Locomotory Palp Function in Interstitial Annelids.

AbstractThe interstitial environment of marine sediments is a complex network of voids and pores that is inhabited by a diverse and abundant fauna. Animals living within these interstitial spaces show widespread functional adaptations to this environment and have developed many strategies for moving and navigating through small spaces. Interstitial annelids demonstrate a remarkable level of morphologic diversity, and some possess dexterous, filiform palps (tentacle-like appendages common across Annelida). The function(s) of these palps in interstitial spaces has not been closely examined, and we propose that they serve a sensory role in the navigation of interstitial spaces. We investigated the locomotory function of long, dexterous palps in three families of interstitial annelids to determine their role in interstitial navigation. We observed two species of protodrilids (Protodrilidae), Pharyngocirrus eroticus (Saccocirridae), and Protodorvillea recuperata (Dorvilleidae), as they moved through two transparent sand analogs: cyolite and glass beads. All four species of annelids consistently used their palps to probe the interstitial environment while locomoting, and the distance probed with their palps was greater than the distance traveled with their heads, indicating a sensory form of palp-based navigation. The functionality of palps as sensory organs in the interstitial environment raises interesting questions about interstitial navigation and how fauna without appendages map their surroundings. The discovery of this previously undocumented function was possible only through the direct observation of interstitial behavior and emphasizes the importance of developing new techniques to study these animals in more natural habitats.

Fundamentals of burrowing in soft animals and robots.

Creating burrows through natural soils and sediments is a problem that evolution has solved numerous times, yet burrowing locomotion is challenging for biomimetic robots. As for every type of locomotion, forward thrust must overcome resistance forces. In burrowing, these forces will depend on the sediment mechanical properties that can vary with grain size and packing density, water saturation, organic matter and depth. The burrower typically cannot change these environmental properties, but can employ common strategies to move through a range of sediments. Here we propose four challenges for burrowers to solve. First, the burrower has to create space in a solid substrate, overcoming resistance by e.g., excavation, fracture, compression, or fluidization. Second, the burrower needs to locomote into the confined space. A compliant body helps fit into the possibly irregular space, but reaching the new space requires non-rigid kinematics such as longitudinal extension through peristalsis, unbending, or eversion. Third, to generate the required thrust to overcome resistance, the burrower needs to anchor within the burrow. Anchoring can be achieved through anisotropic friction or radial expansion, or both. Fourth, the burrower must sense and navigate to adapt the burrow shape to avoid or access different parts of the environment. Our hope is that by breaking the complexity of burrowing into these component challenges, engineers will be better able to learn from biology, since animal performance tends to exceed that of their robotic counterparts. Since body size strongly affects space creation, scaling may be a limiting factor for burrowing robotics, which are typically built at larger scales. Small robots are becoming increasingly feasible, and larger robots with non-biologically-inspired anteriors (or that traverse pre-existing tunnels) can benefit from a deeper understanding of the breadth of biological solutions in current literature and to be explored by continued research.

Impacts of infauna, worm tubes, and shell hash on sediment acoustic variability and deviation from the viscous grain shearing model.

Infauna influence geoacoustic parameters in surficial marine sediments. To investigate these effects, an experiment was conducted in natural sand-silt sediment in the northern Gulf of Mexico. In situ acoustic measurements of sediment sound speed, attenuation, and shear speed were performed, and sediment cores were collected from the upper 20 cm of the seabed. Laboratory measurements of sound speed and attenuation in the cores were conducted, after which the core contents were analyzed for biological and physical properties. Since no model currently accounts for the effects of infauna, a deviation from model predictions is expected. To assess the extent of this, acoustic measurements were compared with the viscous grain shearing model from Buckingham [J. Acoust. Soc. Am. 122, 1486 (2007); J. Acoust. Soc. Am. 148, 962 (2020)], for which depth-dependent profiles of sediment porosity and mean grain size measured from the cores were used as input parameters. Comparison of acoustic results with distributions of infauna, worm tubes, and shell hash suggests biogenic impacts on acoustic variability and model accuracy are important in surficial marine sediments. The presence of infauna and worm tubes were correlated with higher variability in both sound speed and attenuation and greater deviation from the model near the sediment-water interface.

