Figure&#xa0;2</label>
<caption>
<p>High-pressure slurry marsh amendment technique for thin-layer placement (TLP) in the Avalon, New Jersey, USA. marsh (source: USACE).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g002.tif">
<alt-text content-type="machine-generated">A person stands on a marsh observing a large stream of dark sediment being sprayed into the air from a pipe. The marshland is interspersed with pools of water, and several people are gathered near machinery in the background.</alt-text>
</graphic>
</fig>
</sec>
<sec id="s2_2">
<label>2.2</label>
<title>Field sampling</title>
<p>In August 2023, soil samples (0&#x2013;5 cm) were collected at two vegetated TLP sites and 1 control site. Three stations were sampled at each site in duplicate using 7 cm diameter acrylic coring tubes for a total of 18 characterization samples (3 sites x 3 stations x duplicates). Samples were placed into labeled Ziploc bags and shipped overnight in a cooler on ice to the laboratory. In addition to the soil characterization samples, quadruplicate intact 20 cm long soil cores were taken from two vegetated TLP sites and one vegetated control site for a total of 12 cores for a N cycling incubation study. Cores were sealed with rubber stoppers and immediately shipped overnight on ice to the laboratory for an assessment of N mineralization and denitrification. For the original TLP, project target elevations ranged from 0.73 to 0.91m NAVD88 (<xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al., 2018</xref>).</p>
</sec>
<sec id="s2_3">
<label>2.3</label>
<title>Soil characterization</title>
<p>Soil characterization followed the same methods utilized by <xref ref-type="bibr" rid="B5">Berkowitz et&#xa0;al. (2018)</xref> and <xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al. (2018)</xref> to minimize variance between datasets. At the lab, duplicate soil samples were homogenized, and live root matter with diameter &gt;1 mm was removed. Roots were dried and weighed to determine live root mass on a dry weight basis. The 9 soil samples were analyzed for moisture content, soil dry weight bulk density, weight % organic matter, total C, N and P, extractable nutrients, extractable dissolved organic C and N, microbial biomass N (MBN), and potentially mineralizable N (PMN) rate. Briefly, moisture content was determined from weight difference of wet soil subsamples before and after placing them in a drying oven at 60&#xb0;C until constant weight (<xref ref-type="bibr" rid="B55">White and Reddy, 2000</xref>). Soil bulk density was determined by dividing the total dry weight of the sample by the volume of the core. Soil weight % organic matter (SOM) was determined by loss on ignition (LOI), which consists of ashing dried ground subsamples at 550&#xb0;C and recording the loss of mass (<xref ref-type="bibr" rid="B1">Andersen, 1976</xref>). Total P was determined via the TP ashing-acid digestion method (<xref ref-type="bibr" rid="B1">Andersen, 1976</xref>). Total C and total N were determined from dry ground subsamples analyzed on a Costech Analytical Technologies Elemental Combustion System (<xref ref-type="bibr" rid="B5">Berkowitz et&#xa0;al., 2018</xref>). Soil extractable nutrients were determined using a 2 M KCl extraction followed by filtration through a 0.45 &#x3bc;m membrane filter and acidifying samples at a pH of &lt;2 (<xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al., 2018</xref>). Samples were analyzed on a SEAL Analytical AQ300 discrete Analyzer (Mequon, WI) using USEPA methods (<xref ref-type="bibr" rid="B51">U.S. EPA, 1993</xref>). Dissolved organic C (DOC) and N (DON) were determined from a 2 M K<sub>2</sub>SO<sub>4</sub> extraction (<xref ref-type="bibr" rid="B55">White and Reddy, 2000</xref>). Microbial biomass N (MBN) was determined via chloroform fumigation-extraction method, in which non-fumigate sample N value is subtracted from fumigate samples N value to determine the amount of N contained within lysed cells (<xref ref-type="bibr" rid="B7">Brookes et&#xa0;al., 1985</xref>) with modification by <xref ref-type="bibr" rid="B55">White and Reddy (2000)</xref>. Extracted liquid samples for DOC, DON and MBN were analyzed on a Shimadzu Scientific Instrument TOC-VCSN (Columbia, MD). Potentially mineralizable nitrogen (PMN) rate was determined from the change of NH<sub>4</sub>-N production from soil under anaerobic conditions at 40&#xb0;C over a period of 10 d (<xref ref-type="bibr" rid="B55">White and Reddy, 2000</xref>). Sample were extracted @ day 0, 2, 5, and 10 of the incubation. The 2 M KCL extracted samples were analyzed on a SEAL Analytical AQ300 (<xref ref-type="bibr" rid="B51">U.S. EPA, 1993</xref>) and values were plotted over time to calculate PMN rate from linear regression (<xref ref-type="bibr" rid="B55">White and Reddy, 2000</xref>).</p>
</sec>
<sec id="s2_4">
<label>2.4</label>
<title>Water column incubation: nitrate and ammonium flux</title>
