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The evolutionary history of crustacean segmentation: a fossil-based perspective.

The evolution of segmentation in Crustacea, that is, the formation of sclerotized and jointed body somites and arrangement of somites into tagmata, is viewed in light of historical traits and functional constraints. The set of Early to Late Cambrian 'Orsten' arthropods have informed our current views of crustacean evolution considerably. These three-dimensionally preserved fossils document ancient morphologies, as opposed to purely hypothetical models and, because of the unusual preservation of larval stages, provide us with unparalleled insight into the morphogenesis of body somites and their structural equipment. The variety of evolutionary levels represented in the 'Orsten' including lobopodians, tardigrades, and pentastomids also allows phylogenetic interpretations far beyond the Crustacea. The 'Orsten' evidence and data from representatives of the Lower Cambrian Chengjiang biota in southwestern China, including phylogenetically earlier forms, form the major source of our morphology-based review of structural and functional developments that led toward the Crustacea. The principal strategy of arthropods is the simultaneous development of head somites, as expressed in a basal "head larva," and a successive addition of postcephalic somites from a preterminal budding zone with progressive maturation of metameric structures. This can be recognized in the developmental patterns of extant and fossil representatives of several euarthropod taxa, particularly crustaceans, trilobites, and chelicerates (at least basally). The development of these taxa points to an early somite-poor and free-living hatching stage. Embryonic development to a late stage within an egg, as occurring in recent onychophorans and certain in-group euarthropods, is regarded as achieved several times convergently.

Animals↗

Phylogenetic comparison of serotonin-immunoreactive neurons in representatives of the Chilopoda, Diplopoda, and Chelicerata: implications for arthropod relationships.

The phylogenetic relationships within the Arthropoda have been discussed controversially for more than a century. Comparative studies on structure and development of the nervous system have contributed important arguments to this discussion. Arthropods have individually identifiable neurons that can be used as characters in phylogenetic studies. In the present report, the arrangement of serotonin-immunoreactive neurons in the ventral nerve cord was examined in seven representatives of the Chelicerata, Chilopoda, and Diplopoda. The goal of this analysis was to determine whether number, arrangement, and axonal morphology of the serotonergic neurons in these groups are similar to the pattern found in representatives of the Hexapoda and Crustacea, as explored in a previous study. The results indicate that the pattern in the seven species examined here does not correspond to that present in the Hexapoda and Crustacea. In particular, the pattern in Chilopoda and Diplopoda is clearly different from that of the Hexapoda. The hexapodan pattern most closely resembles that of the Crustacea. These findings are discussed with regard to recent reports on the mechanisms of neurogenesis in these taxa. Furthermore, the proposed ground patterns of the various groups are reconstructed and the characters are plotted on two competing hypotheses of arthropod phylogeny, the traditional Tracheata hypothesis and an alternative hypothesis derived from molecular and recent morphological data, the Tetraconata concept. The data discussed in this article moderately support the Tetraconata hypothesis.

Animals↗

Sex steroid receptor evolution and signalling in aquatic invertebrates.

In vertebrate reproductive endocrinology sex steroids play a pivotal role via binding to receptors. However, information on the origin and relevance of sex steroids in invertebrates is limited. This review highlights current literature on steroid receptors in aquatic invertebrates and reports on some new findings. It has been shown that invertebrates of the deuterostome clade, such as Acrania and Echinodermata, respond to estrogens and androgens and, at least in Branchiostoma, an estrogen receptor has been cloned. Within the protostomes, most findings are related to aquatic molluscs. Sex steroid receptor-like proteins are abundant in gastropods, bivalves and cephalopods and also sex hormone signalling shows partial similarity to the deuterostomes. In ecdysozoans, however, the impact of sex steroids is still a matter of debate even though there is evidence on the presence of estrogen receptor-like proteins in Crustacea and on physiological effects of estrogens in both Nematoda and Crustacea. Recent findings suggest the presence of an estrogen receptor alpha-like protein of unclear physiological role in Gammarus fossarum (Crustacea). Binding studies revealed the crustacean Hyalella azteca to possess specific binding sites only for androgens but not for estrogens suggesting a possible limitation to functional androgen receptors in this species. Further studies have to be conducted to shed more light into the discussion about the controversy about sex steroid receptors in invertebrates.