Promoting a More Integrated Approach to Structure and Function.

The connection between structure and function is one of the fundamental tenets of biology: a biological unit's structure determines its function, and, conversely, its function depends upon its structure. Historically, important advances have been made either when understanding of structure leads to questions about function or when understanding of function raises questions about the structures involved. Consequently, considering the connections between structure and function from a broader perspective might lead to the development of novel hypotheses that move our understanding of the fundamental connections between structure and function forward. Better integration of structure and function is a key component in the broader goal of reintegrating biology within and across scales. Here, we provide examples of how integrating studies of structure and function as well as comparing structure-function relationships across biological scales can lead to scientific advances. We also emphasize the potential of integrating studies of structure and function across scales for bio-inspired design and for improving biology education.

Transport of biodeposits and benthic footprint around an oyster farm, Damariscotta Estuary, Maine.

The benthic impact of aquaculture waste depends on the area and extent of waste accumulation on the sediment surface below and around the farm. In this study we investigated the effect of flow on biodeposit transport and initial deposition by calculating a rough aquaculture "footprint" around an oyster aquaculture farm in the Damariscotta River, ME. We also compared a site under the farm to a downstream "away" site calculated to be within the footprint of the farm. We found similar sediment biogeochemical fluxes, geochemical properties and macrofaunal communities at the site under the farm and the away site, as well as low organic enrichment at both sites, indicating that biodeposition in this environment likely does not have a major influence on the benthos. To predict accumulation of biodeposits, we measured sediment erodibility under a range of shear stresses and found slightly higher erosion rates at the farm than at the away site. A microalgal mat was observed at the sediment surface in many sediment cores. Partial failure of the microalgal mat was observed at high shear velocity, suggesting that the mat may fail and surface sediment erode at shear velocities comparable to or greater than those calculated fromin situ flow measurements. However, this study took place during neap tide, and it is likely that peak bottom velocities during spring tides are high enough to periodically "clear" under-farm sediment of recent deposits.

Dynamics of Mud Blister Worm Infestation and Shell Repair by Oysters.

AbstractMud blister worms bore into oyster shells; and oysters respond to shell penetration by secreting new layers of shell, resulting in mud blisters on inner surfaces of oyster shells. We conducted two experiments in off-bottom oyster farms along Alabama's coast in summer 2017 to explore the dynamics of worm infestation, blister formation, and shell repair. Results support our hypothesis that only a small proportion of worms that bore into oysters cause blisters. Triploid oysters had fewer blisters than diploids, likely because of faster growth and shell repair. We treated oysters to remove mud blister worms, redeployed them at intertidal and subtidal sites for nine weeks, and found that reinfestation by worms occurred only in subtidal oysters. Intertidally deployed oysters showed no visible blister coverage, indicating recovery, whereas blister coverage increased in subtidal oysters. Reinfestation of subtidal oysters was correlated with previous burrow damage, visualized with X-ray images, thus supporting our hypothesis that worms preferentially settle in previously infested shells. Forces required to break blisters, measured with a custom-built shucking knife with an integrated force sensor, were low relative to forces required to shuck oysters, possibly because our experiment was conducted when worm infestation was increasing. Higher forces were required to break smaller, lighter-colored blisters, consistent with blister recovery; but results were highly variable and not consistent across sites and sampling times, suggesting that size and color of blisters alone did not explain shell strength. Our results indicate that oysters repair shells slowly relative to more dynamic patterns of worm infestation.

Impacts of simulated infaunal activities on acoustic wave propagation in marine sediments.

The activities of infaunal organisms, including feeding, locomotion, and home building, alter sediment physical properties including grain size and sorting, porosity, bulk density, permeability, packing, tortuosity, and consolidation behavior. These activities are also known to affect the acoustic properties of marine sediments, although previous studies have demonstrated complicated relationships between infaunal activities and geoacoustic properties. To avoid difficulties associated with real animals, whose exact locations and activities are unknown, this work uses artificial burrows and simulates infaunal activities such as irrigation, compaction, and tube building in controlled laboratory experiments. The results show statistically significant changes in sound speed and attenuation over a frequency range of 100-400 kHz, corresponding to wavelengths on the order of the burrow diameter. The greatest effects were observed for tubes constructed of hard shells which increased the attenuation by ∼30 dB m-1 across the measurement band. These results highlight the importance of biogenic hard structures such as tubes on sound attenuation and suggest that organisms that create hard structures may be good targets for acoustic mapping of infaunal abundance and distribution.

