Nereis Virens Research Articles

Programmable Mechanical Properties from a Worm Jaw-Derived Biopolymer through Hierarchical Ion Exposure.

Mechanisms of biomaterial sclerotization in natural systems promise new insights into how the mechanical properties of engineered materials may be dynamically modulated. One such example involves the proteinaceous jaw of the marine sandworm, Nereis virens. Previously, the mechanical properties of the N. virens jaw were shown to be modulated by Zn binding, a property that was proposed to be enabled by the presence of the histidine-rich jaw protein, Nvjp-1. Here we demonstrate the creation of Nvjp-1-based hydrogels and show that progressive sclerotization of these hydrogels can be accomplished with hierarchical exposure to metal cations and anions. Divalent Zn cations are capable of reversibly sclerotizing the hydrogels through the formation of coordinate cross-links, an effect that is shown to be remarkably specific for Zn. Additionally, the degree of Zn-induced sclerotization is strongly influenced by the identity of the anion present in the hydrogel. Thus, the viscoelastic properties of Nvjp-1 hydrogels can be modulated through programmed, hierarchical exposure to specific cations and anions present in the sclerotizing salts. These observations have resulted in new hydrogel capabilities, such as the creation of anion-controlled shape-memory polymers, and will add to the number of control parameters that can be used to tune the properties of functional hydrogels in a dynamic manner.

Bioturbation activity of three macrofaunal species and the presence of meiofauna affect the abundance and composition of benthic bacterial communities

Given concerns of increasing rates of species extinctions, the relationship between biodiversity and ecosystem functioning has become a major research focus over the past two decades. Many studies have shown that biodiversity per se (e.g. species richness) or species-specific traits may be good predictors of changes in ecosystem function. Although numerous studies on this subject have focused on terrestrial systems, few have evaluated benthic marine systems. We used the Limecola balthica community as a model to test whether the number or identity of three well-studied macrofaunal species influence the sediment bacterial compartment, which drives important biogeochemical processes and influence ecosystem functioning. We also investigated the poorly known role of meiofauna in the interactions between macrofauna and bacteria. Eight combinations of 0–3 species were maintained in microcosms for 34 days in the presence or absence of meiofauna. The abundance and composition of the bacterial community, defined by the relative percentage of cells with a high (HNA) vs low (LNA) nucleic acid content, were measured. Species identity of macrofauna was a better indicator of changes in the microbial compartment than was species richness per se. In particular, the gallery-diffuser behaviour of the polychaete Alitta virens likely induced strong changes in sediment physical and geochemical properties with a major impact on the bacterial compartment. Moreover, the presence of meiofauna modulated the influence of macrofauna on bacterial communities. This study provides evidence that species identity provides greater explanatory power than species richness to predict changes in the bacterial compartment. We propose that multi-compartment approaches to describe interactions amongst different size classes of organisms and their ecological roles should be further developed to improve our understanding of benthic ecosystem functioning.

The influence of prey density and fish size on prey consumption in common sole (Solea solea L.)

We examined the influence of prey density and fish size on prey consumption in common sole (Solea solea L.) foraging on buried ragworm Alitta virens (Sars) (formerly known as Nereis virens (Sars)). The tested prey densities of 0.8, 2.2, 4.3 and 6.5 individuals dm−2 were exposed to common soles of either 100 g or 300 g. At each prey density common sole foraged for 48 h. At both common sole classes studied, a positive correlation between prey consumption and prey density was observed (P < 0.001). Relationships however differed between 100 and 300 g common sole. In 300 g common sole the relationship between prey consumption and prey density was linear (P < 0.001), whereas in 100 g common sole the relationship between prey density and prey eaten was polynomial (P = 0.018). Small common sole reached satiety prey consumption rates at nearly every prey density while large common sole did not reach satiation rates even at highest prey densities. The data suggest that in nature, polychaetes such as A. virens may contribute to the diet of small common sole even when they are only moderately abundant. In contrast, polychaetes may not be an ideal prey for larger common sole as indicated by the absence of satiety regardless of prey density.

Short-Term Effects of the Anti-sea Lice Therapeutant Emamectin Benzoate on Clam Worms (Nereis virens).