<p>Twelve intact cores, four replicates each from two different TLP areas, and four replicates from an adjacent non-TLP control area, underwent incubation to determine the ammonium flux rates and nitrate removal capacity of the wetland soil. The NH<sub>4</sub> production rate is linked to supporting plant growth and NO<sub>3</sub> reduction is linked to water quality improvement. Cores were flooded to create a 20 cm water column with salt water adjusted to a salinity of 30 to mimic site conditions using Instant Ocean<sup>&#xae;</sup>. The cores were placed into a temperature-controlled water bath at 20&#xb0;C and each core was aerated with room air to ensure aerobic conditions within the water column. The cores were spiked to produce a water column concentration of 1 mg L<sup>-1</sup> NO<sub>3</sub>-N. Seven mL water samples were collected each day for a period of 8 days, filtered through a 0.45-&#x3bc;m membrane syringe filter and acidified with H<sub>2</sub>SO<sub>4</sub> to a pH &lt;2 (<xref ref-type="bibr" rid="B9">Cheng and White, 2022</xref>). Water samples were refrigerated at 4&#xb0;C until analysis (within 2 weeks of collection) on a SEAL Analytical AQ300 colorimetric analyzer for NO<sub>3</sub>-N concentration using method 353.2 (<xref ref-type="bibr" rid="B51">U.S. EPA, 1993</xref>). Upon completion of aerobic incubation, the water column was removed and replaced, and the experiment was repeated under anaerobic water column conditions. Cores were sealed with rubber stoppers which contained inlet and outlet glass tubes and constantly bubbled with 99.99% N<sub>2</sub> gas to ensure anaerobic conditions (<xref ref-type="bibr" rid="B6">Bowes et&#xa0;al., 2022</xref>). Water samples were collected 2 times each day for a period of 3 days. Both NO<sub>3</sub>-N and NH<sub>4</sub>-N were measured using a SEAL Analytical AQ300 colorimetric analyzer, methods 353.2 and 350.1, respectively (<xref ref-type="bibr" rid="B51">U.S. EPA, 1993</xref>). Values for NO<sub>3</sub>-N and NH<sub>4</sub>-N concentrations were plotted over time on an areal basis to determine anaerobic and aerobic nitrate reduction rates and anaerobic ammonification rates, respectively.</p>
</sec>
<sec id="s2_5">
<label>2.5</label>
<title>Statistical analyses and data comparison</title>
<p>Statistics were run using R in RStudio (Version 4.3.1; Vienna, Austria). One-way ANOVA testing was used, for which a p-value of &lt;0.05 was considered significant for differences between control and TLP values as well as between time points. Shapiro-Wilk test for normality and Levene&#x2019;s test for equal variance were used to ensure that ANOVA assumptions were met. A Tukey-HSD <italic>post-hoc</italic> test was used if assumptions were met, and a Games-Howell <italic>post-hoc</italic> test was used if Tukey-HSD was violated.</p>
</sec>
</sec>
<sec id="s3" sec-type="results">
<label>3</label>
<title>Results</title>
<sec id="s3_1">
<label>3.1</label>
<title>TLP impacts on soil properties</title>
<p>The TLP-treated marsh sites had significantly higher mean bulk density than the control sites at 0.66 g cm<sup>-3</sup> and 0.24 g cm<sup>-3</sup>, respectively. The inverse was the case for gravimetric moisture content at 49.1% vs 75.8% for the TLP and control sites, respectively (<xref ref-type="table" rid="T1">
<bold>Table&#xa0;1</bold>
</xref>). These differences in bulk density and moisture content are expected, even after 6+ years, due to the addition of dense, mineral sediment and reduced soil pore space of the TLP treatment relative to the degraded, organic soil conditions of the control (<xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al., 2018</xref>). The SOM, total C, and total N were significantly lower (~30%) in the TLP soil than the control (<xref ref-type="table" rid="T1">
<bold>Table&#xa0;1</bold>
</xref>). Soil organic matter (LOI), total C, and total N were all strongly correlated (R&gt;0.9). Total P was not significantly different between TLP and control. Microbial biomass N and PMN rates in the TLP treatment soils were ~1/3 of the control marsh soils. Extractable NH<sub>4</sub>-N values were low in both the TLP and control plots, likely a result of ammonium uptake and subsequent porewater depletion during the latter part of growing season when sampling occurred (<xref ref-type="bibr" rid="B23">Feder and White, 2024</xref>). Live root mass was significantly higher (3x) in the TLP marsh compared to the control marsh (<xref ref-type="table" rid="T1">
<bold>Table&#xa0;1</bold>
</xref>). This difference is likely due to variations in soil properties and associate vegetation responses to nutrient availability, which are explored further in the discussion below.</p>
<table-wrap id="T1" position="float">
<label>Table&#xa0;1</label>
<caption>
<p>Marsh soil properties of control and thin-layer placement (TLP) treatment after 6+ years (t<sub>2</sub>).</p>
</caption>
<table frame="hsides">
<thead>
<tr>
<th valign="middle" align="left">Soil Property</th>
<th valign="middle" align="left">Units</th>
<th valign="middle" align="left">TLP</th>
<th valign="middle" align="left">Control</th>
