Barnacle Shells Research Articles

Heterogeneous shell growth of the neustonic goose barnacle Lepas anserifera

Floating materials of both natural and anthropogenic origin affect marine ecosystems and human economic activities. Although the tracking of floating materials is important to manage the economic risks, it is difficult to trace them back to the events of origin, such as tsunamis and underwater volcanic eruptions. The gooseneck barnacle Lepas anserifera, a rapid colonizer in pelagic environments, is a potential “natural logger” of floating materials. In this study, we performed temperature-controlled culture experiments and growth line staining in the laboratory to quantify the growth increments of shells (scutum and tergum) consisting the capitulum of L. anserifera separately, and to examine the effects of the temperature on their growth. Following calcein staining, the growth lines of L. anserifera were visualized under a fluorescent microscope, and gross (capitular length and width) and individual (scutum and tergum) shell growth were compared. Shells grew in twice as much in the capitular length direction than in the capitular width direction owing to the larger growth increases in the scutum than in the tergum. Growth increments were unaffected by temperatures in the range from 20°C to 30°C, although the growth appeared to slow down in September and October compared with August. The stable oxygen isotope composition (δ18O) of the shells represented the water temperature as previously known, and the present results showed that 18O enriched in scutum than tergum in most cases. Further understanding for the biomineralization process of barnacles is required for the precise application of environmental proxies in barnacle shells.

Cleaning Equipment Surface with Chemical Solution to Dissolve and Remove Barnacle Shells

Cleaning Equipment Surface with Chemical Solution to Dissolve and Remove Barnacle Shells

A synopsis of the Tanaidacea (Crustacea: Peracarida) of Singapore, with a review of tanaidacean diversity in Southeast Asia and the South China Sea

In the first comprehensive taxonomic study of the tanaidacean fauna of Singapore, a total of 23 species belonging to two suborders, three superfamilies, 13 families and 22 genera were identified from approximately 2,400 specimens, including 11 species that are possibly new to science. This material was collected from various localities in Singapore waters, from the intertidal zone to subtidal habitats up to 91 m in depth. Many species exhibited a narrow bathymetric distribution and strong affiliation to certain habitats or microhabitats such as mudflats, coral reefs, and barnacle shells on rocky shores. The Singapore Strait yielded 20 tanaidacean species, almost thrice the number of species (7) present in the Johor Straits. Three species were confined to the brackish waters in the inner parts of the Johor Straits and Sungei Pandan. Furthermore, a total of 153 tanaidacean species has now been recorded from Southeast Asia and the South China Sea. The presence of the family Numbakullidae in this region is also documented for the first time.

New genes helped acorn barnacles adapt to a sessile lifestyle.

Barnacles are the only sessile lineages among crustaceans, and their sessile life begins with the settlement of swimming larvae (cyprids) and the formation of protective shells. These processes are crucial for adaptation to a sessile lifestyle, but the underlying molecular mechanisms remain poorly understood. While investigating these mechanisms in the acorn barnacle, Amphibalanus amphitrite, we discovered a new gene, bcs-6, which is involved in the energy metabolism of cyprid settlement and originated from a transposon by acquiring the promoter and cis-regulatory element. Unlike mollusks, the barnacle shell comprises alternate layers of chitin and calcite and requires another new gene, bsf, which generates silk-like fibers that efficiently bind chitin and aggregate calcite in the aquatic environment. Our findings highlight the importance of exploring new genes in unique adaptative scenarios, and the results will provide important insights into gene origin and material development.

Potential of processed barnacles shell as soil conditioner using mung bean plants (Vigna radiata L.) growth

Barnacles as periphyte animals often found clinging on the ship’s hull, being unutilized waste that presents in such abundance. Calcium carbonate in barnacle shells would increase the chance of degrading microbial community survival in the soil, indirectly increasing soil fertility. The processing of barnacle shells to be used as a calcium carbonate supplement involves harvesting, drying, powdering, sterilizing, and mixing it to the soil. We want to observe comparison of the performance of control group (K), processed barnacle shells (T), industrial 16-16-16 NPK (Nitrogen, Phosphate, Potassium) fertilizer (P), and the combination of processed barnacle shell with 16-16-16 NPK fertilizer (TP) supplementation to the soil as planting media by the daily temperature, pH, and moisture fluctuation, besides the measurement of the growth of mung bean plant, Vigna radiata L. (VIMA 5 variant) regarding their application to the soil. The growth of V. radiata L. is measured by epicotyl length, number of leaves, and total wet mass increase of each plant individual. Vigna radiata L. in the control group shows the best growth measured by parameters listed above. However, ANOVA analysis shows statistical significance on the correlation of types of soil treatment (K, T, P, or TP supplementation) to daily moisture fluctuation.

