Eurydice Pulchra Research Articles

The circadian clock gene bmal1 is necessary for co-ordinated circatidal rhythms in the marine isopod Eurydice pulchra (Leach).

Circadian clocks in terrestrial animals are encoded by molecular feedback loops involving the negative regulators PERIOD, TIMELESS or CRYPTOCHROME2 and positive transcription factors CLOCK and BMAL1/CYCLE. The molecular basis of circatidal (~12.4 hour) or other lunar-mediated cycles (~15 day, ~29 day), widely expressed in coastal organisms, is unknown. Disrupting circadian clockworks does not appear to affect lunar-based rhythms in several organisms that inhabit the shoreline suggesting a molecular independence of the two cycles. Nevertheless, pharmacological inhibition of casein kinase 1 (CK1) that targets PERIOD stability in mammals and flies, affects both circadian and circatidal phenotypes in Eurydice pulchra (Ep), the speckled sea-louse. Here we show that these drug inhibitors of CK1 also affect the phosphorylation of EpCLK and EpBMAL1 and disrupt EpCLK-BMAL1-mediated transcription in Drosophila S2 cells, revealing a potential link between these two positive circadian regulators and circatidal behaviour. We therefore performed dsRNAi knockdown of Epbmal1 as well as the major negative regulator in Eurydice, Epcry2 in animals taken from the wild. Epcry2 and Epbmal1 knockdown disrupted Eurydice's circadian phenotypes of chromatophore dispersion, tim mRNA cycling and the circadian modulation of circatidal swimming, as expected. However, circatidal behaviour was particularly sensitive to Epbmal1 knockdown with consistent effects on the power, amplitude and rhythmicity of the circatidal swimming cycle. Thus, three Eurydice negative circadian regulators, EpCRY2, in addition to EpPER and EpTIM (from a previous study), do not appear to be required for the expression of robust circatidal behaviour, in contrast to the positive regulator EpBMAL1. We suggest a neurogenetic model whereby the positive circadian regulators EpBMAL1-CLK are shared between circadian and circatidal mechanisms in Eurydice but circatidal rhythms require a novel, as yet unknown negative regulator.

Methods for Delivery of dsRNAi Against Canonical Clock Genes and Immunocytodetection of Clock Proteins in Crustacea.

The use of nonclassical model organisms for biological rhythm research has become popular in the last two decades. Here we describe techniques for delivery of dsRNAi molecules to knock down clock gene transcripts in a small intertidal crustacean, Eurydice pulchra, as well as our method for immunodetection of clock proteins in the brain. These methods can be generalized for gene knockdown in any small crustacean or arthropod in which mutagenesis by other methods is neither practical nor possible.

Soft-bottom crustacean fauna from the Turkish coast of the Black and Marmara seas with new records

AbstractIt is becoming increasingly important to monitor zoobenthic biodiversity in seas that are under industrial and anthropogenic pollution pressure, such as the Black Sea and the Marmara Sea. This study covers crustacean species in the Turkish waters of the Black Sea and the Marmara Sea, both of which are closed systems. Sampling was carried out in July–August 2019 and yielded 32 crustacean species from the Black Sea and 77 species from the Marmara Sea. In the Marmara Sea, two species [Kupellonura mediterraneaandLeucon(Macrauloleucon)siphonatus] represent new records for the Turkish waters, and 12 species [Cirolana cranchii,Cumella(Cumella)pygmaea, Cyathura carinata,Cymodoce truncata,Eurydice pulchra,Gammaropsis sophiae,Harpinia truncata,Iphinoe serrata,Iphinoe trispinosa,Liocarcinus pusillus,Nebalia strausiandSynchelidium maculatum] are new to the Marmara Sea. The following species:Gammaropsis palmata,Pontocrates arenarius, andSynchelidium haplochelesare new records for the Black Sea. The order Amphipoda dominated in both seas in terms of the number of species and the number of individuals.