Amino Acid Sequence↗

An ecdysteroid-responsive gene in a lobster - a potential crustacean member of the steroid hormone receptor superfamily.

The role of ecdysteroids in modulating exoskeletal growth during the moult cycle of Crustacea has been well described. However, little is known about the action of ecdysteroids at the level of gene transcription and regulation in Crustacea. This paper reports the cloning of an ecdysteroid responsive gene, HHR3, a potential Manduca sexta MHR3 homologue in the American lobster, Homarus americanus. Levels of HHR3 expression are up-regulated in response to in vivo injections of premoult concentrations (10(-6) M) of 20-hydroxyecdysone in the epidermal and muscle tissue of the lobster after 6 h. Maximal mRNA levels are observed after 21 h before returning to basal levels. In muscle tissue, elevated levels of HHR3 mRNA follow a time course similar to elevated actin mRNA expression in response to hormonal injection. In contrast, in eyestalk tissue, the HHR3 levels decline up to 21 h post-injection before rising to basal levels after 48 h. Eyestalk, epidermal and leg muscle tissue was extracted over the moult cycle to determine the levels of expression. In muscle, HHR3 is high during the premoult period that corresponds to the period of the moult cycle when the ecdysteroid titre is high. In the epidermis, HHR3 levels are also high during the premoult with elevated levels maintained into the postmoult period. In the eyestalk, mRNA levels of HHR3 show an opposite pattern of expression with low levels during premoult and postmoult and high levels found during the intermoult period. Our results provide novel evidence for an ecdysteroid responsive gene in a crustacean that has many similarities to MHR3 in Manduca and DHR3 in Drosophila melanogaster. This raises the question of whether a similar cascade of ecdysteroid responsive genes exist in other members of Arthropoda such as the Crustacea, as has been demonstrated in Drosophila. In addition, we provide further evidence for negative feedback regulation of ecdysteroids at the site of moult-inhibiting hormone (MIH) production in the lobster eyestalk.

Amino Acid Sequence↗

Genome and EST analyses and expression of a gene family with putative functions in insect chemoreception.

Odorant-binding proteins (OBPs) are thought to be responsible for the transport of semiochemicals across hydrophobic interfaces to olfactory receptors. In insects, a second class of OBPs with four conserved cysteines has been variously named as sensory appendage proteins, olfactory segment-D proteins, and chemosensory proteins (CSPs). The physiological functions of these proteins have remained elusive. Here we report a comprehensive survey of both genome and expressed sequence tags (EST) databases. This showed that CSPs are apparently only present in the phylum, Arthropoda, and in two subphyla, Crustacea and Uniramia. This is the first report of a putative CSP in Crustacea and suggests that the origin of these genes predates the divergence of Uniramia and Crustacea. For the Uniramia, we identified 74 new genes encoding putative CSPs of insect species from 10 different orders. Using tissue-specific EST libraries, we have examined the relative expression of putative CSP genes in many tissues from 22 insect species suggesting that the genes are expressed widely. One Drosophila CSPs is expressed sixfold higher in head than other CSPs. One Bombyx mori CSPs was found at a very high level in pheromone gland, and for the first time, six CSPs were identified in B. mori compound eyes. The different frequencies of CSP transcripts were observed between solitary and gregarious EST libraries of Locusta migratoria.

Animals↗

[A case of tongueworms (Pentastomida): a specific problem in context of the modern phylogenetics].