The polychaete Nereis virens occurs commonly in marine sediments, is widely distributed, and is a popular bait species, as well as a potential replacement for wild-caught fish in commercial fish feed preparations. It is being considered as a potential co-extractive species for culture in integrated multi-trophic aquaculture operations. However, it is not known whether pesticides or drugs used to treat sea lice on farmed salmon, such as emamectin benzoate (EB), would adversely affect cultured or wild worms, because these compounds may persist in the environment. To determine the potential effects of EB to N. virens, bioassays were performed wherein worms were exposed in sand for 30days to a concentration of 400µg/kg dw (nominal). While no treatment-related mortality occurred, significant decreases in worm mass and marked behavioral changes (lack of burrowing) were observed in EB-treated sand compared with controls. These lab-based observations suggest a potential hazard to worms at sites where EB treatments have occurred.

A matter of perception: The influence of waterborne signals from fish and conspecifics on the feeding related behavior of Alitta virens (Sars)

The present study compares the influence of signals released from fish (common sole, Solea solea L.) and conspecifics on the feeding behavior of the ragworm Alitta virens (Sars) (formerly known as Nereis virens (Sars). Experimental tanks stocked with ragworms were connected to a donor tank by a tube to allow the transfer of water born signal molecules. The donor tanks were stocked with sole and ragworms to create 4 different treatments: “control (blank)”, “ragworms”, “sole” and “sole+ragworms”. All behavioral recordings were made at 19 random days out of a total of 31 feeding days. Video recordings were made each time for 30 minutes starting when first feed entered the tank. All behavioral responses, i.e. latency, numbers and length of ragworms out of burrow were negatively affected when ragworms were exposed to water from “sole+ragworms” donor tanks. When exposed to water from “ragworms” and “sole” donor tanks, the experimental ragworms also reduced their behavioral responses but to a lesser extent than when exposed to the combination of “sole+ragworm” donor tanks. Out-of-burrow activities remained hampered in ragworms exposed to water coming from “sole” and “sole+ragworms” donor tanks at all observation days. Results indicate that signals released by damaged conspecifics and to a lesser extent fish borne cues play an important role in anti-predatory behavior of A. virens.

Interplay between flow and bioturbation enhances metal efflux from low-permeability sediments

Understanding the interplay effects between processes such as hydrodynamic forcing, sediment resuspension, and bioturbation is key to assessment of contaminated sediments. In the current study, effects of hydrodynamic forcing, sediment resuspension, and bioturbation by the marine polychaete Nereis virens were evaluated both independently and together in a six-month flume experiment. The results show that hydrodynamic forcing without resuspension or worm action slightly enhanced efflux of dissolved Cu to the water column, sediment resuspension released considerable amounts of dissolved Cu, and interactions between hydrodynamics and worm burrowing further enhanced Cu efflux. In non-bioturbated sediments, fine particles were only resuspended to the overlying water under the highest imposed shear stress, 0.58Pa. However, bioturbated sediments were resuspended under all shear stresses tested (0.11–0.58Pa), indicating that bioturbation destabilized the sediment bed. Further, increases in fluid shear following bioturbation caused rapid releases of dissolved Cu to the overlying water within a few hours. Cu efflux under fluid shears of 0.47Pa and 0.58Pa were 360× and 15× greater after the introduction of worms compared with the same flow conditions without their presence. Overall, our results indicate that the release of metals from low-permeability sediments is greatly enhanced by interactions between flow and bioturbation.

A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells

Nereis active protease (NAP) is a novel fibrinolytic active serine protease from the polychaete Nereis virens. In this study, NAP was purified from Nereis virens and the effects of NAP on human lung cancer cells were investigated. Our results indicated that NAP inhibited the proliferation and induced apoptosis of H1299 cells in a time- and dose-dependent manner. The loss of mitochondrial membrane potential, the activation of Bax and cleaved-caspase 3/9, the release of cytochrome C, and the suppression of Bcl-2 and poly-ADP ribose polymerase were observed in NAP-treated H1299 cells by flow cytometry and Western blotting. Moreover, the expression levels of Bax and Bcl-2 mRNA were determined by real-time quantitative polymerase chain reaction and the Bax/Bcl-2 expression ratio was increased in the NAP-treated cell lines. The results indicated that NAP-induced apoptosis may be related to mitochondria mediated apoptosis and occurs through caspase-dependent pathways. Then, the effects of NAP on tumor growth in animal models were observed, where 5 or 10 mg/kg of NAP noticeably reduced tumor volume and weight and increased apoptosis as determined by Western blotting when compared to the negative control group. Therefore, our findings suggest that NAP could be a hopeful anticancer medicine for its propensity to inhibit growth and induce of apoptosis in human lung cancer cells.

