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The complete mitochondrial genome of the orbiniid polychaete Orbinia latreillii (Annelida, Orbiniidae)--A novel gene order for Annelida and implications for annelid phylogeny.

Relationships of annelid subtaxa are controversially discussed and additional markers are necessarily needed to get further insights into their evolution. Due to their high content of information, mitochondrial genomes have been proven very useful in phylogenetic analyses. Whereas many complete mitochondrial genomes of arthropods are available, lophotrochozoan taxa are only scarcely represented and this is especially true for annelids. Here we present the complete mitochondrial genome of the orbiniid polychaete Orbinia latreillii. The circular genome is 15,558 bp in size and contains the same 37 genes as found in most other metazoans. As in the case for all studied annelids all genes are transcribed from the same strand. Compared with the known data from other annelids at least five gene translocations must be hypothesized for O. latreillii. A comparison of the available data shows that gene translocations within Annelida seem to be less frequent than in molluscs, but more frequent as previously assumed. Phylogenetic analyses of mitochondrial DNA sequence data and amino acid data support an inclusion of Sipuncula within Annelida and a closer relationship to orbiniids is recovered for this taxon.

Animals↗

Comparison of kinetic constants of creatine kinase isoforms.

The purpose of this study was to elucidate the functional differences between the CK isoforms by cloning the cDNAs of 12 CK isoforms: the M and B cytoplasmic forms and uMiCK from mouse, the M1, M2 and B cytoplasmic forms from Danio rerio, M1 and M2 cytoplasmic forms from the lower vertebrate Lampetra japonica, a cytoplasmic CK and a MiCK from the marine worm Neanthes diversicolor, and a cytoplasmic CK and a MiCK from the soft coral Dendronephthya gigantea. These were expressed in Escherichia coli as a fusion protein with maltose-binding protein, and kinetic constants (K(m), K(d) and k(cat)) of all the recombinant enzymes, except for the unstable Dendronephthya cytoplasmic CK, were determined for the forward reaction. The kinetic constants of the M- and B-forms of the mouse and Danio cytoplasmic CKs differed significantly, with the K(m) for creatine (K(m)Cr) of M-CK being three- to nine-fold higher than that of B-CK, possibly reflecting differences in the concentration of creatine in muscle and brain cells. The mouse uMiCK had the lowest K(m)Cr value among the CK isoforms. In addition, it also exhibited a strong synergism for substrate binding (K(d)/K(m)=11.8). These results indicate that uMiCK has unique characteristics compared with other CK isoforms. Two subisoforms of M-CK were found in the lower vertebrate L. japonica, and the kinetic constants of recombinant M1- and M2-CKs differed significantly. The M1- and M2-CKs were expressed in skeletal muscle with a ratio of 7:3, while M1-CK was the predominant subisoform in the testis. The kinetic constants of cytoplasmic CK from the marine worm Neanthes were significantly different from those of Neanthes MiCK, possibly indicating that functional differences among CK isoforms occurred at least before the divergence of annelids from other protostome invertebrates.

Animals↗

Spectroscopic characterization of the ferric states of Amphitrite ornata dehaloperoxidase and Notomastus lobatus chloroperoxidase: His-ligated peroxidases with globin-like proximal and distal properties.