A short essay on anatomy, ultrastructure and larval development, life cycles, classification, palaeontology and phylogenetic relationships of Pentastomida is given. Currently, the Pentastomida are usually placed within Crustacea, as sister group of Branchiura (Crustacea, Maxillopoda). The grounds are striking similarities in ultrastructures of spermatozoa and congruence in the 18S rRNA nucleotide sequence. Both taxa are however sharply disparate in all other characters concerning morphology, embryology, life cycles and geological history. A direct introduction of Pentastomida in the system of Crustacea implies an unjustified inflation of the taxonomic diagnosis of the latter group; hence it is unacceptable. Two alternative hypotheses are suggested as tentative phylogenetic relationships to be tested. Each hypothesis infers however a very complex and unlikely evolutionary scenario. 1) Pentastomida and Branchiura are true sister groups as justified by coincidence in spermatozoan ultrastructure and sequence of ribosomal RNA. Since their divergence, the evolutions of pentastomids and branchiurans ran in different milieus, in different shapes and with different rates. To present time, the pentastomids lost nearly all characters of maxillopods and crustaceans as well as majority of anthropod features. 2) Pentastomida and Branchiura are not close related. The similarity in ultrastructural details of spermatozoa may be occasional or resulted from convergent evolution with unknown factors. Coincidence in nucleotide sequences or ribosomal RNA should be reexamined and tested with other pentastomid species. The second hypothesis seems to be more likely.

Animals↗

The phylogenetic significance of crustacean optic neuropils and chiasmata: a re-examination.

Recent molecular data challenge the traditional hypotheses of arthropod phylogeny founded on morphologic characters. In this discussion, the structure of the visual systems in Pterygota (Hexapoda) and Decapoda (Malacostraca, Crustacea) is an important argument. Although many components of their visual systems depict structural homology, differences exist between Pterygota/Decapoda on the one side and Branchiopoda (Entomostraca) on the other in that the latter do not have a third optic neuropil or optic chiasmata. Therefore, the goals of the current study were to explore whether the third optic neuropils in Pterygota and Decapoda are homologous, to examine the formation of the first two optic neuropils and the chiasmata in Crustacea, and to compare these processes with Pterygota. For this purpose, five species of entomostracan and malacostracan crustaceans were analyzed by examination of serial sections, fluorescence labeling with phallotoxins, and anti-histamine immunohistochemistry. We found that the chiasmata of Decapoda and Pterygota are characterized by striking similarities regarding both the level of individually identifiable classes of neurons and ontogenetic mechanisms, which are clearly different from those in Branchiopoda. Furthermore, the third optic neuropil of Decapoda and Pterygota, the lobula, shares an ontogenetic protocerebral origin and an innervation by corresponding sets of histamine-immunoreactive neurons, suggesting homology of the lobula in these two groups. In conclusion, the characteristics of the visual system are in conflict with the traditional classification of Arthropoda. Instead, they support a sister-group relationship of Hexapoda and Malacostraca, as suggested by some of the molecular studies.

Animals↗

Efferent controls in crustacean mechanoreceptors.

Since the 1960s it has been known that central neural networks can elaborate motor patterns in the absence of any sensory feedback. However, sensory and neuromodulatory inputs allow the animal to adapt the motor command to the actual mechanical configuration or changing needs. Many studies in invertebrates, particularly in crustacea, have described several mechanisms of sensory-motor integration and have shown that part of this integration was supported by the efferent control of the mechanosensory neurons themselves. In this article, we review the findings that support such an efferent control of mechanosensory neurons in crustacea. Various types of crustacean proprioceptors feeding information about joint movements and strains to central neural networks are considered, together with evidence of efferent controls exerted on their sensory neurons. These efferent controls comprise (1) the neurohormonal modulation of the coding properties of sensory neurons by bioamines and peptides; (2) the presynaptic inhibition of sensory neurons by GABA, glutamate and histamine; and (3) the long-term potentiation of sensory-motor synapses by glutamate. Several of these mechanisms can coexist on the same sensory neuron, and the functional significance of such multiple modulations is discussed.

Animals↗

Toxicity of Euphorbia milii latex and niclosamide to snails and nontarget aquatic species.