</tr>
</thead>
<tbody>
<tr>
<td valign="middle" align="left">Extractable Ammonium</td>
<td valign="middle" align="left">mg N Kg<sup>-1</sup>
</td>
<td valign="middle" align="left">0.54 &#xb1; 0.27<sup>a</sup>
</td>
<td valign="middle" align="left">0.33 &#xb1; 0.03<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Microbial Biomass N</td>
<td valign="middle" align="left">mg N Kg<sup>-1</sup>
</td>
<td valign="middle" align="left">48.2 &#xb1; 11.8<sup>a</sup>
</td>
<td valign="middle" align="left">164 &#xb1; 68.3<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">PMN Rate</td>
<td valign="middle" align="left">mg N Kg<sup>-1</sup> d<sup>-1</sup>
</td>
<td valign="middle" align="left">15.6 &#xb1; 3.28<sup>b</sup>
</td>
<td valign="middle" align="left">50.5 &#xb1; 12.8<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Moisture Content</td>
<td valign="middle" align="left">weight %</td>
<td valign="middle" align="left">49.1 &#xb1; 3.02<sup>b</sup>
</td>
<td valign="middle" align="left">75.8 &#xb1; 1.42<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Bulk Density</td>
<td valign="middle" align="left">g cm<sup>-3</sup>
</td>
<td valign="middle" align="left">0.66 &#xb1; 0.06<sup>a</sup>
</td>
<td valign="middle" align="left">0.24 &#xb1; 0.02<sup>b</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Loss On Ignition</td>
<td valign="middle" align="left">weight %</td>
<td valign="middle" align="left">12.8 &#xb1; 1.42<sup>b</sup>
</td>
<td valign="middle" align="left">46.5 &#xb1; 3.90<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Total Carbon</td>
<td valign="middle" align="left">g C Kg<sup>-1</sup>
</td>
<td valign="middle" align="left">46.8 &#xb1; 6.22<sup>b</sup>
</td>
<td valign="middle" align="left">198 &#xb1; 9.29<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Total Nitrogen</td>
<td valign="middle" align="left">g N Kg<sup>-1</sup>
</td>
<td valign="middle" align="left">3.90 &#xb1; 0.438<sup>b</sup>
</td>
<td valign="middle" align="left">10.1 &#xb1; 0.499<sup>a</sup>
</td>
</tr>
<tr>
<td valign="middle" align="left">Total Phosphorus</td>
<td valign="middle" align="left">mg P Kg<sup>-1</sup>
</td>
<td valign="middle" align="left">801 &#xb1; 72.3<sup>a</sup>
</td>
<td valign="middle" align="left">609 &#xb1; 54.6<sup>a</sup>
</td>
</tr>
<tr>
<td valign="bottom" align="left">Live Root Mass</td>
<td valign="middle" align="left">mg cm<sup>-3</sup>
</td>
<td valign="middle" align="left">39.7 &#xb1; 9.04 <sup>a</sup>
</td>
<td valign="middle" align="left">13.1 &#xb1; 3.05 <sup>b</sup>
</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<fn>
<p>Values represent the mean &#xb1; 1 standard error (n=9). Samples collected in August 2023. Different letters between TLP and Control columns indicated significant differences at 0.05.</p>
</fn>
</table-wrap-foot>
</table-wrap>
</sec>
<sec id="s3_2">
<label>3.2</label>
<title>TLP impacts on N cycling</title>
<p>There was no significant difference between the NO<sub>3</sub>
<sup>-</sup> reduction rates of the TLP cores and the control cores under aerobic (<xref ref-type="fig" rid="f3">
<bold>Figure&#xa0;3A</bold>
</xref>) or anaerobic (<xref ref-type="fig" rid="f3">
<bold>Figure&#xa0;3B</bold>
</xref>) water column conditions. Nitrate reduction rates under aerobic water column conditions for the control averaged 45.3 &#xb1; 9.01 mg m<sup>-2</sup> d<sup>-1</sup> while mean rates for the TLP averaged 53.3 &#xb1; 4.49 mg m<sup>-2</sup> d<sup>-1</sup>. Nitrate reduction rates under anaerobic water columns were statistically higher, averaging 255 &#xb1; 27.7 mg m<sup>-2</sup> d<sup>-1</sup> for the control and 205 &#xb1; 15.8 mg m<sup>-2</sup> d<sup>-1</sup> for the TLP. Notably, the N-reduction related water quality improvement potential of these TLP marsh soils was not significantly different than the control marsh, despite the observed differences in soil properties.</p>
<fig id="f3" position="float">
<label>Figure&#xa0;3</label>
<caption>
<p>Water column incubation nitrate concentrations on an areal basis over time for both <bold>(A)</bold> aerobic and <bold>(B)</bold> anaerobic water column conditions. Data represent mean &#xb1; 1 standard error (n = 4).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g003.tif">
<alt-text content-type="machine-generated">Two line graphs showing nitrate concentration (mg N m&#xb2;) over eight days. Graph A (Aerobic) and Graph B (Anaerobic) compare Control (green), TLP 1 (blue), and TLP 2 (red) treatments. Concentrations decrease over time in both conditions, with noticeable differences in rates and final levels.</alt-text>
</graphic>
</fig>
<p>The mean anaerobic NH<sub>4</sub>
<sup>+</sup> flux rate was 44.4 &#xb1; 24.14 mg N kg<sup>&#x2212;1</sup> d<sup>&#x2212;1</sup> in both control and TLP marsh soils under anaerobic water column conditions, indicating similar N availability in both sites (<xref ref-type="fig" rid="f4">
<bold>Figure&#xa0;4</bold>