Basal position of two new complete mitochondrial genomes of parasitic Cymothoida (Crustacea: Isopoda) challenges the monophyly of the suborder and phylogeny of the entire order

BackgroundIsopoda is a highly diverse order of crustaceans with more than 10,300 species, many of which are parasitic. Taxonomy and phylogeny within the order, especially those of the suborder Cymothoida Wägele, 1989, are still debated. Mitochondrial (mt) genomes are a useful tool for phylogenetic studies, but their availability for isopods is very limited. To explore these phylogenetic controversies on the mt genomic level and study the mt genome evolution in Isopoda, we sequenced mt genomes of two parasitic isopods, Tachaea chinensis Thielemann, 1910 and Ichthyoxenos japonensis Richardson, 1913, belonging to the suborder Cymothoida, and conducted comparative and phylogenetic mt genomic analyses across Isopoda.ResultsThe complete mt genomes of T. chinensis and I. japonensis were 14,616 bp and 15,440 bp in size, respectively, with the A+T content higher than in other isopods (72.7 and 72.8%, respectively). Both genomes code for 13 protein-coding genes, 21 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and possess a control region (CR). Both are missing a gene from the complete tRNA set: T. chinensis lacks trnS1 and I. japonensis lacks trnI. Both possess unique gene orders among isopods. Within the CR of I. japonensis (284 bp), we identified a repetitive region with four tandem repeats. Phylogenetic analysis based on concatenated nucleotide sequences of 13 protein-coding genes showed that the two parasitic cymothoids clustered together and formed a basal clade within Isopoda. However, another parasitic cymothoid, Gyge ovalis Shiino, 1939, formed a sister group with the suborder Limnoriidea Brandt & Poore in Poore, 2002, whereas two free-living cymothoid species were located in the derived part of the phylogram: Bathynomus sp. formed a sister group with the suborder Sphaeromatidea Wägele, 1989, and Eurydice pulchra Leach, 1815 with a clade including Bathynomus sp., Sphaeromatidea and Valvifera G. O. Sars, 1883.ConclusionsOur results did not recover the suborders Cymothoida and Oniscidea Latreille, 1802 as monophyletic, with parasitic and free-living cymothoidans forming separate clades. Furthermore, two parasitic cymothoidans formed the sister-clade to all other isopods, separated from Epicaridea Latreille, 1825, which challenges currently prevalent isopod phylogeny. Additional mt genomes of parasitic and free-living isopods might confer a sufficient phylogenetic resolution to enable us to resolve their relationships, and ultimately allow us to better understand the evolutionary history of the entire isopod order.

The first complete mitochondrial genome of a parasitic isopod supports Epicaridea Latreille, 1825 as a suborder and reveals the less conservative genome of isopods.

The complete mitochondrial genome sequence of the holoparasitic isopod Gyge ovalis (Shiino, 1939) has been determined. The mitogenome is 14,268 bp in length and contains 34 genes: 13 protein-coding genes, two ribosomal RNA, 19 tRNA and a control region. Three tRNA genes (trnE, trnI and trnS1) are missing. Most of the tRNA genes show secondary structures which derive from the usual cloverleaf pattern except for trnC which is characterised by the loss of the DHU-arm. Compared to the isopod ground pattern and Eurydice pulchra Leach, 1815 (suborder Cymothoida Wägele, 1989), the genome of G. ovalis shows few differences, with changes only around the control region. However, the genome of G. ovalis is very different from that of non-cymothoidan isopods and reveals that the gene order evolution in isopods is less conservative compared to other crustaceans. Phylogenic trees were constructed using maxiumum likelihood and Bayesian inference analyses based on 13 protein-coding genes. The results do not support the placement of G. ovalis with E. pulchra and Bathynomus sp. in the same suborder; rather, G. ovalis appears to have a closer relationship to Ligia oceanica (Linnaeus, 1767), but this result suggests a need for more data and further analysis. Nevertheless, these results cast doubt that Epicaridea Latreille, 1825 can be placed as an infraorder within the suborder Cymothoida, and Epicaridea appears to also deserve subordinal rank. Further development of robust phylogenetic relationships across Isopoda Latreille, 1817 will require more genetic data from a greater diversity of taxa belonging to all isopod suborders.