The toxicity of Euphorbia milii molluscicidal latex and niclosamide (NCL) to target snails (Biomphalaria glabrata and Biomphalaria tenagophila) and nontarget aquatic organisms is evaluated. Planorbidae snails were killed by very low concentrations of lyophilized latex (48-h LC(50), mg/L: B. glabrata, 0.12; B. tenagophila, 0.09; Helisoma duryi, 0.10). Latex was less toxic (48-h LC(50) or EC(50), mg/L) to oligochaeta (Tubifex tubifex, 0.31), planktonic crustacea (Daphnia similis, 0.38; C. dubia, 1.07; Artemia sp., 0.93), and fishes (Danio rerio, 0.96; Poecilia reticulata, 1. 39), and considerably less toxic to Ampullariidae snails (Pomacea sp. , 10.55) and frog tadpoles (Rana catesbeiana, 7.50). Latex (up to 100 mg/L) was not toxic to bacteria (P. putida and V. fischeri), algae (Selenastrum capricornutum and Chlorella vulgaris), and mosquito larvae (Anopheles albitarsis, Aedes aegypti, Aedes fluviatilis). NCL was very toxic (48-h LC(50) or EC(50), mg/L) to Planorbidae snails (B. glabrata, 0.15, B. tenagophila, 0.13; H. duryi, 0.10), T. tubifex (0.11), crustacea (D. similis, 0.19; Ceriodaphnia dubia, 0.47; Artemia sp. 0.18), fishes (D. rerio, 0.25; P. reticulata, 0.29), R. catesbeiana (0.16), and Pomacea sp. (0.76). NCL was toxic to bacteria, algae (96-h IC(50), mg/L: S. capricornutum, 0.34; C. vulgaris, 1.23) and slightly toxic to mosquito larvae. In conclusion, E. milii latex, as compared with the reference molluscicide niclosamide, presents a higher degree of selectivity toward snails which are intermediate hosts of Schistosoma trematodes.

Animals↗

Monoaminergic neurons in the nervous system of crustaceans.

Certain neurons in the nervous system of the malacostracan crustaceans give rise to a predominantly green and a sparse yellow fluorophore in the histochemical fluorescence method of Falck-Hillarp. The same applies to the whole of Crustacea. The green fluorophore is probably a catecholamine; the yellow to brown-yellow has not yet been identified. The biogenic amine responsible for the green fluorescence, besides being found in diffusely distributed fibres, also appears in distinct areas of fibre concentrations in the central nervous system. The protocerebrum of the malacostracans contains three areas: the central body and two areas in the top of the brain, one anterior and one posterior. The latter two are not recognized as separate areas in ordinary histological preparations. In addition, the optic neuropiles are fluorescent, some with a distinct stratification of the fluorophore. The deuto- and tritocerebrum and the ventral nerve cord also contain monoaminergic neurons. Of the brightly fluorescent areas in the whole of Crustacea, only the central body consistently exists in all species. The other areas of concentrated fluorescent neuropile are restricted to smaller taxonomic units and differ from each other.

Animals↗

Analysis of the primary sequence and secondary structure of the unusually long SSU rRNA of the soil bug, Armadillidium vulgare.

The complete nucleotide sequence of the SSU rRNA gene from the soil bug, Armadillidium vulgare (Crustacea, Isopoda), was determined. It is 3214 bp long, with a GC content of 56.3%. It is not only the longest SSU rRNA gene among Crustacea but also longer than any other SSU rRNA gene except that of the strepsipteran insect, Xenos vesparum (3316 bp). The unusually long sequence of this species is explained by the long sequences of variable regions V4 and V7, which make up more than half of the total length. RT-PCR analysis of these two regions showed that the long sequences also exist in the mature rRNA and sequence simplicity analysis revealed the presence of slippage motifs in these two regions. The putative secondary structure of the rRNA is typical for eukaryotes except for the length and shape variations of the V2, V4, V7, and V9 regions. Each of the V2, V4, and V7 regions was elongated, while the V9 region was shortened. In V2, two bulges, located between helix 8 and helix 9 and between helix 9 and helix 10, were elongated. In V4, stem E23-3 was dramatically expanded, with several small branched stems. In V7, stem 43 was branched and expanded. Comparisons with the unusually long SSU rRNAs of other organisms imply that the increase in total length of SSU rRNA is due mainly to expansion in the V4 and V7 regions.