Kinematics of burrowing by peristalsis in granular sands.

Peristaltic burrowing in muds applies normal forces to burrow walls, which extend by fracture, but the kinematics and mechanics of peristaltic burrowing in sands has not been explored. The opheliid polychaete Thoracophelia mucronata uses direct peristalsis to burrow in beach sands, with kinematics consistent with the 'dual anchor system' of burrowing described for diverse organisms. In addition to expansions associated with a constrictive direct peristaltic wave, worms alternately expand the head region, which is separated by septa from the open body cavity, and expansible lateral ridges that protrude from the 10th setiger. Tracking of chaetae with fluorescent dye showed that the body wall advances while segments are thin, then stationary segments expand, applying normal forces to burrow walls. These normal forces likely compact burrow walls and serve as anchors. Perhaps more importantly, peristaltic movements minimize friction with the burrow wall, which would expand dilatant sands. Considerable slipping of worms burrowing in a lower-density sand analog suggests that this dual-anchor peristaltic burrowing may be limited to a narrow range of mechanical properties of substrata, consistent with the limited habitat of T. mucronata in a narrow swash zone on dissipative beaches.

Functional Morphology of Eunicidan (Polychaeta) Jaws.

Polychaetes exhibit diverse feeding strategies and diets, with some species possessing hardened teeth or jaws of varying complexity. Species in the order Eunicida have complex, rigid, articulated jaws consisting of multiple pairs of maxillae and a pair of mandibles. While all Eunicida possess this general jaw structure, several characteristics of the jaws vary considerably among families. These differences, described for fossilized and extant species' jaws, have been used to infer evolutionary relationships. Little has been done, however, to relate jaw functional morphology and feeding behavior to diet. To explore these relationships, we compared the jaw kinematics and morphology of two distantly related eunicidan taxa with superficially similar jaw structures: Diopatra spp. (Onuphidae), predominantly herbivorous and tube dwelling, and Lumbrineris spp. (Lumbrineridae), a burrowing carnivore. Jaw kinematics were observed by filming individuals biting in a number of orientations. Some differences in jaw structure and kinematics between Diopatra spp. and Lumbrineris spp. can be interpreted to be consistent with their differences in diet. Relating jaw morphology to diet would improve understanding of early annelid communities by linking fossil teeth (scolecodonts) to the ecological roles of extant species with similar morphologies.

Burrowing by small polychaetes - mechanics, behavior and muscle structure of Capitella sp.

Worms of different sizes extend burrows through muddy sediments by fracture, applying dorso-ventral forces that are amplified at the crack tip. Smaller worms displace sediments less than larger worms and therefore are limited in how much force they can apply to burrow walls. We hypothesized that small worms would exhibit a transition in burrowing mechanics, specifically a lower limit in body size for the ability to burrow by fracture, corresponding with an ontogenetic transition in muscle morphology. Kinematics of burrowing in a mud analog, external morphology and muscle arrangement were examined in juveniles and adults of the small polychaete Capitella sp. We found that it moves by peristalsis, and no obvious differences were observed among worms of different sizes; even very small juveniles were able to burrow through a clear mud analog by fracture. Interestingly, we found that in addition to longitudinal and circular muscles needed for peristaltic movements, left- and right-handed helical muscles wrap around the thorax of worms of all sizes. We suggest that in small worms helical muscles may function to supplement forces generated by longitudinal muscles and to maintain hydrostatic pressure, enabling higher forces to be exerted on the crack wall. Further research is needed, however, to determine whether surficial sediments inhabited by small worms fail by fracture or plastically deform under forces of the magnitudes applied by Capitella sp.

The biomechanics of burrowing and boring.