Palaeoecological implications of the preservation potential of soft-bodied organisms in sediment-density flows: testing turbulent waters.

Interpreting how far organisms within fossil assemblages may have been transported and if they all originated from the same location is fundamental to understanding whether they represent true palaeocommunities. In a three-factorial experimental design, we used an annular flume to generate actualistic sandy sediment-density flows that were fast (2 ms−1) and fully turbulent in order to test the effects of flow duration, sediment concentration, and grain angularity on the states of bodily damage experienced by the freshly euthanized polychaete Alitta virens. Results identified statistically significant effects of flow duration and grain angularity. Increasing sediment concentration had a statistically significant effect with angular sediment but not with rounded sediment. Our experiments demonstrate that if soft-bodied organisms such as polychaetes were alive and then killed by a flow then they would have been capable of enduring prolonged transport in fast and turbulent flows with little damage. Dependent upon sediment concentration and grain angularity, specimens were capable of remaining intact over flow durations of between 5 and 180 min, equating to transport distances up to 21.6 km. This result has significant palaeoecological implications for fossil lagerstätten preserved in deposits of sediment-density flows because the organisms present may have been transported over substantial distances and therefore may not represent true palaeocommunities.

Regeneration of the nervous and muscular system after caudal amputation in the polychaete Alitta virens (Annelida: Nereididae)

Regenerating segments in polychaetes offer a vivid example of epimorphic recovery of the lost organs and tissues. It is also a promising object for studying positional information and the mechanisms maintaining the body integrity. With the aim to develop a convenient standardized model, we described the dynamics of recovery of the major anatomical structures and created a staging system for the caudal regeneration in Alitta virens. In average the normal organization of the posterior body end is restored within 10 days after amputation (dpa). The whole regenerative process was divided into 5 stages: (1) wound healing (0–1 dpa), (2) blastema formation (1–2 dpa), (3) patterning and growth of the blastema (2–3 dpa), (4) differentiation of the first regenerated segment (3–5 dpa), (5) formation and differentiation of the subsequent 5–6 segments (5–10 dpa). The regeneration is carried out mainly by epimorphosis, although the elements of intercalary growth as well as the morphallactic transformation of the stump have been noted. Terminal structures of the pygidium (muscles of the anal sphincter, pygidial cavity, pygidial ring nerve, pygidial cirri) appear at stages 1–3, and then (from stage 3) the formation of new metameres begins in front of the pygidium. Differentiation of the first newborn segment is associated with the tissue remodeling in the last old segment. Formation of the next segments resembles accelerated postlarval growth. The neural elements of the regenerative bud are developing faster than the surrounding muscles. The neurites extending from the CNS and PNS come to the surface of the wound epithelium at stage 1. Later, nerve fibers from the CNS lengthen and thicken along with the growth of the regenerative bud. Ganglion, parapodial nerves, oblique muscles and coeloms of the first segment are detected at stage 4. Longitudinal muscles regenerate in anterior to posterior progression, being constantly in contact with the corresponding fibers of the old tissues. All other muscles differentiate from blastemal cells in isolation from the old musculature of the stump. Our data promote the further using of the posterior body end regeneration in A. virens as an experimental model for resolving crucial problems of developmental biology.

Effect of retinoids on Post2 Нох gene expression in nereid polychaetes

Retinoic acid (RA) plays an important role in vertebrate development and regeneration. RA signalling directly regulates the expression of Hox genes, being in this way involved in the patterning of the anterior-posterior (AP) axis of vertebrate embryos. So far the relationship between retinoic acid signalling and Hox genes has been shown only for chordates. In this study we incubated juvenile worms and regenerating worms of two polychaete species from the family Nereididae, Alitta virens and Platynereis dumerilii, with all-trans-retinal, the precursor of retinoic acid. Under the influence of all-trans-retinal the anterior expression boundary of Post2 Нох gene shifted towards the anterior end both in intact and in regenerating worms of both species. Our data indicate the existence of a relationship between RA signalling and Нох genes in Protostomia.