Characterization, localization and temporal expression of crustacean hyperglycemic hormone (CHH) in the behaviorally rhythmic peracarid crustaceans, Eurydice pulchra (Leach) and Talitrus saltator (Montagu)

Crustacean hyperglycemic hormone (CHH) has been extensively studied in decapod crustaceans where it is known to exert pleiotropic effects, including regulation of blood glucose levels. Hyperglycemia in decapods seems to be temporally gated to coincide with periods of activity, under circadian clock control. Here, we used gene cloning, in situ hybridization and immunohistochemistry to describe the characterization and localization of CHH in two peracarid crustaceans, Eurydice pulchra and Talitrus saltator. We also exploited the robust behavioral rhythmicity of these species to test the hypothesis that CHH mRNA expression would resonate with their circatidal (12.4h) and circadian (24h) behavioral phenotypes. We show that both species express a single CHH transcript in the cerebral ganglia, encoding peptides featuring all expected, conserved characteristics of other CHHs. E. pulchra preproCHH is an amidated 73 amino acid peptide N-terminally flanked by a short, 18 amino acid precursor related peptide (CPRP) whilst the T. saltator prohormone is also amidated but 72 amino acids in length and has a 56 residue CPRP. The localization of both was mapped by immunohistochemistry to the protocerebrum with axon tracts leading to the sinus gland and into the tritocerebrum, with striking similarities to terrestrial isopod species. We substantiated the cellular position of CHH immunoreactive cells by in situ hybridization. Although both species showed robust activity rhythms, neither exhibited rhythmic transcriptional activity indicating that CHH transcription is not likely to be under clock control. These data make a contribution to the inventory of CHHs that is currently lacking for non-decapod species.

Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk.

The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12–12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster.

Dissociation of Circadian and Circatidal Timekeeping in the Marine Crustacean Eurydice pulchra

SummaryBackgroundTidal (12.4 hr) cycles of behavior and physiology adapt intertidal organisms to temporally complex coastal environments, yet their underlying mechanism is unknown. However, the very existence of an independent “circatidal” clock has been disputed, and it has been argued that tidal rhythms arise as a submultiple of a circadian clock, operating in dual oscillators whose outputs are held in antiphase i.e., ∼12.4 hr apart.ResultsWe demonstrate that the intertidal crustacean Eurydice pulchra (Leach) exhibits robust tidal cycles of swimming in parallel to circadian (24 hr) rhythms in behavioral, physiological and molecular phenotypes. Importantly, ∼12.4 hr cycles of swimming are sustained in constant conditions, they can be entrained by suitable stimuli, and they are temperature compensated, thereby meeting the three criteria that define a biological clock. Unexpectedly, tidal rhythms (like circadian rhythms) are sensitive to pharmacological inhibition of Casein kinase 1, suggesting the possibility of shared clock substrates. However, cloning the canonical circadian genes of E. pulchra to provide molecular markers of circadian timing and also reagents to disrupt it by RNAi revealed that environmental and molecular manipulations that confound circadian timing do not affect tidal timing. Thus, competent circadian timing is neither an inevitable nor necessary element of tidal timekeeping.ConclusionsWe demonstrate that tidal rhythms are driven by a dedicated circatidal pacemaker that is distinct from the circadian system of E. pulchra, thereby resolving a long-standing debate regarding the nature of the circatidal mechanism.

Macrofaunal sediment selectivity considerations for beach nourishment programmes

Nowadays, beach nourishment is widely considered as a better alternative compared to the construction of hard structures to protect a sandy coast against detrimental erosive effects, both from an ecological and an engineering perspective. The rare studies conducted on the ecological impact of beach nourishment are short-term, post hoc monitoring investigations of the benthic macrofauna. Little is known of the biological processes during and after nourishment. To allow swift recolonization after nourishment, the characteristics of the nourished beach have to match the habitat demands of the benthic macrofauna. The sediment preference of the key intertidal species Scolelepis squamata, Eurydice pulchra, Bathyporeia pilosa and Bathyporeia sarsi, which dominate many West European sandy beaches, was investigated through laboratory experiments, both in single-species as well as combined-species treatments. While the former aimed at developing guidelines for impact mitigation of beach nourishment, the latter aimed at elucidating the role of biotic interactions in sediment preference. Results of the experiments indicated that B. pilosa and E. pulchra prefer the finest sediment, while B. sarsi had a broader preference and also occurred in medium-coarse sediments. However, the sediment preference of E. pulchra for fine sediments was not confirmed by other field and experimental studies. The polychaete S. squamata had the broadest preference and even showed a high occurrence in coarse sediments that are not naturally occurring on the sandy beaches where the animals were caught for this experiment. However, this polychaete is a cosmopolitan species, not only occurring on fine-grained beaches, but also on coarse-grained beaches worldwide. The preferences imply that beach nourishment with coarse sediment will have a major effect on B. pilosa while effects of coarse sediments on S. squamata will be minor. Finally, interspecific competition with the sympatrically occurring amphipod B. sarsi was found to change the sediment selection of the amphipod B. pilosa towards the coarser sediments where B. sarsi occurred in lower frequencies.