Dopamine-Based Copolymer Bottlebrushes for Functional Adhesives: Synthesis, Characterization, and Applications in Surface Engineering of Antifouling Polyethylene.

Nonpolar materials like polyolefins are notoriously challenging substrates for surface modification. However, this challenge is not observed in nature. Barnacle shells and mussels, for example, utilize catechol-based chemistry to fasten themselves onto all kinds of materials, such as boat hulls or plastic waste. Here, a design is proposed, synthesized, and demonstrated for a class of catechol-containing copolymers (terpolymers) for surface functionalization of polyolefins. Dopamine methacrylamide (DOMA), a catechol-containing monomer, is incorporated into a polymer chain together with methyl methacrylate (MMA) and 2-(2-bromoisobutyryloxy)ethyl methacrylate (BIEM). DOMA serves as adhesion points, BIEM provides functional sites for subsequent "grafting from" reactions, and MMA provides the possibility for concentration and conformation adjustment. First, the adhesive capabilities of DOMA are demonstrated by varying its content in the copolymer. Then, terpolymers are spin-coated on model Si substrates. Subsequently, the atom transfer initiator (ATRP) initiating group is used to graft a poly(methyl methacrylate) (PMMA) layer from the copolymers, with 40% DOMA content providing a coherent PMMA film. To demonstrate functionalization on a polyolefin substrate, the copolymer is spin-coated on high-density polyethylene (HDPE) substrates. A POEGMA layer is grafted from the ATRP initiator sites on the terpolymer chain on the HDPE films to provide antifouling characteristics. Static contact angle values and Fourier transform infrared (FTIR) spectra confirm the presence of POEGMA on the HDPE substrate. Finally, the anticipated antifouling functionality of grafted POEGMA is demonstrated by observing the inhibition of nonspecific adsorption of the fluorescein-modified bovine serum albumin (BSA) protein. The poly(oligoethylene glycol methacrylate) POEGMA layers grafted on 30% DOMA-containing copolymers on HDPE show optimal antifouling performance exhibiting a 95% reduction of BSA fluorescence compared to nonfunctionalized and surface-fouled polyethylene. These results demonstrate the successful utilization of catechol-based materials for functionalizing polyolefin surfaces.

Convergent evolution of barnacles and molluscs sheds lights in origin and diversification of calcareous shell and sessile lifestyle.

The calcareous shell and sessile lifestyle are the representative phenotypes of many molluscs, which happen to be present in barnacles, a group of unique crustaceans. The origin of these phenotypes is unclear, but it may be embodied in the convergent genetics of such distant groups (interphylum). Herein, we perform comprehensive comparative genomics analysis in barnacles and molluscs, and reveal a genome-wide strong convergent molecular evolution between them, including coexpansion of biomineralization and organic matrix genes for shell formation, and origination of lineage-specific orphan genes for settlement. Notably, the expanded biomineralization gene encoding alkaline phosphatase evolves a novel, highly conserved motif that may trigger the origin of barnacle shell formation. Unlike molluscs, barnacles adopt novel organic matrices and cement proteins for shell formation and settlement, respectively, and their calcareous shells have potentially originated from the cuticle system of crustaceans. Therefore, our study corroborates the idea that selection pressures driving convergent evolution may strongly act in organisms inhabiting similar environments regardless of phylogenetic distance. The convergence signatures shed light on the origin of the shell and sessile lifestyle of barnacles and molluscs. In addition, notable non-convergence signatures are also present and may contribute to morphological and functional specificities.