Peculiarities of isolated blastomere development of the polyhaete Alitta virens

The ability of regulation, including restoration of the missing parts, to normalize the development, is a fundamental characteristic of living systems. However, until recently, representatives of the group Spiralia (animals with spiralian development) have been considered a typical example of determinative development. Meanwhile, studies in recent years give reason to speak about the presence of inductive cell-cell interactions at the early stages of Spiralian embryo development. In this paper, for the first time, we have shown the development of the polychaete Alitta virens blastomeres isolated at different stages. The results indicate the inability of A. virens partial embryos of regulation to form a whole normal larva and a high degree of autonomous determination of trochoblast cells. At the same time, the findings support probable early induction of blastomere D and acquisition of competence by blastomere D to further development of trochoblast ciliated cells.

A comparison between marine and terrestrial invertebrate meals for mirror carp (Cyprinus carpio ) diets: Impact on growth, haematology and health

Invertebrate meals (e.g. polychaetes and insects) present novel and sustainable high-quality nutrient sources for use in fish feed formulations. To test this innovative source, an eleven-week feeding trial was conducted evaluating the effects of replacing the fishmeal (FM) component as an example of a superior protein source (FM CTRL) with ragworm meal (RW, Nereis virens) and/or silkworm pupae (SWP, Bombyx mori) in mirror carp (Cyprinus carpio) diets. Three experimental diets with partial replacement of FM (diets: RW + FM, SWP + FM and RW + SWP + FM) were formulated. All diets were formulated to be iso-nitrogenous, iso-lipidic and iso-energetic. Growth performance and feed utilization indices were assessed, and the feeding trial concluded with the analysis of haematological parameters to provide an indication of carp physiological and health status. Mean weight gain was greatest in mirror carp fed RW + FM (60.83 fish−1 day−1; P < 0.05 vs. all other diets) followed by SWP + FM (40.62 g fish−1 day−1; P < 0.05 vs. all other diets). The least weight gain was achieved in fish fed FM + SWP + RW+ and FM CTRL (34.34 and 33.96 g fish−1 day−1, respectively; not significantly different from each other). Fish fed on RW + FM diet had significantly lower plasma ammonia concentrations than any other dietary groups (P = 0.04). Mirror carp fed on SWP + FM diet (111.52 units mL−1) were observed to have a marked enhancement in alternative complement activity than FM CTRL (79.21 units mL−1, P = 0.041). Both ragworm and silkworm pupae meal present attractive sustainable functional feed component in carp diets, with benefits on enhancing growth performance and specific physiological parameters.

Ion Effect and Metal-Coordinated Cross-Linking for Multiscale Design of Nereis Jaw Inspired Mechanomutable Materials.

The Nvjp-1 protein is a key component in the jaws of Nereis virens, a species of marine worm. It contains over 25 mol % of histidine, which is believed to play a key role in the metal-coordinated cross-linking responsible for the structural stability and exceptional mechanical performance of the worm jaw. Understanding the nanoscale mechanism behind this cross-linking and its pathway in affecting the macroscopic mechanical behavior of the material is crucial to develop bioinspired mechanomutable materials based on Nvjp-1. Here, we use a combination of multiscale modeling and experimental synthesis to understand the behavior of this heterologous-expressed protein from the nano- to the macroscale. We have built a bottom-up molecular-based model, which includes electronic-based density functional theory calculations, atomistic simulation of the nanoscale properties with replica exchange molecular dynamics, and an elastic network model for describing the macroscale behavior at different pHs. This multiscale modeling supports the experimental synthesis of a photo-cross-linked Nvjp-1 hydrogel by proving both the nanoscale mechanisms and mechanical behavior predictions. Our theoretical results agree well with the experimental observations, showing that Nvjp-1 forms a more compact structure in the presence of Zn2+ ions with a suitable pH environment, leading to the formation of more stable intramolecular metal-coordinated cross-links. These metal-coordinated cross-links induce nanoscale aggregation of Nvjp-1, which is responsible for the hydrogel contraction observed in experiments and predicted by the model.

Bait worms: a valuable and important fishery with implications for fisheries and conservation management