Does beach nourishment have long-term effects on intertidal macroinvertebrate species abundance?

Coastal squeeze is the largest threat for sandy coastal areas. To mitigate seaward threats, erosion and sea level rise, sand nourishment is commonly applied. However, its long-term consequences for macroinvertebrate fauna, critical to most ecosystem services of sandy coasts, are still unknown. Seventeen sandy beaches – nourished and controls – were sampled along a chronosequence to investigate the abundance of four dominant macrofauna species and their relations with nourishment year and relevant coastal environmental variables. Dean's parameter and latitude significantly explained the abundance of the spionid polychaete Scolelepis squamata, Beach Index (BI), sand skewness, beach slope and latitude explained the abundance of the amphipod Haustorius arenarius and Relative Tide Range (RTR), recreation and sand sorting explained the abundance of Bathyporeia sarsi. For Eurydice pulchra, no environmental variable explained its abundance. For H. arenarius, E. pulchra and B. sarsi, there was no relation with nourishment year, indicating that recovery took place within a year after nourishment. Scolelepis squamata initially profited from the nourishment with “over-recolonisation”. This confirms its role as an opportunistic species, thereby altering the initial community structure on a beach after nourishment. We conclude that the responses of the four dominant invertebrates studied in the years following beach nourishment are species specific. This shows the importance of knowing the autecology of the sandy beach macroinvertebrate fauna in order to be able to mitigate the effects of beach nourishment and other environmental impacts.

A novel form of pigment‐dispersing hormone in the central nervous system of the intertidal marine isopod, Eurydice pulchra (leach)

Pigment-dispersing factor (PDF) is well known as a circadian clock output factor, which drives daily activity rhythms in many insects. The role of its homologue, pigment-dispersing hormone (PDH), in the regulation of circadian and/or circatidal rhythmicity in crustaceans is, however, poorly understood. The intertidal isopod crustacean, Eurydice pulchra has well-defined circatidal (12.4-hour) activity rhythms. In this study we show that this runs parallel to a circadian (24-hour) cycle of chromatophore dispersion. As a first step in determining the potential role of PDH in these rhythms, we have identified a novel form of PDH expressed in this species. Because conventional homology cloning was unsuccessful, we employed immuno-identification and Edman microsequencing to determine the primary structure of this peptide. From this, cDNA cloning identified the nucleotide encoding sequence and thus facilitated description of PDH neurons by in situ hybridization and immunohistochemistry. We show them to be morphologically similar to those that co-ordinate circadian activity rhythms in insects. In animals expressing both tidal (activity) and circadian (chromatophore) rhythms, however, there was no evidence for a corresponding periodicity in the expression of pdh transcript, as determined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in Eurydice heads. It is therefore suggested that any role for PDH in daily/tidal timing in Eurydice is not mediated at the transcriptional level, rather rhythms in neurohemal release may be important in such co-ordination.

The Belgian sandy beach ecosystem: a review

AbstractThis paper reviews the available knowledge on sedimentology, hydrodynamics and five major ecosystem components (microphytobenthos, vascular plants, terrestrial arthropods, zoobenthos, and avifauna) of Belgian sandy beaches. It covers the area from the foredunes to the lower foreshore, takes an ecosystem approach to beaches of this specific geographic area. Morphodynamically, Belgian beaches are (ultra‐)dissipative, macrotidal, and wide. Characteristic grain sizes are 160–380 μm. The sand becomes coarser, beach slopes steeper and tidal range smaller towards the south‐west, where beaches have also been frequently reshaped by human interference such as nourishment. Beach organisms are highly adapted to this unique environment and can reach high numbers. We show that even on a heavily populated coastline subjected to intense recreational and development pressure, beaches provide critical, yet threatened, habitats. Vascular plants growing near the drift line, on the dry beach and in the embryonic dunes are mostly short‐lived and thalassochorous; the most common species include sea rocket (Cakile maritima), prickly saltwort (Salsola kali subsp. kali), and sea sandwort (Honckenya peploides). These zones are habitat to a highly diverse suite of terrestrial arthropods of halobiontic, halophilous and haloxene species; prominent members are sandhoppers (Talitrus saltator) and dipterans (flies). Microphytobenthos, mainly diatoms, is an important primary producer on Belgian beaches but is not well known. The meio‐ and macrobenthos of Belgian beaches contains specific assemblages such as the Scolelepis squamata–Eurydice pulchra community of the upper intertidal zone. Birds no longer nest on the beaches itself, but Belgian sandy shores continue to function as important resting and foraging areas for birds such as the sanderling Calidris alba. We identify several human pressures on the beach ecosystems arising from recreation, beach management and fisheries.