</xref>). This ammonium flux from heterotrophic soil bacterial activity, reflects the soils&#x2019; capacity to supply N to the root zone for macrophyte uptake (<xref ref-type="bibr" rid="B56">White and Reddy, 2001</xref>).</p>
<fig id="f4" position="float">
<label>Figure&#xa0;4</label>
<caption>
<p>Mean &#xb1; 1 standard error (n=4) intact core, anaerobic ammonium flux rates for two thin-layer placement (TLP) marsh sites and one control site.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g004.tif">
<alt-text content-type="machine-generated">Bar graph showing rates in milligrams per square meter per day for TLP 1, TLP 2, and Control. TLP 1 is about 40, TLP 2 slightly above 40, and Control around 50, with error bars displayed.</alt-text>
</graphic>
</fig>
<p>There was a range of changes of soil parameters and biogeochemical measures across the time points found in this study when compared with the two previous studies covering 6+ years (<xref ref-type="table" rid="T2">
<bold>Table&#xa0;2</bold>
</xref>). These differences over time are explored in more detail in the following Discussion section.</p>
<table-wrap id="T2" position="float">
<label>Table&#xa0;2</label>
<caption>
<p>Change in soil property over time for the Avalon, NJ marsh thin-layer placement (TLP) project (November 2016 (t<sub>0</sub>) (<xref ref-type="bibr" rid="B5">Berkowitz et&#xa0;al., 2018</xref>); June 2017 (t<sub>1</sub>) (<xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al., 2018</xref>); August 2023 (t<sub>2</sub>) (this study).</p>
</caption>
<table frame="hsides">
<thead>
<tr>
<th valign="top" align="center">Soil Property</th>
<th valign="top" align="center">Treatment</th>
<th valign="top" align="center">Statistical Test</th>
<th valign="bottom" align="center">F(df<sub>1</sub>, df<sub>2</sub>), p-value</th>
<th valign="top" align="center">t0 to t1 p value</th>
<th valign="top" align="center">t1 to t2 p value</th>
<th valign="top" align="center">t0 to t2 p value</th>
</tr>
</thead>
<tbody>
<tr>
<td valign="bottom" align="left">Extractable Ammonium</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,15)=5.35, p=0.018</td>
<td valign="bottom" align="center">0.893</td>
<td valign="bottom" align="center">0.087</td>
<td valign="bottom" align="center">0.009</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=2.42, p=0.131</td>
<td valign="bottom" align="center">0.53</td>
<td valign="bottom" align="center">0.429</td>
<td valign="bottom" align="center">0.114</td>
</tr>
<tr>
<td valign="bottom" align="left">Microbial Biomass N</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,15)=38.4, p&lt;0.001</td>
<td valign="bottom" align="center">0.002</td>
<td valign="bottom" align="center">0.055</td>
<td valign="bottom" align="center">0.003</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=1.63, p=0.237</td>
<td valign="bottom" align="center">0.294</td>
<td valign="bottom" align="center">0.962</td>
<td valign="bottom" align="center">0.531</td>
</tr>
<tr>
<td valign="bottom" align="left">PMN Rate</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,15)=75.0, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.073</td>
<td valign="bottom" align="center">&lt;0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=7.65, p=0.07</td>
<td valign="bottom" align="center">0.034</td>
<td valign="bottom" align="center">0.311</td>
<td valign="bottom" align="center">0.113</td>
</tr>
<tr>
<td valign="bottom" align="left">Moisture Content</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,15)=79.8, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.033</td>
<td valign="bottom" align="center">&lt;0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=3.33, p=0.0709</td>
<td valign="bottom" align="center">0.092</td>
<td valign="bottom" align="center">0.913</td>
<td valign="bottom" align="center">0.198</td>
</tr>
<tr>
<td valign="bottom" align="left">Bulk Density</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,15)=52.5, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.026</td>
<td valign="bottom" align="center">0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Games-Howell</td>
<td valign="bottom" align="left">F(2,12)=0.671, p=0.529</td>
<td valign="bottom" align="center">0.582</td>
<td valign="bottom" align="center">0.531</td>
<td valign="bottom" align="center">0.996</td>
</tr>
<tr>
<td valign="bottom" align="left">Loss On Ignition</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,15)=313.5, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.651</td>
<td valign="bottom" align="center">&lt;0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=1.58, p=0.347</td>
<td valign="bottom" align="center">0.316</td>
<td valign="bottom" align="center">0.786</td>
<td valign="bottom" align="center">0.836</td>
</tr>
<tr>
<td valign="bottom" align="left">Total Carbon</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,15)=262.8, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.221</td>
<td valign="bottom" align="center">&lt;0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=2.30, p=0.143</td>
<td valign="bottom" align="center">0.18</td>
<td valign="bottom" align="center">0.253</td>