Amphitrite ornata dehaloperoxidase (DHP) and Notomastus lobatus chloroperoxidase (NCPO) catalyze the peroxide-dependent dehalogenation of halophenols and halogenation of phenols, respectively. Both enzymes have histidine (His) as their proximal heme iron ligand. Crystallographic examination of DHP revealed that it has a globin fold [M.W. LaCount, E. Zhang, Y.-P. Chen, K. Han, M.M. Whitton, D.E. Lincoln, S.A. Woodin, L. Lebioda, J. Biol. Chem. 275 (2000) 18712-18716] and kinetics studies established that ferric DHP is the active state [R.L. Osborne, L.O. Taylor, K. Han, B. Ely, J.H. Dawson, Biochem. Biophys. Res. Commun. 324 (2004) 1194-1198]. NCPO likely has these same properties. Previous work with His-ligated heme proteins has revealed characteristic spectral distinctions between dioxygen binding globins and peroxide-activating peroxidases. Since DHP, and likely NCPO, is a peroxide-activating globin, we have sought to determine in the present investigation whether the ferric resting states of these two novel heme-containing enzymes are myoglobin-like or peroxidase-like. To do so, we have examined their exogenous ligand-free ferric states as well as their azide, imidazole and NO bound ferric adducts (and ferrous-NO complexes) with UV-Visible absorption and magnetic circular dichroism spectroscopy. We have also compared each derivative to the analogous states of horse heart myoglobin (Mb) and horseradish peroxidase (HRP). The spectra observed for parallel forms of DHP and NCPO are virtually identical to each other as well as to the spectra of the same Mb states, while being less similar to the spectra of corresponding HRP derivatives. From these data, we conclude that exogenous ligand-free ferric DHP and NCPO are six-coordinate with water and neutral His as ligands. This coordination structure is distinctly different from the ferric resting state of His-ligated peroxidases and indicates that DHP and NCPO do not activate bound peroxide through a mechanism dependent on a push effect imparted by a partially ionized proximal His as proposed for typical heme peroxidases.

Animals↗

Low resolution crystal structure of Arenicola erythrocruorin: influence of coiled coils on the architecture of a megadalton respiratory protein.

Annelid erythrocruorins are extracellular respiratory complexes assembled from 180 subunits into hexagonal bilayers. Cryo-electron microscopic experiments have identified two different architectural classes. In one, designated type I, the vertices of the two hexagonal layers are partially staggered, with one hexagonal layer rotated by about 16 degrees relative to the other layer, whereas in the other class, termed type II, the vertices are essentially eclipsed. We report here the first crystal structure of a type II erythrocruorin, that from Arenicola marina, at 6.2 A resolution. The structure reveals the presence of long continuous triple-stranded coiled-coil "spokes" projecting towards the molecular center from each one-twelfth unit; interdigitation of these spokes provides the only contacts between the two hexagonal layers of the complex. This arrangement contrasts with that of a type I erythrocruorin from Lumbricus terrestris in which the spokes are broken into two triple-stranded coiled coils with a disjointed connection. The disjointed connection allows formation of a more compact structure in the type I architecture, with the two hexagonal layers closer together and additional extensive contacts between the layers. Comparison of sequences of the coiled-coil regions of various linker subunits shows that the linker subunits from type II erythrocruorins possess continuous heptad repeats, whereas a sequence gap places these repeats out of register in the type I linker subunits, consistent with a disjointed coiled-coil arrangement.

Amino Acid Sequence↗

A public software for energy filtering transmission electron tomography (EFTET-J): application to the study of granular inclusions in bacteria from Riftia pachyptila.

Energy-filtering transmission electron microscopy (EFTEM) allows the determination of elemental distributions out of a sequence of energy filtered images. Combined with electron tomography, EFTEM is a powerful tool to obtain three-dimensional chemical maps from sub-cellular structures. However, there is no existing software in the public-domain for the computation and analysis of 3D-chemical maps. Here, we present a Java-based program to compute 3D-elemental distribution. This program is available as a set of plug-ins for the public-domain Java image processing program Image J inspired by NIH Image. Its implemented algorithms have been successfully applied to the three-dimensional localization of iron granules in semi thin (200 nm) epon sections from the vent worm Riftia pachyptalia.

Algorithms↗

pH effects on the binding of oxygen to non-vertebrate monomeric hemoglobins. A linked function model.