Experimental biological effects assessment associated with on-shore brine discharge from the creation of gas storage caverns

Most of the studies on biological and ecological effects associated with brine discharge into the marine environment are related to the operation of desalination plants, for the production of freshwater. In this study we analysed the biological effects of a brine effluent from a completely different source, produced from the lixiviation of rock salt caves, for the creation of natural gas storage caverns. Lethal and sub-lethal endpoints following exposure to the brine were studied in a range of macrofauna species characteristic of the soft and hard bottom habitats in the vicinity of the discharge area, namely the isopod Eurydice pulchra, the annelids Sabellaria alveolata and Ophelia radiata, the sea-urchin Paracentrotus lividus and the bivalve Mytilus galloprovincialis. In a first series of experiments, brine, with salinity above 300, was diluted in distilled water to a salinity value close to that of the seawater in the discharge area (salinity 36) and, surprisingly, none of the exposed species was able to survive or develop into viable larvae. A second series of experiments exposed the species to brine diluted with seawater, simulating more realistic discharge circumstances. All the tested species at all the measured endpoints (adult survival, larval abnormal development, sperm fertilization success) showed negative biological effects in brine solutes always at a lower salinity than that of a salinity control obtained with concentrated seawater. The sub-lethal experiments with larval development of P. lividus, S. alveolata and M. galloprovincialis, and the fertilization success of P. lividus gave EC 50 values for the brine solute with salinity in the range of 40.9–43.5, whereas the EC 50 values for the concentrated seawater were in the range of salinity 44.2–49.0. It is hypothesised that differences in the ionic composition of the brine cause the inability of the species to tolerate the exposure to brine.

Identification of putative crustacean neuropeptides using in silico analyses of publicly accessible expressed sequence tags

The development of expressed sequence tags (ESTs) for crustacean cDNA libraries and their deposition in publicly accessible databases has generated a rich resource for peptide discovery in this commercially and ecologically important arthropod subphylum. Here, we have conducted in silico searches of these databases for unannotated ESTs encoding putative neuropeptide precursors using the BLAST program tblastn, and have predicted the mature forms of the peptides encoded by them. The primary strategy used was to query the database with known decapod prepro-hormone sequences or, in some instances, insect precursor protein sequences. For neuropeptides for which no prepro-hormones are known, the peptides themselves were used as queries. For those peptides expected to originate from a common precursor, the individual sequences were combined, with each peptide flanked by a dibasic processing site and, if amidated, a glycine residue. Using these approaches, 13 unannotated ESTs encoding putative neuropeptide precursors were found. For example, using the first strategy, putative Marsupenaeus japonicus prepro-hormones encoding B-type allatostatins, neuropeptide F (NPF), and orcokinins were identified. Similarly, several Homarus americanus ESTs encoding putative orcokinin precursors were found. In addition to the decapod prepro-hormones, ESTs putatively encoding a NPF isoform and a red pigment concentrating hormone-like peptide were identified from the cladoceran Daphnia magna, as was one EST putatively encoding multiple tachykinin-related peptides from the isopod Eurydice pulchra. Using the second strategy, we identified a Carcinus maenas EST encoding HIGSLYRamide, a peptide recently discovered via mass spectrometry from Cancer productus. Using mass spectral methods we confirmed that this peptide is also present in Carcinus maenas. Collectively over 50 novel crustacean peptides were predicted from the identified ESTs, providing a strong foundation for future investigations of the evolution, regulation and function of these and related molecules in this arthropod taxon.