<td valign="bottom" align="center">0.991</td>
</tr>
<tr>
<td valign="bottom" align="left">Total Nitrogen</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,15)=162.4, p&lt;0.001</td>
<td valign="bottom" align="center">&lt;0.001</td>
<td valign="bottom" align="center">0.269</td>
<td valign="bottom" align="center">&lt;0.001</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=2.127, p=0.162</td>
<td valign="bottom" align="center">0.178</td>
<td valign="bottom" align="center">0.997</td>
<td valign="bottom" align="center">0.329</td>
</tr>
<tr>
<td valign="bottom" align="left">Total Phosphorus</td>
<td valign="bottom" align="left">TLP</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,15)=1.90, p=0.185</td>
<td valign="bottom" align="center">0.464</td>
<td valign="bottom" align="center">0.76</td>
<td valign="bottom" align="center">0.165</td>
</tr>
<tr>
<td valign="bottom" align="left">
</td>
<td valign="bottom" align="left">Control</td>
<td valign="bottom" align="left">Tukey HSD</td>
<td valign="bottom" align="left">F(2,12)=11.4, p=0.002</td>
<td valign="bottom" align="center">0.365</td>
<td valign="bottom" align="center">0.001</td>
<td valign="bottom" align="center">0.01</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<fn>
<p>A p value &lt;0.05 indicates significant change between stated time points for the listed soil property within the TLP or control marsh areas.</p>
</fn>
</table-wrap-foot>
</table-wrap>
</sec>
</sec>
<sec id="s4" sec-type="discussion">
<label>4</label>
<title>Discussion</title>
<sec id="s4_1">
<label>4.1</label>
<title>TLP soil property trajectory</title>
<p>This study integrated data from the two aforementioned previous studies: <xref ref-type="bibr" rid="B5">Berkowitz et&#xa0;al. (2018)</xref>, collected in November 2016 (t<sub>0</sub>) (<xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Table S1</bold>
</xref>), and <xref ref-type="bibr" rid="B53">VanZomeren et&#xa0;al. (2018)</xref> collected 0.5 years post-TLP in June 2017 (t<sub>1</sub>; <xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Table S2</bold>
</xref>) with our data 6+ years after project implementation (t<sub>2</sub>) to analyze the trajectory of soil properties and wetland functions following project implementation. These data represent the longest investigation of soil N biogeochemical changes in TLP treated wetlands currently available in the literature that the authors are aware.</p>
<p>In November 2015 (t<sub>0</sub>), before the initiation of the project, the bulk density of the vegetated TLP marsh soil (<xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Table S1</bold>
</xref>; <xref ref-type="fig" rid="f5">
<bold>Figure&#xa0;5A</bold>
</xref>) was not significantly different from the control. However, by June 2016 (t<sub>1</sub>), approximately 0.5 years (t<sub>1</sub>) after TLP application, the bulk density in the TLP area was 342% of the control. By August 2023 (t<sub>2</sub>), the bulk density remained relatively high, at 272% of the control, which suggests long-term sediment retention related to marsh stability, important as <xref ref-type="bibr" rid="B37">Sapkota and White (2019)</xref> found that coastal marsh with higher soil bulk density eroded more slowly. The observed decrease in bulk density over time could reflect root colonization and bioturbation of the TLP sediment, along with increases in soil organic matter. Moisture content increased from 52.4% of the control at t<sub>1</sub> to 64.8% at t<sub>2</sub> (<xref ref-type="fig" rid="f5">
<bold>Figure&#xa0;5B</bold>
</xref>), showing an inverse relationship with bulk density.</p>
<fig id="f5" position="float">
<label>Figure&#xa0;5</label>
<caption>
<p>Trajectory of <bold>(A)</bold> bulk density and <bold>(B)</bold> moisture content of thin-layer placement (TLP) Marsh vs. Control sites over time. Data represent mean &#xb1; standard error (n = 6), except for the 6+ yr control time point, for which (n=3). Bold percentage represent % of control for each soil property for each time point.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g005.tif">
<alt-text content-type="machine-generated">Graph A shows bulk density change over seven years, with TLP increasing to 342% initially and decreasing to 272%, while Control remains stable at 93.9%. Graph B shows moisture content, with TLP declining from 102% to 52.4% and rising to 64.8%, while Control remains steady around 80%.</alt-text>
</graphic>
</fig>
<p>Although soil organic matter (SOM), total C, and total N as percentages of control increased on average from t<sub>1</sub> to t<sub>2</sub>, the changes were not statistically significant (<xref ref-type="fig" rid="f6">
<bold>Figures&#xa0;6A-C</bold>
</xref>; <xref ref-type="table" rid="T2">
<bold>Table&#xa0;2</bold>
</xref>). This lack of difference is also likely related to the fact that the density of organic matter remains very low compared to mineral sediment and therefore requires large changes in organic matter content to detect density differences. However, this trend is consistent with the expected gradual accumulation of organic matter in the TLP marsh. Total P increased from 84.5% of the control in t1 to 131% in t<sub>2</sub> (<xref ref-type="table" rid="T1">