Monomeric invertabrate hemoglobins with high oxygen affinity usually contain a tyrosine in the distal region of the heme. This feature has stimulated investigations revealing that one of the properties resulting from the presence of the distal tyrosines is a decreased off rate on the binding of oxygen, thus developing the high affinity. Despite that fact that the pK value of the tyrosine group differs significantly from the groups it replaces little attention has been paid to the pH dependence of the binding of oxygen to the high affinity hemoglobins. Such a pH dependence has been reported on two of the monomeric hemoglobins with relatively low oxygen affinity and one monomeric hemoglobin of intermediate affinity. The pH data of these hemoglobins has been analysed with a linked function model involving the hydrogen ion. pK values required for the low-affinity hemoglobins vary from 4.5 to 7.5. When applied to the high-affinity hemoglobins, the linked function model provides reasonable values for the binding parameters. These pK values vary from 3.0 to 9.0.

Animals↗

The relative importance of water and diet for uptake and subcellular distribution of cadmium in the deposit-feeding polychaete, Capitella sp. I.

The impact of dietary and water exposure on the accumulation and distribution of cadmium (Cd) in subcellular components of the polychaete Capitella sp. I was investigated. Worms were exposed to either dissolved Cd alone ('Water-Only' treatments; WO) or diet-bound Cd alone ('Algae-bound Only' treatments; AO). Thus, WO worms were starved and AO worms were fed. Differential centrifugation was used to fractionate worm homogenates into debris- (DE), mitochondrial- (MI), microsomal- (MC) and cytosolic- (CY) fractions, and the concentration of Cd in these fractions was quantified by radiometric analysis using the specific activity of the (109)Cd radioisotope. Neither exposure route nor Cd exposure affected the total protein content in the worms. However, fed, but not starved worms relocated proteins from the mitochondria to the cytosol in response to Cd exposure. In systems without Cd addition, starvation likewise influenced the distribution of protein between mitochondria and cytosol. Cutaneous uptake and accumulation of Cd from the water was related to surface area while dietary uptake was influenced by the amount of sediment passing through the gut. Irrespective of exposure route, Cd was accumulated primarily in the debris fraction although comparisons between the treatments indicated that relatively less Cd was located in DE and more in MI, MC and CY fractions in worms in AO compared to worms in WO. Cd per unit protein in DE and MI fractions was twice as high as in MC and CY fractions in WO, whereas MC contained the highest Cd content followed by MI, DE and CY fractions in AO worms. Our results stress the importance of exposure route for the internal distribution and toxicity of Cd to deposit feeders such as Capitella sp. I.

Analysis of Variance↗

Biomarker responses to pollution in two invertebrate species: Scrobicularia plana and Nereis diversicolor from the Cádiz bay (SW Spain).

The clam Scrobicularia plana and the polychaete worm Nereis diversicolor were collected in several sites from a littoral enclosure in SW Spain. The aim of our study was to relate various biomarker responses in these species to a pollution gradient caused by untreated domestic discharges and to verify the adequacy of the selected species as sentinels in this habitat. The biomarkers selected were the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX) and DT-diaphorase (DT-D). In addition, the activities of cytochrome P450-dependent ethoxyresorufin O-deethylase (EROD) activity, the phase II detoxifying enzyme glutathione S-transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE) were measured. Metallothionein levels were selected as biomarkers of heavy metals exposure in both species. The results suggest a different response in the water filtering organism (clam) and the sediment eater (polychaete), probably as a consequent of different pollution exposure and that samples from the "Caño Sancti-Petri" were exposed to biologically active compounds that altered some of their biochemical responses. AChE was the most sensitive biomarker in both species and N. diversicolor proved to be a more robust sentinel in this ecosystem.

Acetylcholinesterase↗

Total content and chemical speciation of arsenic in the polychaete Sabella spallanzanii.

An unusual feature has been observed in polychaetes regarding their capacity to accumulate high levels of relatively toxic forms of arsenic in specific tissues. Basal levels of arsenic and distribution of its compounds were investigated in tissues of the Mediterranean polychaete Sabella spallanzanii. Particularly high concentrations were measured in the branchial crown (1036+/-136 microg/g d.w.) and chemical speciation revealed that the predominant form was the relatively toxic dimethylarsinate (DMA). These data suggest a potential role of As as anti-predatory strategy in polychaetes.