AbstractBait is an integral part of coastal life, but is perceived as a low‐value resource as fisheries are data‐limited, locally focussed and largely unregulated even though the ecological impacts of collection are considerable. An empirical assessment of three UK‐based ragworm fisheries combined with an analysis of published literature has produced the first global assessment of polychaete bait fisheries. The five most expensive (retail price per kg) marine species sold on the global fisheries market are polychaetes (Glycera dibranchiata, Diopatra aciculata, Nereis (Alitta) virens, Arenicola defodiens and Marphysa sanguinea). We estimate that 1600 t of N. virens per annum (worth £52 million) are landed in the UK with approximately 121 000 tonnes of polychaetes collected globally valued at £5.9 billion. Using remote closed circuit television (CCTV) cameras to monitor collectors, activity at local sites is considerable with a mean of 3.14 collectors per tide (day and night) at one site and individuals digging for up to 3 h per tide, although intensity differed seasonally and between sites. Collectors removed on average 1.4 kg of N. virens per person per hour, walking a considerable distance across the intertidal sediment to reach areas that were usually already dug. The implications of these human activity and biomass removal levels are explored in the context of fisheries and conservation management. At local, regional and national scales, polychaete bait fisheries are highly valuable, extract significant biomass and have considerable impacts; therefore, they urgently require governance equivalent to other fisheries.

Early mesodermal expression of Hox genes in the polychaete Alitta virens (Annelida, Lophotrochozoa).

Hox genes are the key regulators of axial regionalization of bilaterian animals. However, their main function is fulfilled differently in the development of animals from different evolutionary branches. Early patterning of the developing embryos by Hox gene expression in the representatives of protostomes (arthropods, mollusks) starts in the ectodermal cells. On the contrary, the instructive role of the mesoderm in the axial patterning was demonstrated for vertebrates. This makes it difficult to understand if during the axial regionalization of ancestral bilaterians Hox genes first expressed in the developing mesoderm or the ectoderm. To resolve this question, it is necessary to expand the number of models for investigation of the early axial patterning. Here, we show that three Hox genes of the polychaete Alitta virens (formerly Nereis virens, Annelida, Lophotrochozoa)-Hox2, Hox4, and Lox5-are expressed in the mesodermal anlagen of the three future larval chaetigerous segments in spatially colinear manner before the initiation of Hox expression in the larval ectoderm. This is the first evidence of sequential Hox gene expression in the mesoderm of protostomes to date.

Mesoderm patterning and morphogenesis in the polychaete Alitta virens (Spiralia, Annelida): Expression of mesodermal markers Twist, Mox, Evx and functional role for MAP kinase signaling

Mesoderm represents the evolutionary youngest germ layer and forms numerous novel tissues in bilaterian animals. Despite the established conservation of the gene regulatory networks that drive mesoderm differentiation (e.g. myogenesis), mechanisms of mesoderm specification are highly variable in distant model species. Thus, broader phylogenetic sampling is required to reveal common features of mesoderm formation across bilaterians. Here we focus on a representative of Spiralia, the marine annelid Alitta virens, whose mesoderm development is still poorly investigated on the molecular level. We characterize three novel early mesodermal markers for A. virens – Twist, Mox, and Evx – which are differentially expressed within the mesodermal lineages. The Twist mRNA is ubiquitously distributed in the fertilized egg and exhibits specific expression in endomesodermal- and ectomesodermal-founder cells at gastrulation. Twist is expressed around the blastopore and later in a segmental metameric pattern. We consider this expression to be ancestral, and in support of the enterocoelic hypothesis of mesoderm evolution. We also revealed an early pattern of the MAPK activation in A. virens that is different from the previously reported pattern in spiralians. Inhibition of the MAPK pathway by U0126 disrupts the metameric Twist and Mox expression, indicating an early requirement of the MAPK cascade for proper morphogenesis of endomesodermal tissues.

Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata.

Venom peptides found in terebrid snails expand the toolbox of active compounds that can be applied to investigate cellular physiology and can be further developed as future therapeutics. However, unlike other predatory organisms, such as snakes, terebrids produce very small quantities of venom, making it difficult to obtain sufficient amounts for biochemical characterization. Here, we describe the first recombinant expression and characterization of terebrid peptide, teretoxin Tgu6.1, from Terebra guttata. Tgu6.1 is a novel forty-four amino acid teretoxin peptide with a VI/VII cysteine framework (C–C–CC–C–C) similar to O, M and I conotoxin superfamilies. A ligation-independent cloning strategy with an ompT protease deficient strain of E. coli was employed to recombinantly produce Tgu6.1. Thioredoxin was introduced in the plasmid to combat disulfide folding and solubility issues. Specifically Histidine-6 tag and Ni-NTA affinity chromatography were applied as a purification method, and enterokinase was used as a specific cleavage protease to effectively produce high yields of folded Tgu6.1 without extra residues to the primary sequence. The recombinantly-expressed Tgu6.1 peptide was bioactive, displaying a paralytic effect when injected into a Nereis virens polychaete bioassay. The recombinant strategy described to express Tgu6.1 can be applied to produce high yields of other disulfide-rich peptides.