Animals↗

Marine invertebrate cell cultures: new millennium trends.

This review analyzes activities in the field of marine invertebrate cell culture during the years 1999 to 2004 and compares the outcomes with those of the preceding decade (1988 to 1998). During the last 5 years, 90 reports of primary cell culture studies of marine organisms belonging to only 6 taxa (Porifera, Cnidaria, Crustacea, Mollusca, Echinodermata, and Urochordata) have been published. This figure represents a 2-fold increase in the annual number of publications over the decade 1988 to 1998. Three other trends distinguish the two reviewed periods. First, in recent years studies attempting to improve cell culture methodologies have decreased, while interest in applications of already existing methodologies has increased. This reflects the effects of short-term cultures in attracting new researchers and scientific disciplines to the field. Second, only 17.8% of the recent publications used long-term cultures, compared with 30.0% of the publications in the previous decade. Third, during recent years research in cell cultures has studied fewer model species more extensively (mainly, Botryllus schlosseri, Crassostrea, Mytilus, Penaeus, and Suberites domuncula), signifying a shift from previous investigations that had studied a more diverse range of organisms. From 1988 to 1998 the phylum Mollusca was the most studied taxon (34.4%), but recent years have seen more studies of Porifera and Crustacea (30.0% and 32.2% of publications) than of Mollusca (21.1%). Still, not even a single established cell line from any marine invertebrate has yet been made available. However, the use of new cellular, genomic, and proteomic tools may fundamentally change our strategy for the development of cell cultures from marine invertebrates.

Animals↗

Structure and stability of arthropodan hemocyanin Limulus polyphemus.

In the hemolymph of many arthropodan species, respiratory copper proteins of high molecular weight, termed hemocyanins (Hcs) are dissolved. In this communication, we report on the protein stability of different hemocyanin species (Crustacea and Chelicerata) using fluorescence spectroscopy. Five to seven major electrophoretically separable protein chains (structural subunits) were purified by fast protein liquid chromatography (FPLC) ion exchange chromatography from different hemocyanins with very high sequence homology of the active site regions binding copper ions (CuA and CuB), and especially the relative sequence positions of histidine (His) and tryptophan (Trp) residues of these protein segments are in all cases identical. The conformational stabilities of the native dodecameric aggregates and their isolated structural subunits towards various denaturants (pH and guanidine hydrochloride (Gdn.HCl)) indicate that the quaternary structure is stabilized by hydrophilic and polar forces, whereby both, the oxy- and apo-forms of the protein are considered. These two classes of Crustacea and Chelicerata Hcs have the similar Trp-fluorescence quantum yields, but different values of lambda(max) emission (about 325 and 337 nm, respectively). Differences in the quantum yields are observed of the oxy- and apo-forms, which must be attributed to the fluorescence quenching effect of the two copper ions (CuA and CuB) in the active site. The position of emission maximum indicates tryptophan side chains are situated in a non-polar environment. Denaturation studies of Hcs by Gdn.HCl indicate that the denaturation process consists of two steps: dissociation of the native molecule into its structural subunits and denaturation of the subunits at concentrations >1.5M Gdn.HCl. Two steps of denaturation are also observed after keeping the protein in buffer solutions at different pH values with different pH-stability for holo-oxy and apo-Hc forms.

Amino Acid Sequence↗

Species sympatry and horizontal transfers of Mariner transposons in marine crustacean genomes.