Occurrence and Habitat Characteristics of Aurelia sp. Polyps in a High-Latitude Fjord

Causes and consequences of jellyfish bloom formation are subject to controversial discussions worldwide. While medusae have been studied to a broader extent, the knowledge on polyp stages of scyphozoans is limited thus hampering reliable prediction of jellyfish bloom formation. This study describes the occurrence, abundance, habitat characteristics and interactions of scyphozoan Aurelia sp. polyp colonies with other fouling organisms in intertidal and subtidal sectors of Trondheimsfjorden (Norway). In total, 982 polyps were found on 70 substrata of varying material types during a field survey in spring-summer 2018 along a longitudinal gradient within and outside Trondheimsfjorden. The polyps were identified as Aurelia sp. based on molecular species identification. Most polyps were found in bays with macroalgae canopy on the down facing side of artificial and natural substrata (rocks, concrete, iron) and inside rock cracks. Polyp microhabitats included the surface of Ascidia mentula (solitary ascidian), increments of Pomatoceros triqueter (polychete) tubes and dead Balanus balanoides (barnacle) shells. Based on the deployment of settling plates, abundance of Aurelia sp. polyps ranged from 1.2 (± 0.7) to 0.12 (± 0.07) polyps cm–2. Settlement occurred either directly on the PVC settling plates or as epibionts on the ascidian Ascidia mentula and on barnacle shells of Balanus balanoides. This study provides insights into the potential of local Aurelia sp. polyps contributing to the seasonal occurrence and abundance of Aurelia sp. in fjord systems, where intensive blooms occur annually.

Marine flatworms (Platyhelminthes: Polycladida) found in empty barnacle shells, including a new species, from southern Mexican Pacific.

The association of some species of polyclads of the suborder Acotylea with barnacles has been recorded worldwide. In contrast, no species of the suborder Cotylea has been recorded present in these crustaceans. The aim of this work is to describe the species of polyclads found in the empty barnacle shells on the coasts of Oaxaca, southern Mexican Pacific. Thirty polyclad specimens in the barnacles' empty shells were collected, corresponding to two species. A new species of acotylean, Stylochus mistus sp. nov., was found in Paraconcavus pacificus; and the cotylean Prosthiostomum cf. parvicelis in Megabalanus peninsularis. Both species of polyclads are considered to use the shells of Megabalanus and Paraconcavus as an occasional shelter site; however, it is not ruled out that these species feed on these barnacles, as polyclads normally prey on the communities they inhabit. This work records for the first time the presence of a cotylean polyclad in barnacles, as well as the first record of the polyclad genera Stylochus and Prosthiostomum from the southern Mexican Pacific.

Polydimethylsiloxane Microdomains Formation at the Polythiourethane/Air Interface and Its Influence on Barnacle Release.

In this study, polydimethylsiloxane (PDMS)/polythiourethane (PTU) composite reinforced with tetrapodal shaped micro-nano ZnO particles (t-ZnO) was successfully produced by a versatile, industrially applicable polymer blending process. On the surface of this composite, PDMS is distributed in the form of microdomains embedded in a PTU matrix. The composite inherited not only good mechanical properties originating from PTU but also promising fouling-release (FR) properties due to the presence of PDMS on the surface. It was shown that the preferential segregation of PDMS domains at the polymer/air interface could be attributed to the difference in the surface free energy of PDMS and PTU. The PDMS microdomains at the PTU/air interface significantly reduced the barnacle adhesion strength on the composite. Both the pseudo- and natural barnacle adhesion strength on the composite was approximately 0.1 MPa, similar to that on pure PDMS. The pseudo-barnacle adhesion on reference surfaces AlMg3 and PTU reached approximately 4 and 6 MPa, respectively. Natural barnacles could not be removed intact from AlMg3 and PTU surfaces without breaking the shell, indicating that the adhesion strength was higher than the mechanical strength of a barnacle shell (approximately 0.4 MPa). The integrity of PDMS microdomains was maintained after 12 months of immersion in seawater and barnacle removal. No surface deteriorations were found. In short, the composite showed excellent potential as a long-term stable FR coating for marine applications.

Role of the Organic Matter in the Structural Organization of Giant Barnacle Austromegabalanus Psittacus Shell from the Micro- to Nanoscale

This work is a detailed study of the giant barnacle shell micro-nano-structure and the relationship between the organic matrix and the mineral. We have studied the composition of the organic matrix...