Diet of an underappreciated benthic intertidal fish, &lt;em&gt;Cryptacanthodes maculatus&lt;/em&gt; (Cryptacanthodidae), in eastern Maine, USA

We discovered through gut-content analysis over a 20-month period (June 2009–March 2011) that a little-known benthic fish, the wrymouth, Cryptacanthodes maculatus, preyed upon soft-bottom fauna from 8 phyla, 10 classes, and 28 genera in the lower intertidal/ shallow subtidal zone in Cobscook Bay (eastern Maine, USA). Wrymouth (N = 42; total length = 30–50 cm) live infaunally in extensive subterranean burrows, and consumed mainly polychaetes and amphipods that occurred in 80% and 75% of stomachs, respectively. The two most common polychaetes, Alitta virens and Harmothoe imbricata, were found in 60% and 25% of stomachs, respectively. Gammarideans Leptocheirus pinguis and Ampelisca abdita were the two most common amphipods occurring in 40% and 33% of stomachs, respectively. Benthic core samples taken from the mid- and low shore were ambiguous in helping to understand where wrymouth forage. Wrymouth likely contribute to trophic linkages between the intertidal and subtidal zones in soft-bottoms; however, its relative role in regulating benthic community structure, aspects of its reproductive behavior, and early life-history is unknown due in part to its cryptic lifestyle. In addition to identifying components of the wrymouth’s diet, we document gravid females for the first time, and when they appear in the population.

Modeling of Bioaccumulation in Marine Benthic Invertebrates Using a Multispecies Experimental Approach.

The causal links between species traits and bioaccumulation by marine invertebrates are poorly understood. We assessed these links by measuring and modeling polychlorinated biphenyl bioaccumulation by four marine benthic species. Uniformity of exposure was achieved by testing each species in the same aquarium, separated by enclosures, to ensure that the observed variability in bioaccumulation was due to species traits. The relative importance of chemical uptake from pore water or food (organic matter, OM) ingestion was manipulated by using artificial sediment with different OM contents. Biota sediment accumulation factors (BSAFs) ranged from 5 to 318, in the order Nereis virens < Arenicola marina ≈ Macoma balthica < Corophium volutator. Calibration of a kinetic model provided species-specific parameters that represented the key species traits, thus illustrating how models provide an opportunity to read across benthic species with different feeding strategies. Key traits included species-specific differentiation between (1) ingestion rates, (2) ingestion of suspended and settled OM, and (3) elimination rates. The high BSAF values and their concomitant variability across the species challenges approaches for exposure assessment based on pore water concentration analysis and equilibrium partition theory. We propose that combining multienclosure testing and modeling will substantially improve exposure assessment in sediment toxicity tests.

The effect of sediment mixing on mercury dynamics in two intertidal mudflats at Great Bay Estuary, New Hampshire, USA

Estuarine sediments store particulate contaminants including mercury (Hg). We studied Hg sediment dynamics in two intertidal mudflats at Great Bay estuary, NH, over multiple years. Sediments at both mudflats were physically mixed down to ~10cm, as determined by 7Be measurements, albeit via different mechanisms. Portsmouth mudflat (PT) sediments were subject to bioturbation by infaunal organisms and Squamscott mudflat (SQ) sediments were subject to erosion and redeposition. The presence of higher concentrations of fresh Fe(III) hydroxide at PT suggested bioirrigation by the polychaetes (Nereis virens). At depths where infaunal bioirrigation was observed, pore-water inorganic Hg (Hgi) and methylmercury (MeHg) were lower potentially due to their interaction with Fe(III) hydroxide. Methylmercury concentrations increased immediately below this zone in some samples, suggesting that the observed increase in material flux in bioirrigated sediments may initiate from lower depths. Pore water in sediment at PT also had higher fractions of more protein-like and labile DOC than those at SQ that can lead to increased MeHg production in PT, especially at depths where Hgi is not removed from solution by Fe(III) hydroxide. Where sediment erosion and redeposition were observed at SQ, Hg species distribution was extended deeper into the sediment column. Moreover, methyl coenzyme M reductase (MCR) and mercury reductase (mer-A) genes were higher at SQ than PT suggesting differences in conditions for Hg cycling. Results showed that the near-surface region of high MeHg concentrations commonly observed in unmixed sediments does not exist in physically mixed sediments that are common in many estuarine environments.