Burrowers and borers are ecosystem engineers that alter their physical environments through bioturbation, bioirrigation and bioerosion. The mechanisms of moving through solid substrata by burrowing or boring depend on the mechanical properties of the medium and the size and morphology of the organism. For burrowing animals, mud differs mechanically from sand; in mud, sediment grains are suspended in an organic matrix that fails by fracture. Macrofauna extend burrows through this elastic mud by fracture. Sand is granular and non-cohesive, enabling grains to more easily move relative to each other, and macrofaunal burrowers use fluidization or plastic rearrangement of grains. In both sand and mud, peristaltic movements apply normal forces and reduce shear. Excavation and localized grain compaction are mechanisms that plastically deform sediments and are effective in both mud and sand, with bulk excavation being used by larger organisms and localized compaction by smaller organisms. Mechanical boring of hard substrata is an extreme form of excavation in which no compaction of burrow walls occurs and grains are abraded with rigid, hard structures. Chemical boring involves secretion to dissolve or soften generally carbonate substrata. Despite substantial differences in the mechanics of the media, similar burrowing behaviors are effective in mud and sand.

Diet of worms emended: an update of polychaete feeding guilds.

Polychaetes are common in most marine habitats and dominate many infaunal communities. Functional guild classification based on taxonomic identity and morphology has linked community structure to ecological function. The functional guilds now include osmotrophic siboglinids as well as sipunculans, echiurans, and myzostomes, which molecular genetic analyses have placed within Annelida. Advances in understanding of encounter mechanisms explicitly relate motility to feeding mode. New analyses of burrowing mechanics explain the prevalence of bilateral symmetry and blur the boundary between surface and subsurface feeding. The dichotomy between microphagous deposit and suspension feeders and macrophagous carnivores, herbivores, and omnivores is further supported by divergent digestive strategies. Deposit feeding appears to be limited largely to worms longer than 1 cm, with juveniles and small worms in general restricted to ingesting highly digestible organic material and larger, rich food items, blurring the macrophage-microphage dichotomy that applies well to larger worms.

Burrowing behavior in mud and sand of morphologically divergent polychaete species (Annelida: Orbiniidae).

Muddy and sandy sediments have different physical properties. Muds are cohesive elastic solids, whereas granular beach sands are non-cohesive porous media. Infaunal organisms such as worms that burrow through sediments therefore face different mechanical challenges that potentially lead to a variety of burrowing strategies and morphologies. In this study we compared three morphologically distinct polychaete species representing different clades in the family Orbiniidae and related differences in their burrowing behaviors and morphologies to their natural environments (mud or sand). Worms burrowed in transparent analogs for muds and sands, and kinematic analysis showed differences both among species and between materials. Leitoscoloplos pugettensis lives in mud and burrows by fracture, using its pointed head to concentrate stress at the tip of the burrow. Naineris dendritica lives in sand and uses its broader head that fluctuates in width over a burrowing cycle to decrease backward slipping in sand, potentially preventing burrow collapse. Orbinia johnsoni lives in sand and uses internal body expansions to pack sand grains, another mechanism to prevent burrow collapse. By combining data from species and materials to obtain a broad range of burrowing velocities, we show that burrowing worms control their velocity by increasing or decreasing their burrowing frequency rather than by altering cycle distance as shown previously for crawling earthworms. This study demonstrates how fairly small evolutionary divergences in morphologies and behaviors facilitate locomotion in environments with different physical constraints.

Relating divergence in polychaete musculature to different burrowing behaviors: a study using opheliidae (Annelida).