Ichnodiversity and bathymetric range of microbioerosion traces in polar barnacles of Svalbard

This first comprehensive investigation of microbioerosion traces in polar barnacles addresses two bathymetrical transects from the intertidal down to subtidal water depths in two different carbonate factories in the Svalbard Archipelago: the bay Mosselbukta and the ocean bank Bjornoy-Banken. Scanning electron microscopy of epoxy resin casts of barnacle shells yielded 20 different microendolithic bioerosion traces, probably produced by cyanobacteria (three), chlorophytes (two), rhodophytes (one), sponges (one), foraminifera (three), fungi (nine) and bacteria (one). The lowest ichnodiversity in both locations was observed in the shallow euphotic zone and is likely a result of strong temperature fluctuations, extreme seasonality of light levels and episodic sea-ice cover. At 25–150 m water depth, the ichnodiversity remains relatively constant (9–13 ichnospecies), albeit with differing ichnospecies composition, generally dominated by borings from chlorophytes and fungi. Ichnotaxa at Mosselbukta and Bjornoy-Banken were similar in numbers but differed in abundance and slightly also in ichnospecies composition. Statistical tests indicate that water depth (affecting the availability of light) is the most significant driver for the development of different microbioerosion trace assemblages across the bathymetrical transects. In contrast, no significant differences in ichnodisparity were found, indicating a comparable suite of architectural designs of the micro-borings throughout bathymetry and location. The comparison of our results with literature data confirms a decrease in ichnodiversity from lower to higher latitudes, although targeted bioerosion analyses from other polar environments are needed to gain a more complete picture of the role of bioerosion in polar carbonate factories.

Reproductive Aggregations of Dynoides dentisinus (Crustacea: Peracarida), an Intertidal Isopod with Remarkable Sexual Dimorphism.

AbstractThe intertidal isopod Dynoides dentisinus is a sexually dimorphic species; males are much larger than females and have a large, horn-like pleonal process (hereafter referred to as a "horn") and large, posteriorly extended uropods. Here, we investigated the function of these structures with regard to their mating system. Behavioral interactions were observed between a male occupying a small tube (resident) and a newly introduced individual (a female or male visitor). When the visitor was male, the resident repeatedly struck the visitor with his horn; each swing was accompanied by a short sound produced by stridulation. The resident also used his uropods to strike the male visitor and then rejected the visitor. The resident struck the female visitor in a similar fashion but eventually accepted her into the tube; during this process, the resident frequently emitted stridulatory sounds. Our field survey found that the members shared a single shelter (barnacle shell) containing groups composed of several males and females. This suggests that the mating system of this species is polygynandry. The number of females in a single barnacle shell was positively associated with the basal diameter of the barnacle shell. However, the number of females was not associated with the body size, horn size, or uropod size of the largest male in the barnacle shell. These results suggest that male body size, horns, and uropods might have evolved as weapons through male-male competition for large barnacle habitats and more females, but that they have not evolved as ornaments via female choice.

Unique Post-telemetry Recapture Enables Development of Multi-Element Isoscapes From Barnacle Shell for Retracing Host Movement

Many ecological investigations rely on understanding the movement of animals through marine environments. Most available tracking techniques are invasive (e.g., tissue sampling) and require extensive effort and/or cost (e.g., capture-mark-recapture or satellite telemetry). The isotopic compositions of barnacle shells (delta C-13 and delta O-18) are known to record the ambient water temperature and salinity conditions in which they grew. Thus, isotopic analysis of barnacles on animals or objects has the potential to yield information about their movement between water bodies of varying isotopic properties. We present, for the first time, isotopic data for barnacle shell samples that grew on a satellite-tracked sea turtle host. The satellite telemetry record, together with documented barnacle growth rates, allowed for sequential samples from individual barnacle shells to be assigned a specific time and location for direct comparison of isotope values to environmental conditions. We developed models that allow barnacle shell delta C-13 and delta O-18 to be linked, with a high degree of predictability, to sea surface temperature (SST) and salinity (SSS). Our sea turtle case study demonstrated how these models can be used to create isoscapes, allowing hosts to be tracked in space and time at higher resolution than most attempts to use soft-tissue isotopes for a similar purpose, and at considerably lower cost than satellite telemetry. The conceptual advance presented here could be applied widely to understand the movement of any animal or object that carries hitchhiking barnacles.