<bold>Table&#xa0;1</bold>
</xref>; <xref ref-type="fig" rid="f6">
<bold>Figure&#xa0;6D</bold>
</xref>). Again, the mineral fraction will dominate total P in the sediment amended marsh soils. One recent coastal marsh study separated total P into total organic P and total inorganic P to assess the mineral-bound P. When mineral river sediment was deposited in the marsh, the overall total P did not change but the relative distribution of majority total organic P shifted to total inorganic P (<xref ref-type="bibr" rid="B45">Spera et&#xa0;al., 2020</xref>).</p>
<fig id="f6" position="float">
<label>Figure&#xa0;6</label>
<caption>
<p>Soil organic matter properties of thin-layer placement (TLP) marsh vs. control sites over time for <bold>(A)</bold> Loss on ignition (organic matter content), <bold>(B)</bold> Total soil carbon, <bold>(C)</bold> total soil nitrogen, <bold>(D)</bold> total soil phosphorus. Data represent mean &#xb1; standard error (n=6), except for the 6+ yr control time point, for which (n=3). Bold percentage represent % of control for each soil property for each time point.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g006.tif">
<alt-text content-type="machine-generated">Four graphs labeled A to D compare soil properties over seven years between Control and TLP treatments. Graph A shows loss on ignition; Graph B shows total carbon; Graph C shows total nitrogen; Graph D shows total phosphorus. Control lines are dotted, TLP are solid, with varying percentages indicating differences.</alt-text>
</graphic>
</fig>
<p>Extractable ammonium of the TLP sites (<xref ref-type="fig" rid="f7">
<bold>Figure&#xa0;7A</bold>
</xref>) did not differ significantly from the controls at any time point. Ammonium levels for all sites were notably low in t<sub>2</sub>, likely due to plant uptake. The microbial-mediated process of PMN rose from 22.9% of the control in t<sub>1</sub> to 30.9% in t<sub>2</sub> (<xref ref-type="fig" rid="f7">
<bold>Figure&#xa0;7B</bold>
</xref>), while MBN increased from 7.58% to 29.4% of the control over the same period (<xref ref-type="fig" rid="f7">
<bold>Figure&#xa0;7C</bold>
</xref>). These increases of PMN and MBN as a % of the control indicate increasing N cycling rates which are typically associated with higher organic matter soils (<xref ref-type="bibr" rid="B55">White and Reddy, 2000</xref>). While the soil trajectory of the sediment amended soils appears to be trending closer to the control, the N measures remain well below pre-project site conditions, and available data suggest that the restored wetland soil properties continue to develop along trajectories that differ from both the control and the original site conditions.</p>
<fig id="f7" position="float">
<label>Figure&#xa0;7</label>
<caption>
<p>Soil N cycling properties of thin-layer placement (TLP) Marsh vs. Control sites over time for <bold>(A)</bold> Extractable NH<sub>4</sub>-N, <bold>(B)</bold> Potentially Mineralizable N rates, <bold>(C)</bold> Microbial biomass N, and <bold>(D)</bold> Total N. Data represent mean &#xb1; standard error (n=6), except for the 6+ yr control time point, for which (n=3). Bold percentage represent % of control for each soil property for each time point.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fevo-13-1605785-g007.tif">
<alt-text content-type="machine-generated">Four graphs labeled A to D compare nitrogen metrics over seven years for TLP and Control treatments. Graph A shows extractable NH4+ decreasing for both, with TLP having higher initial and final values. Graph B presents PMN rate decline, with Control increasing by year seven. Graph C indicates a sharp decline in MBN for both, stabilizing thereafter, with Control higher at the start. Graph D shows total nitrogen rapidly declining initially, then leveling off, with TLP increasing gradually after year one. Percent changes are noted next to trend lines.</alt-text>
</graphic>
</fig>
<p>Low marsh elevation and bulk density are primary contributors to marsh degradation (<xref ref-type="bibr" rid="B17">Day et&#xa0;al., 2011</xref>). The TLP adds mineral material to the marsh soil, increasing elevation and bulk density. The restoration of these two physical factors allows for the re-establishment of vegetation and prevents further ponding and degradation. Additionally, by t<sub>1</sub>, the re-vegetation of the TLP area was already taking place with plant pushing up through the dredge material and the restored marsh was visually indistinguishable from naturally vegetated areas in the region by t<sub>2</sub> (<xref ref-type="bibr" rid="B26">Harris et&#xa0;al., 2025</xref>).</p>