Divergent morphologies among related species are often correlated with distinct behaviors and habitat uses. Considerable morphological and behavioral differences are found between two major clades within the polychaete family Opheliidae. For instance, Thoracophelia mucronata burrows by peristalsis, whereas Armandia brevis exhibits undulatory burrowing. We investigate the anatomical differences that allow for these distinct burrowing behaviors, then interpret these differences in an evolutionary context using broader phylogenetic (DNA-based) and morphological analyses of Opheliidae and taxa, such as Scalibregmatidae and Polygordiidae. Histological three-dimensional-reconstruction of A. brevis reveals bilateral longitudinal muscle bands as the prominent musculature of the body. Circular muscles are absent; instead oblique muscles act with unilateral contraction of longitudinal muscles to bend the body during undulation. The angle of helical fibers in the cuticle is consistent with the fibers supporting turgidity of the body rather than resisting radial expansion from longitudinal muscle contraction. Circular muscles are present in the anterior of T. mucronata, and they branch away from the body wall to form oblique muscles. Helical fibers in the cuticle are more axially oriented than those in undulatory burrowers, facilitating radial expansion during peristalsis. A transition in musculature accompanies the change in external morphology from the thorax to the abdomen, which has oblique muscles similar to A. brevis. Muscles in the muscular septum, which extends posteriorly to form the injector organ, act in synchrony with the body wall musculature during peristalsis: they contract to push fluid anteriorly and expand the head region following a direct peristaltic wave of the body wall muscles. The septum of A. brevis is much thinner and is presumably used for eversion of a nonmuscular pharynx. Mapping of morphological characters onto the molecular-based phylogeny shows close links between musculature and behavior, but less correlation with habitat.

Meandering worms: mechanics of undulatory burrowing in muds.

Recent work has shown that muddy sediments are elastic solids through which animals extend burrows by fracture, whereas non-cohesive granular sands fluidize around some burrowers. These different mechanical responses are reflected in the morphologies and behaviours of their respective inhabitants. However, Armandia brevis, a mud-burrowing opheliid polychaete, lacks an expansible anterior consistent with fracturing mud, and instead uses undulatory movements similar to those of sandfish lizards that fluidize desert sands. Here, we show that A. brevis neither fractures nor fluidizes sediments, but instead uses a third mechanism, plastically rearranging sediment grains to create a burrow. The curvature of the undulating body fits meander geometry used to describe rivers, and changes in curvature driven by muscle contraction are similar for swimming and burrowing worms, indicating that the same gait is used in both sediments and water. Large calculated friction forces for undulatory burrowers suggest that sediment mechanics affect undulatory and peristaltic burrowers differently; undulatory burrowing may be more effective for small worms that live in sediments not compacted or cohesive enough to extend burrows by fracture.

Validation of three sympatric Thoracophelia species (Annelida: Opheliidae) from Dillon Beach, California using mitochondrial and nuclear DNA sequence data.

Thoracophelia (Annelida, Opheliidae) are burrowing deposit feeders generally found in the mid- to upper intertidal areas of sandy beaches. Thoracophelia mucronata (Treadwell, 1914) is found along the west coast of North America, including at Dillon Beach, CA. Two additional species, Thoracophelia dillonensis (Hartman, 1938) and T. williamsi (Hartman, 1938) were also described from this beach. These three sympatric species have been primarily distinguished by branchial morphology, and efforts to determine the validity of the species have been based on morphological, reproductive and ecological studies. Here we demonstrate using mitochondrial and nuclear DNA sequence data that these three species are valid. Mitochondrial Cytochrome c subunit 1 (COI) sequences show uncorrected interspecific distances of ~9-13%. We found no inter-specific differences in body color or in hemoglobin concentration, but found that reproductive males were pinkish-red in color and had lower hemoglobin concentrations than purplish-red reproductive females.

Anchor ice and benthic disturbance in shallow Antarctic waters: interspecific variation in initiation and propagation of ice crystals.

Sea ice typically forms at the ocean's surface, but given a source of supercooled water, an unusual form of ice--anchor ice--can grow on objects in the water column or at the seafloor. For several decades, ecologists have considered anchor ice to be an important agent of disturbance in the shallow-water benthic communities of McMurdo Sound, Antarctica, and potentially elsewhere in polar seas. Divers have documented anchor ice in the McMurdo communities, and its presence coincides with reduced abundance of the sponge Homaxinella balfourensis, which provides habitat for a diverse assemblage of benthic organisms. However, the mechanism of this disturbance has not been explored. Here we show interspecific differences in anchor-ice formation and propagation characteristics for Antarctic benthic organisms. The sponges H. balfourensis and Suberites caminatus show increased incidence of formation and accelerated spread of ice crystals compared to urchins and sea stars. Anchor ice also forms readily on sediments, from which it can grow and adhere to organisms. Our results are consistent with, and provide a potential first step toward, an explanation for disturbance patterns observed in shallow polar benthic communities. Interspecific differences in ice formation raise questions about how surface tissue characteristics such as surface area, rugosity, and mucus coating affect ice formation on invertebrates.