Uranium isotope compositions of biogenic carbonates – Implications for U uptake in shells and the application of the paleo-ocean oxygenation proxy

The application of U isotopes in carbonates as a paleo-ocean oxygenation proxy is based on the critical assumption that the calcareous shell-building organisms incorporate U into their shells without fractionation relative to the U isotopic composition of ambient seawater. Recent studies claim a small, but resolvable, isotopic offset during abiotic and biogenic aragonite precipitation, whereas no isotope fractionation has been recorded during calcite precipitation. Although aragonite is meta-stable and not preserved over geological timescales (>1 Myr) and U precipitates during diagenesis, the U isotope composition of biogenic aragonite is important because aragonite precipitation is an important U sink to carbonate sediments. In contrast, low-magnesium calcite (LMC) is preserved over geological timescales and may provide a reliable fingerprint of ancient ocean chemistry. Therefore, a more general study is needed that compares U isotope compositions of primary marine biogenic carbonate precipitates.We report the U isotope compositions of 32 modern samples from geographically distinct localities in the Atlantic Ocean including corals (Scleractinia, Octocorallia), brachiopods (Articulata), molluscs (Tellina Listeri, Codahia Obicularis) and barnacles as well as one fossil mollusc. These samples reflect variable primary minerals, water temperatures, water depths, pH-values of ambient water, and U concentrations. Several seawater samples have also been measured to compare our methods with those of previously published studies.The analyzed modern corals and brachiopods display U isotopic compositions that are indistinguishable from modern seawater. This suggests that these carbonates have the potential to faithfully record the U isotopic composition of the surrounding seawater in which they form. The analyzed brachiopods are of particular interest as they are composed of the calcium carbonate polymorph LMC that is stable over geological timescales. While this study shows for the first time that LMC phases are robust targets in ancient samples, their low U abundance presents analytical challenges for precise U isotope analyses. We also show that two barnacle shells collected with ambient seawater have U isotopic compositions that are both lighter and heavier than the ambient seawater. The mechanism to explain this offset is not determined, but it demonstrates that at least barnacle shells are not representative of the seawater in which they last lived. Two of three partially fossilized mollusc shells also show resolvable offsets from seawater, likely indicating secondary processes that are known to shift or fractionate U isotopes. Collectively, our new data indicate that: 1) aragonite delivers U with a seawater composition to carbonate sediments, and 2) LMC shells of brachiopods that are stable over geological timescales may be more suitable for reconstructing the U isotope composition of ancient oceans.

Heavy Metals in Barnacles Balanus Sp.: From Biomonitoring to Coastal Management

This study determined the levels of Cd, Cu, Fe, Ni, and Zn in the soft tissues and shells of barnacles (Balanus sp.) collected from the Kuala Juru, Sebatu, Sg. Ayam and Kg. Pasir Puteh, Peninsular Malaysia, obtained in 2008. The metal concentrations (μg/g dry weight) in the soft tissues of Balanus sp. received from four sites in Peninsular Malaysia ranged from 2.93-4.17 for Cd, 20.2-92.5 for Cu, 480-1193 for Fe, 6.40-18.0 for Ni, and 224-414 for Zn. Based on the present data, the trace metal bioavailability’s to the Balanus in all the populations, especially in a known polluted site at Kg. Pasir Puteh is low. Overall, this preliminary baseline data can be used for regular ecological monitoring for the effective management of the coastal area in Malaysia.

Global oxygen isoscapes for barnacle shells: Application for tracing movement in oceans

It is helpful to understand the movement of animals and objects to inform species conservation and broader environmental management (e.g. by identifying the origin of marine debris). Tagging techniques are limited to investigations of future movement (e.g. after a tag has been applied), with no ability to understand where an animal or object has come from prior to encounter. However, studies that apply chemical techniques are able to address questions about historical movement prior to encounter, particularly through the analysis of stable isotopes from the tissues of migrating animals, or from barnacle shells that attach to migrating hosts. Barnacle shell isotope analysis is a promising technique that could provide a new understanding of the ecology of migrating marine fauna, or additionally the origin of marine debris. Here we use global datasets to assess the applicability of barnacle shell isotope techniques for identifying the origin and travel pathways of animals and objects that carry hitchhiking barnacles. We present the first global isoscapes for barnacle shell calcite, using these to identify areas that are likely to offer the finest spatial resolution for this application. We further demonstrate how isoscapes can be applied to back-trace animal migrations using real-world migration case studies of sea turtles and whales. We demonstrate that coastal areas and mid-latitude oceanic regions are likely to offer the best spatial resolution, and that migration pathways are able to be identified from successive barnacle shell samples. We expect that this work will allow for more efficient and precise future applications of barnacle shell isotope analyses to trace the movement and origin of barnacle hosts through marine waters.