Mariner-like elements (MLEs) have been widely detected in terrestrial species. The first complete MLE isolated from a marine invertebrate was detected in the genome of the hydrothermal crab Bythograea thermydron by Halaimia-Toumi et al. [Halaimia-Toumi, N., Casse, N., Demattei, M.V., Renault, S., Pradier, E., Bigot, Y., Laulier, M., 2004. The GC-rich transposon Bytmar1 from the deep-sea hydrothermal crab, Bythograea thermydron, may encode three transposase isoforms from a single ORF. J. Mol. Evol. 59, 747-760] and called Bytmar1. Here, we report the isolation of three new Bytmar1 relatives from the genomes of one hydrothermal amphipod Ventiella sulfuris (Vensmar1) and two coastal crustacea, Maia brachydactila (Maibmar1) and Cancer pagurus (Canpmar1). Like Bytmar1, these MLEs have an unusually high GC content, a high CpG ratio, and a low TpA ratio. Their consensus sequence encodes a transposase that is preceded by an N-flag, as in Bytmar1, which could be a marine feature. Only one of the 19 clones obtained, Vensmar1.3, encoded for a full-length transposase. The phylogenetic analyses revealed that all these Bytmar1-related elements can be differentiated into two clusters, corresponding to the coastal or hydrothermal origin of their hosts. They also confirmed that the irritans sub-family comprises at least four lineages that seem to depend on the taxonomical position and habitat of their hosts. Finally, we observed that elements coding for two potentially complete transposases exhibiting 99.5% similarity, Bytmar1.11 and Vensmar1.3, were present in the genome of two distantly related hydrothermal crustacea, one Amphipod and one Decapod. The hypothesis of horizontal transfers is discussed in the light of the sequence similarities observed.

Amino Acid Sequence↗

Crustacean ecdysteriods in reproduction and embryogenesis.

Ecdysteroids are the molting hormones in Crustacea, as in other arthropods. They also subserve functions in the control of reproduction and embryogenesis. The available evidence indicate that the ecdysteroids are sequestered into the ovary by binding to yolk precursor proteins. Steroidogenic ability of the ovary is yet to be demonstrated in Crustacea. Despite several investigations, the role of ecdysteroids in oocyte maturation is not fully known. However, the embryonic ecdysteroids undergo significant fluctuation, correlated to specific developmental stages, including the secretion of embryonic envelopes and cuticle. Ecdysteroid metabolism in the eggs seems to be active throughout embryogenesis inasmuch as the free ecdysteroids are rapidly converted into conjugates, and vice versa; in addition to their inactivation into excretory ecdysteroidic acids. Eyestalk neuropeptides such as molt inhibiting hormones have a dominant role on the ecdysteroid synthesis by Y-organ, although recent evidence suggests a stimulatory role for yet another endocrine gland, the mandibular organ on Y-organ synthesis.

Animals↗

Neurogenesis in the crustacean ventral nerve cord: homology of neuronal stem cells in Malacostraca and Branchiopoda?

In Insecta and malacostracan Crustacea, neurons in the ventral ganglia are generated by the unequal division of neuronal stem cells, the neuroblasts (Nbs), which are arranged in a stereotyped, grid-like pattern. In malacostracans, however, Nbs originate from ectoteloblasts by an invariant lineage, whereas Nbs in insects differentiate without a defined lineage by cell-to-cell interactions within the neuroectoderm. As the ventral ganglia in entomostracan crustaceans were thought to be generated by a general inward proliferation of ectodermal cells, the question arose as to whether neuroblasts in Euarthropoda represent a homologous type of stem cell. In the current project, neurogenesis in metanauplii of the entomostracan crustaceans Triops cancriformis Fabricius, 1780 (Branchiopoda, Phyllopoda) and Artemia salina Linné, 1758 (Branchiopoda, Anostraca) was examined by in vivo incorporation of the mitosis marker bromodeoxyuridine (BrdU) and compared to stem cell proliferation in embryos of the malacostracan Palaemonetes argentinus Nobili, 1901 (Eucarida, Decapoda). The developmental expression of synaptic proteins (synapsins) was studied immunohistochemically. Results indicate that in the ventral neurogenic zone of Branchiopoda, neuronal stem cells with cellular characteristics of malacostracan neuroblasts are present. However, a pattern similar to the lineage-dependent, grid-like arrangement of the malacostracan neuroblasts was not found. Therefore, the homology of entomostracan and malacostracan neuronal stem cells remains uncertain. It is now well established that during arthropod development, identical and most likely homologous structures can emerge, although the initiating steps or the mode of generation of these structures are different. Recent evidence suggests that adult Entomostraca and Malacostraca share corresponding sets of neurons so that the present report provides an example that those homologous neurons may be generated via divergent developmental pathways. In this perspective, it remains difficult at this point to discuss the question of common patterns of stem cell proliferation with regard to the phylogeny and evolution of Atelocerata and Crustacea.