<p>Examining the trajectories of the percentage difference of biogeochemical properties reveals that most soil characteristics linked to microbial activity in the TLP soil are moving towards conditions observed at control sites. However, some properties, including bulk density and organic matter content, are likely to follow distinct patterns due to the addition of mineral sediments, which instantaneously impact soil structure and composition (<xref ref-type="bibr" rid="B3">Berkowitz et&#xa0;al., 2022</xref>) (<xref ref-type="fig" rid="f5">
<bold>Figures&#xa0;5A</bold>
</xref>, <xref ref-type="fig" rid="f6">
<bold>6A</bold>
</xref>). The organic matter and total C and N each display slight, but non-significant average increases over time. In properties related to N cycling, such as total N, PMN and MBN, there is a similar, non-significant increase over time, suggesting these characteristics may ultimately converge toward control marsh levels over long time scales (<xref ref-type="fig" rid="f6">
<bold>Figures&#xa0;6A&#x2013;D</bold>
</xref>; <xref ref-type="table" rid="T2">
<bold>Table&#xa0;2</bold>
</xref>). As plants re-establish and soil organic material accumulates, further increases in organic matter and N are anticipated over time (<xref ref-type="bibr" rid="B12">Craft et&#xa0;al., 2003</xref>). Longer-term data are needed to accurately define the progression of these changes and the slope of the restoration trajectory curve. Additional data is also required to inform the application of control (or &#x2018;reference&#x2019;) conditions as milestones for determining restoration project success, or if alternative process-based measures of ecological function and service delivery yield more desirable outcomes.</p>
<p>Raising marsh elevation reduces the duration of soil inundation, leading to increased oxidation at the soil surface. In contrast, prolonged flooding creates more anaerobic conditions in surface soil (<xref ref-type="bibr" rid="B34">Reddy and DeLaune, 2008</xref>). This shift toward oxidation with higher elevation may initially accelerate the decomposition of newly deposited detrital material, potentially lowering C storage from detritus. Increased decomposition from higher elevation will affect soil properties such as PMN, MBN, TN, and TC. However, shorter inundation periods also reduce flooding stress on plants. <xref ref-type="bibr" rid="B43">Snedden et&#xa0;al. (2015)</xref> found that increased elevation led to higher plant primary proactivity and biomass, with notable growth observed at elevations up to 19 cm above mean high water&#x2014;the maximum elevation in their study. Other studies extending the soil surface elevation up to 30 and 40 cm showed that intermediate elevations best supported plant growth (<xref ref-type="bibr" rid="B28">Kirwan and Guntenspergen, 2012</xref>; <xref ref-type="bibr" rid="B29">Morris et&#xa0;al., 2013</xref>). Thus, balancing plant productivity with decomposition is crucial to optimizing ecological outcomes when implementing marsh restoration projects using TLP, where elevation gradient and target tolerances are often smaller than other coastal restoration/creation approaches.</p>
</sec>
<sec id="s4_2">
<label>4.2</label>
<title>Root density and carbon sequestration</title>
<p>As mentioned previously, the live root density in the TLP marsh soil is 3x higher than the vegetated control per unit volume of soil. One explanation for this difference is that the plants responded to a lower pool of N availability and increased root biomass to compensate. In the 2023 (t<sub>2</sub>) TLP marsh dataset, the soil contained approximately 1/3 of the TN, 1/3 of the MBN, and 1/3 of the PMN rate of the control, which are inversely proportional to the 3x increased root matter found in the TLP soils. This result suggests that the plants in the TLP marsh increased root mass in order to compensate for lower nutrient availability (<xref ref-type="bibr" rid="B19">Deegan et&#xa0;al., 2012</xref>). <xref ref-type="bibr" rid="B31">Payne et&#xa0;al. (2021)</xref> also reported higher belowground biomass in TLP marshes, approximately 50% greater root density, 3 years after the TLP project. <xref ref-type="bibr" rid="B24">Ford et&#xa0;al. (1999)</xref> reported that root biomass had reached or exceeded that of the control just 1 year after a TLP in another TLP project. Another possible explanation for the increased root growth is the absence of prior root occupation in the soil. Roots and rhizomes naturally expand in the subsurface, invading any viable nearby soil (<xref ref-type="bibr" rid="B8">Cahoon et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B24">Ford et&#xa0;al., 1999</xref>). Consequently, the unused soil space following TLP allows for the rapid expansion of root mass into previously unoccupied areas, as there is minimal belowground vegetation. This enables plant roots to quickly establish themselves in the dredged sediment, effectively filling the unclaimed &#x201c;territory&#x201d;.</p>