Strontium Isotope Stratigraphy and the thermophilic fossil fauna from the middle Miocene of the East Pisco Basin (Peru)

New age estimates obtained via Strontium Isotope (87Sr/86Sr) Stratigraphy and new paleoclimatic data are here presented for the Miocene Chilcatay and Pisco formations exposed in the East Pisco Basin, an Andean forearc basin of southern Peru, which is renowned worldwide for its exceptional content of fossil marine vertebrates. Mollusk and barnacle shells, carbonate nodules, and shark teeth were collected along three stratigraphic sections for applying Strontium Isotope Stratigraphy on both carbonates and phosphates. To avoid diagenetic biases, mollusk and barnacle shells were analyzed in detail by means of optical and scanning electron microscopy, cathodoluminescence, and inductively coupled plasma-optical emission spectrometry, whereas only the enameloid from the best-preserved shark teeth was sampled. The obtained 87Sr/86Sr ages confirm a late early Miocene (Burdigalian) age for the Chilcatay strata, and reveal middle Miocene (Langhian to Serravallian) ages for the lower Pisco unit (i.e., the P0 sequence) – a result that matches the relatively archaic aspect of its cetacean fossil assemblage. New and literature data about the fossil assemblage of the lower Pisco beds highlight the presence of several thermophilic invertebrates and vertebrates, thus suggesting a warm-water, tropical paleoenvironment for this middle Miocene sequence. Such a paleoenvironmental scenario recalls the warm conditions associated with the Chilcatay Formation, rather than the cooler setting inferred for the remainder of the Pisco Formation (i.e., the P1 and P2 sequences). This pattern likely reflects the late Miocene trend of global cooling, as well as a middle to early late Miocene strengthening of the Humboldt Current.

Substrate-controlled naticid gastropod drilling predation on recent barnacles from Chandipur, India and its Palaeontological implications

ABSTRACT The study of drilling predation has been largely limited to molluscs. Herein, we report drilling predation on Recent acorn barnacles by gastropods from Chandipur, eastern India. The aspects of predator-prey relation dealt here are the size and site preference of the predator; the interrelationship between barnacle shell morphology (e.g., shape, external ornamentation) and drilling intensity; and influence of substrate type on body size and shape of barnacles, as well as predation pressure on it. The study reveals that, barnacles are drilled by the naticid gastropods, which is otherwise an uncommon interaction. Drilling intensity on the barnacles attached to a semi-infaunal bivalve, Timoclea imbricata, is exceptionally high and is comparable to the drilling intensity on the host bivalve. Otherwise, predation pressure is very low on barnacles, supporting previous literature. The study implies that unique live-live associations can create opportunities such that even an unusual predator (e.g., Naticidae) can successfully feed on a novel prey (e.g., barnacles), suggesting dietary opportunism. Studies dealing with the fossil record of drillholes should consider these different ecological (e.g., prey selectivity by the predator, host selection by the epizoan, role of unusual predator on an uncommon prey) aspects of drilling predation on barnacles through space and time.

Distinguishing between sea turtle foraging areas using stable isotopes from commensal barnacle shells

Understanding the movement behaviour of marine megafauna within and between habitats is valuable for informing conservation management, particularly for threatened species. Stable isotope analyses of soft-tissues have been used to understand these parameters in sea turtles, usually relying on concurrent satellite telemetry at high cost. Barnacles that grow on sea turtles have been shown to offer a source of isotopic history that reflects the temperature and salinity of the water in which the host animal has been. We used a novel method that combines barnacle growth rates and stable isotope analysis of barnacle shells (δ18O and δ13C) as predictors of home area for foraging sea turtles. We showed high success rates in assigning turtles to foraging areas in Queensland, Australia, based on isotope ratios from the shells of the barnacles that were attached to them (86–94% when areas were separated by >400 km). This method could be used to understand foraging distribution, migration distances and the habitat use of nesting turtles throughout the world, benefiting conservation and management of these threatened species and may be applied to other taxa that carry hitchhiking barnacles through oceans or estuaries.