Animals↗

Variability of fish consumption within the 10 European countries participating in the European Investigation into Cancer and Nutrition (EPIC) study.

OBJECTIVE: To describe and compare the consumption of total fish (marine foods) and the fish sub-groups - white fish, fatty fish, very fatty fish, fish products and crustacea, in participants from the European Investigation into Cancer and Nutrition (EPIC) study. DESIGN: Cross-sectional analysis of dietary intake using a computerised standardised 24-hour recall interview. Crude means, means and standard errors adjusted by age, season and day of the week were calculated, stratified by centre and gender. SETTING: Twenty-seven redefined centres in the 10 European countries participating in the EPIC study. SUBJECTS: In total, 35 955 subjects (13 031 men and 22 924 women), aged 35-74 years, selected from the main EPIC cohort. RESULTS: A six- to sevenfold variation in total fish consumption exists in women and men, between the lowest consumption in Germany and the highest in Spain. Overall, white fish represented 49% and 45% of the intake of total fish in women and men, respectively, with the greatest consumption in centres in Spain and Greece and the least in the German and Dutch centres. Consumption of fatty fish reflected that of total fish. However, the greatest intake of very fatty fish was in the coastal areas of northern Europe (Denmark, Sweden and Norway) and in Germany. Consumption of fish products was greater in northern than in southern Europe, with white fish products predominating in centres in France, Italy, Spain, The Netherlands and Norway. Intake of roe and roe products was low. The highest consumption of crustacea was found in the French, Spanish and Italian centres. The number of fish types consumed was greater in southern than in northern Europe. The greatest variability in consumption by day of the week was found in the countries with the lowest fish intake. CONCLUSIONS: Throughout Europe, substantial geographic variation exists in total fish intake, fish sub-groups and the number of types consumed. Day-to-day variability in consumption is also high.

Adult↗

Molecular phylogeny of the major arthropod groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods.

A phylogeny of the arthropods was inferred from analyses of amino acid sequences derived from the nuclear genes encoding elongation factor-1 alpha and the largest subunit of RNA polymerase II using maximum-parsimony, neighbor-joining, and maximum-likelihood methods. Analyses of elongation factor-1 alpha from 17 arthropods and 4 outgroup taxa recovered many arthropod clades supported by previous morphological studies, including Diplopoda, Myriapoda, Insecta, Hexapoda, Branchiopoda (Crustacea), Araneae, Tetrapulmonata, Arachnida, Chelicerata, and Malacostraca (Crustacea). However, counter to previous studies, elongation factor-1 alpha placed Malacostraca as sister group to the other arthropods. Branchiopod crustaceans were found to be more closely related to hexapods and myriapods than to malacostracan crustaceans. Sequences for RNA polymerase II were obtained from 11 arthropod taxa and were analyzed separately and in combination with elongation factor-1 alpha. Results from these analyses were concordant with those derived from elongation factor-1 alpha alone and provided support for a Hexapoda/Branchiopoda clade, thus arguing against the monophyly of the traditionally defined Atelocerata (Hexapoda + Myriapoda).

Amino Acid Sequence↗