Macrobenthic community structure of soft-bottom sediments at the Belgian Continental Shelf

Within the frame of different research projects, a large number of sites at the Belgian Continental Shelf (BCS) have been sampled for the macrobenthos between 1994 and 2000. These samples cover a diverse range of habitats: from the sandy beaches to the open sea, from the gullies between the sandbanks to the tops of the sandbanks, and from clay to coarse sandy sediments. To investigate the large-scale spatial distribution of the macrobenthos of the Belgian Continental Shelf, the data of all these research projects—728 samples—were combined and analysed. By means of several multivariate techniques, 10 sample groups with similar macrobenthic assemblage structure were distinguished. Each sample group is found in a particular physico-chemical environment and has a specific species composition. Four sample groups differ drastically, both in habitat and species composition, and are considered to represent four macrobenthic communities: (1) the muddy fine sand Abra alba– Mysella bidentata community is characterized by high densities and diversity; (2) the Nephtys cirrosa community occurs in well-sorted sandy sediments and is characterized by low densities and diversity; (3) very low densities and diversity typify the Ophelia limacina– Glycera lapidum community, which is found in coarse sandy sediments and (4) the Eurydice pulchra– Scolelepis squamata community is typical for the upper intertidal zone of sandy beaches. These macrobenthic communities are not isolated from each other, but are linked through six transitional species assemblages. The transition between the A. alba– M. bidentata community and the N. cirrosa community is characterized by a reduction in the mud content and is dominated by Magelona johnstoni. The transition between the N. cirrosa and the O. limacina– G. lapidum community is distinctive by decreasing densities and coincides with a gradual transition between medium and coarse sandy sediments. From the N. cirrosa to the E. pulchra– S. squamata community, transitional species assemblages related to the transition from the subtidal to the intertidal environment were found. Each community or transitional species assemblages was found over a specific range along the onshore–offshore gradient, four types can be discerned: (1) almost restricted to the near-shore area, but possible wider distribution; (2) distributed over the full onshore–offshore gradient; (3) restricted to the near-shore area and (4) restricted to the sandy beach environment. The diversity pattern on the BCS follows this division, with species rich and poor assemblages in the near-shore area to only species poor assemblages more offshore. The distribution and diversity patterns are linked to the habitat type, distinguished by median grain size and mud content.

Ontogeny of behavioural adaptations in beach crustaceans: some temporal considerations for integrated coastal zone management and conservation

So-called “typical” behavioural responses of coastal animals to particular stimuli have previously been shown often to vary cyclically in phase with diel or tidal cycles in the environment. Less well-studied are differences in the behaviour of adults and juveniles of the same species at the same time of day or tidal state, or in response to the same stimulus. Experimental studies of such differences in behaviour are reviewed and compared for three species of beach crustaceans, namely, the crab Carcinus maenas, the isopod Eurydice pulchra and the amphipod Orchestoidea tuberculata. Juvenile, but not adult, Carcinus will entrain circatidal rhythmicity after exposure to artificial tidal cycles of immersion/emersion; juvenile, but not adult, Eurydice express pronounced free-running circatidal swimming rhythms at neap tides as well as at springs; and, in Orchestoidea, juveniles and adults express patterns of daily locomotor activity that are complementary, both on the shore and in the laboratory. These ontogenetic differences are discussed in relation to distributional and behavioural differences between adults and juveniles in each species, drawing attention to their adaptive significance and wider implications for coastal management and conservation.

Macrobenthic zonation patterns along a morphodynamical continuum of macrotidal, low tide bar/rip and ultra-dissipative sandy beaches

The species composition, densities, biomass and zonation patterns of the macrobenthos of sandy beaches are greatly influenced by the morphodynamics and morphology of the beaches. Macrobenthic zonation patterns along a small-scale morphodynamic gradient, comprising eight Belgian beach sites, were investigated. By taking into account the dimensionless fall velocity ( Ω) and the relative tidal range, the beach sites were ordered along the gradient from the ultra-dissipative beach type (UD) to the low tide bar/rip beach type (LTBR). The resulting beach state index varied between 1.8 and 4.2 and the beach profiles were related with the beaches' morphodynamic state. In total 35 macrobenthic species, mainly polychaetes and crustaceans, were encountered, varying between 19 and 23 species per beach site. The species composition was quite similar among beach sites, with Scolelepis squamata being abundant at all eight sites. Furthermore, the macrobenthic distribution patterns were mainly related to elevation at all beach sites. Some remarkable difference in metrics, largely related to the beach morphodynamics and the consequent hydrodynamics, were found. At the hydrodynamically benign and consequently macrobenthos-rich UD beaches, the highest macrobenthic densities and biomass occurred on the upper beach, while at the hydrodynamically harsh and thus macrobenthos-poor LTBR beaches, the maximum densities and biomass occurred lower on the beach. Species, typically occurring on the upper UD beaches, such as Eurydice pulchra, S. squamata, and Bathyporeia sarsi, were restricted to the sub-optimal middle and lower beach zone at LTBR beaches. Only Bathyporeia pilosa was found on the upper beach of both UD and LTBR beaches. The more robust polychaete Ophelia rathkei and the interstitial polychaete Hesionides arenaria were exclusively found in the hydrodynamically harsh conditions of the middle LTBR beach zone.