Energetics of burrowing by the cirratulid polychaete Cirriformia moorei.

Burrowing through marine sediments has been considered to be much more energetically expensive than other forms of locomotion, but previous studies were based solely on external work calculations and lacked an understanding of the mechanical responses of sediments to forces applied by burrowers. Muddy sediments are elastic solids through which worms extend crack-shaped burrows by fracture. Here we present data on energetics of burrowing by Cirriformia moorei. We calculated the external energy per distance traveled from the sum of the work to extend the burrow by fracture and the elastic work done to displace sediment as a worm moves into the newly formed burrow to be 9.7 J kg(-1) m(-1) in gelatin and 64 J kg(-1) m(-1) in sediment, much higher than for running or walking. However, because burrowing worms travel at slow speeds, the increase in metabolic rate due to burrowing is predicted to be small. We tested this prediction by measuring aerobic metabolism (oxygen consumption rates) and anaerobic metabolism (concentrations of the anaerobic metabolite tauropine and the energy-storage molecule phosphocreatine) of C. moorei. None of these components was significantly different between burrowing and resting worms, and the low increases in oxygen consumption rates or tauropine concentrations predicted from external work calculations were within the variability observed across individuals. This result suggests that the energy to burrow, which could come from aerobic or anaerobic sources, is not a substantial component of the total metabolic energy of a worm. Burrowing incurs a low cost per unit of time.

Burrow extension with a proboscis: mechanics of burrowing by the glycerid Hemipodus simplex.

Burrowing marine infauna are morphologically diverse and ecologically important as ecosystem engineers. The polychaetes Nereis virens and Cirriformia moorei extend their burrows by crack propagation. Nereis virens does so by everting its pharynx and C. moorei, lacking an eversible pharynx or proboscis, uses its hydrostatic skeleton to expand its anterior. Both behaviors apply stress to the burrow wall that is amplified at the tip of the crack, which extends by fracture. That two species with such distinct morphologies and life histories both burrow by fracturing sediment suggests that this mechanism may be widespread among burrowers. We tested this hypothesis with the glycerid polychaete Hemipodus simplex, which has an eversible proboscis that is much longer and everts more rapidly than the pharynx of N. virens. When the proboscis is fully everted, the tip flares out wider than the rest of the proboscis, creating a shape and applying a stress distribution similar to that of N. virens and resulting in relatively large forces near the tip of the crack. These forces are larger than necessary to extend the crack by fracture and are surprisingly uncorrelated with the resulting stress amplification at the crack tip, which is also larger than necessary to extend the burrow by fracture. These large forces may plastically deform the mud, allowing the worm to build a semi-permanent burrow. Our results illustrate that similar mechanisms of burrowing are used by morphologically different burrowers.

Mechanics and kinematics of backward burrowing by the polychaete Cirriformia moorei.

The polychaete Cirriformia moorei burrows in muddy sediments by fracture, using its hydrostatic skeleton to expand its anterior region and exert force against its burrow wall to extend a crack. Burrowing occurs in four phases: stretching forward into the burrow, extending the crack anteriorly, thickening the burrowing end to amplify stress at the tip of the crack, and bringing the rest of the body forward as a peristaltic wave travels posteriorly. Here, we show that C. moorei is also able to burrow with its posterior end using a similar mechanism of crack propagation and exhibiting the same four phases of burrowing. Worms burrowed backwards with similar speeds and stress intensity factors as forward burrowing, but were thinner and less blunt and did not slip as far away from the crack tip between cycles of burrowing. The anterior end is more muscular and rigid, and differences in body shapes are consistent with having reduce musculature to dilate the posterior segments while burrowing. Backward burrowing provides a unique opportunity to study the effects of morphology on burrowing mechanics within the same species under identical conditions.