<p>Whether root growth is driven by physical or nutrient dynamics or a combination of factors, the increase in plant roots and rhizomes has the potential to enhance marsh sediment retention and resilience (<xref ref-type="bibr" rid="B46">Stagg and Mendelssohn, 2011</xref>). Increased root biomass strengthens soil shear resistance and reduces erosion (<xref ref-type="bibr" rid="B25">Graham and Mendelssohn, 2014</xref>; <xref ref-type="bibr" rid="B40">Sasser et&#xa0;al., 2018</xref>). The TLP is likely to contribute to coastal plant community resilience by providing new soil for roots and rhizomes to colonize. Consequently, the combined effects of increased root biomass and the greater bulk density of TLP-amended soils suggest that TLP enhances the marsh&#x2019;s capacity for storm protection and erosion resilience.</p>
<p>Thin-layer placement has the potential to be an effective method to build elevation capital and C storage strategy to enhance biogeochemical nutrient cycling. In addition to burying existing organic rich soil (<xref ref-type="bibr" rid="B2">Bartolucci and Fulweiler, 2024</xref>), TLP has been shown to stimulate increased growth of belowground biomass. Notably, primary production in marsh plants is typically higher in belowground vs aboveground biomass. <xref ref-type="bibr" rid="B47">Stagg et&#xa0;al. (2017)</xref> reported that belowground biomass in coastal marshes can be up to five times greater than aboveground biomass. Further research is needed on the longer-term biogeochemical cycling capacity and trajectory of sites restored using TLP and other techniques.</p>
</sec>
</sec>
<sec id="s5" sec-type="conclusions">
<label>5</label>
<title>Conclusions</title>
<p>The difference in soil properties, particularly the higher bulk density in the TLP, appear to have improved marsh stability and reduced edge erosion through sediment retention over time. This study could not detect any significant difference in water quality improvement (N cycling) between the TLP marsh and control areas after 6+ years after TLP project, despite the observed differences in soil physical and microbial properties. Soil physiochemical properties are trending more similar to the control marsh sites over time but are likely to remain on different trajectories depending on soil property. Further studies are needed to track these changes over longer periods of time to reduce the uncertainty in the slope of the restoration trajectory curve. The wetland function of denitrification and nutrient supply for vegetation (ammonium flux) was not statistically different between the control and TLP sites. The PMN and total N concentrations in the TLP was approximately 1/3 of the control, which may explain the threefold increase in root biomass as macrophytes seek out additional resources. After 6+ years, the restored marsh has achieved several of the functional measures or project success (e.g., denitrification rates, vegetation colonization), while some measures (e.g., SOM) continue to slowly advance toward but fail to reach control conditions. Overall, results suggest that degraded marshes can be successfully restored using TLP and that project benefits are sustained over near-decadal timescales.</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/<xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Material</bold>
</xref>. Further inquiries can be directed to the corresponding author.</p>
</sec>
<sec id="s7" sec-type="author-contributions">
<title>Author contributions</title>
<p>JC: Investigation, Writing &#x2013; original draft, Methodology. JB: Conceptualization, Methodology, Supervision, Funding acquisition, Writing &#x2013; review &amp; editing. JW: Resources, Conceptualization, Writing &#x2013; review &amp; editing, Supervision, Methodology.</p>
</sec>
<sec id="s8" sec-type="funding-information">
<title>Funding</title>
<p>The author(s) declare that financial support was received for the research and/or publication of this article. Funding was provided by the Technical Director for Military Environmental Engineering and Sciences, US Army Engineer Research and Development Center.</p>
</sec>
<ack>
<title>Acknowledgments</title>
<p>This research was developed as part of the Developing Engineering Practices for Ecosystem Design Solutions (DEEDS) project supervised by Dr. Yoko Slowey. Lee Potter is acknowledged for his assistance in field sampling and Lenore Tedesco from the Wetlands Institute acknowledged for planning and field day logistics.</p>
</ack>
<sec id="s9" 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="s10" 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>
</sec>
<sec id="s11" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
<sec id="s12" sec-type="supplementary-material">
<title>Supplementary material</title>
<p>The Supplementary Material for this article can be found online at: <ext-link ext-link-type="uri" xlink:href="https://www.frontiersin.org/articles/10.3389/fevo.2025.1605785/full#supplementary-material">https://www.frontiersin.org/articles/10.3389/fevo.2025.1605785/full#supplementary-material</ext-link>
</p>
<supplementary-material xlink:href="DataSheet1.pdf" id="SM1" mimetype="application/pdf"/>
</sec>
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