Circatidal variability in the behavioural responses of a sandbeach isopod Eurydice pulchra (Leach) to orientational cues

Spontaneous, phasic changes in the behavioural responses of freshly collected Eurydice pulchra (Leach) (Isopoda;Crustacea) to light and water flow were studied in the laboratory and related to what is known of the behaviour of this isopod when free swimming. During the expected flood tide period, isopods in laboratory experiments generally expressed positive phototaxis, high photokinesis and negative rheotaxis. On the shore at that time they usually move upshore in the surface waters of the surf zone. In the laboratory, a spontaneous change in rheotaxis behaviour was recorded at the time of the expected high tide and the isopods tended to stem water currents, presumably associated with the avoidance of stranding on the shore. During the expected ebb tide period, isopods studied in the laboratory showed negative phototaxis, low photokinesis and a resumption of negative rheotaxis, when on the shore they migrate down in the water column and so downshore in the undertow. When held with sand in a test tank, E. pulchra burrowed before expected low tide, away from a directional light source. If, however, the isopods were not able to reburrow due to the absence of sand in laboratory experiments, a return to photopositive swimming was recorded during the expected ebb, behaviour which on the shore would return the individuals to the water's edge. Rhythmic responsiveness of E. pulchra to directional cues appears to fine-tune the tidal swimming pattern of this isopod, inducing temporal changes in behaviour which control both vertical and horizontal distribution on the shore.

The diurnal modulation of the circatidal activity rhythm by feeding in the isopodeurydice pulchra

The diurnal modulation of circatidal activity was studied in freshly caught Eurydice pulchra of different feeding states. Apparently fully fed specimens displayed definite circadian modulation, with greatest tidal activity at the time of expected night‐time high tides, and apparently starved animals showed no circadian variation. Circatidal activity was also entrained in the laboratory using cycles of artificial agitation and the subsequent pattern of free‐running rhythmicity was studied. Animals fed prior to entrainment displayed definite diurnal modulation of the pattern of ciractidal swimming activity. Those fed after the entrainment regime, or not at all, displayed no apparent diurnal inequality of tidal activity peaks. No relationship between diurnal inequality and phototactic behaviour of freshly collected specimens was detected.

An entrainment model for semilunar rhythmic swimming behaviour in the marine isopod Eurydice pulchra Leach

Entrainment experiments have been carried out with geographically widely separated populations of the sand beach isopod Eurydice pulchra Leach subjected to periods of simulated tidal agitation imposed concurrently with a 24-h light: dark (L: D) cycle. Circatidal swimming rhythms of greatest amplitude were induced when agitation was applied with the subjective timing, within the L: D cycle, of local spring high tides. This occurred in a normal L: D regime and also when the L: D regime was phase shifted through 90°. Animals previously maintained in constant darkness (D: D) and subsequently exposed to simulated tidal disturbance at various times in constant darkness were unable to modulate the amplitude of circatidal swimming activity. Isopods previously maintained in a normal L: D cycle and subsequently subjected to artificial tidal agitation in constant darkness were, however, able to modulate circatidal activity. This indicates that E. pulchra is capable of detecting tidal agitation and daily light cues and using them in conjunction with its circadian “clock” to modulate its endogenous circatidal rhythmicity. The free-running semilunar rhythm of swimming activity entrained only when the timing of agitation within the day/night cycle mimicked the pattern of local spring high tides. Agitation with the timing of neap high tides entrained no free-running circa-semilunar activity pattern.