Animals↗

The immediate effects of Hurricane Iniki on intertidal fauna on the south shore of O'ahu.

When Hurricane Iniki struck the Hawaiian Islands in September 1992, it provided a rare opportunity to examine the immediate effects of a hurricane on two intertidal benthic communities off the reefs of O'ahu, Hawai'i. The Niu Beach site contained large, obvious aggregations of the tube building polychaete Diopatra dexiognatha, and the Wailupe Beach site was without obvious tubiculous fauna at the surface. Ten replicate sediment cores were taken before and after the hurricane with a 7.6 cm PVC corer and organisms were identified to family and enumerated. There were no substantial depletions or loss of taxa after the hurricane. Oligochaetes were the most dominant taxa pre-and post-hurricane. The abundance of all dominant polychaete families increased post-hurricane. The three most abundant polychaetes were capitellids and D. dexiognatha (Onuphidae) at Niu Beach and Pygospio muscularis (Spionidae) at Wailupe Beach. We suggest that D. dexiognatha and P. muscularis help stabilize the sediments since they both form dense tube mats while capitellids and oligochaetes are considered highly adaptive surface burrowers that can take advantage of newly disturbed sediments. Overall, there was no substantial effect observed on the intertidal fauna exposed to this severe disturbance. It is suggested here that invertebrate communities in this area are adapted to survive and thrive in high-energy environments and possibly benefit from dense aggregations of tube building polychaetes.

Animals↗

Exposure to 2,4-decadienal negatively impacts upon marine invertebrate larval fitness.

Diatoms liberate volatile, biologically active unsaturated aldehydes following cell damage, which negatively impact upon invertebrate reproductive processes such as fertilization, embryogenesis and larval survival. 2,4-Decadienal is frequently identified among the aldehydes produced and is one of the more biologically active. The majority of studies which have examined the toxic effects of diatom aldehydes to invertebrate reproduction have scored egg production and/or hatching success as indicators of biological impacts. There are very few studies which have dealt specifically with the impacts of diatom-derived aldehydes on larval fitness. Larval stages of the polychaetes Arenicola marina and Nereis virens and the echinoderms Asterias rubens and Psammechinus miliaris exposed to 2,4-decadienal at sub 1 microg ml(-1) concentrations suffered reduced survival over the incubation period (day 1-8 post fertilization) with detectable differences for the polychates at a concentration of 0.005 and 0.01-0.1 microg ml(-1) for the echinoderms. Susceptibility of larval N. virens was investigated using stage specific 24 h exposures at 2,4-decadienal concentrations up to 1.5 microg ml(-1). A clear stage specific effect was found, with earlier larval stages most vulnerable. Nectochaete larvae (9-10 d) showed no reduction in survival at the concentrations assayed. Fluctuating asymmetry (FA), defined as random deviations from perfect bilateral symmetry, was used to analyse fitness of larval P. miliaris exposed to 2,4-decadienal at concentrations of 0.1, 0.5 and 1 microg ml(-1). The degree and frequency of asymmetrical development increased with increasing 2,4-decadienal concentration. Equally, as FA increased larval survival decreased. These results provide further support for the teratogenic nature of 2,4-decadienal and its negative impact on invertebrate larval fitness.

Aldehydes↗

Extending the generality of ecological models to artificial floating habitats.

Marine assemblages on natural hard substrata are generally different from those on artificial habitats. There is, however, the potential for certain ecological processes to operate on both types of structures. On the sides of floating pontoons in Sydney Harbour, there were strong patterns of vertical distribution of sessile epibiotic organisms and molluscan grazers across relatively small spatial scales (in three defined zones, namely splash, shallow and deep). Patterns of vertical distribution of the tubeworms Hydroides spp. were reversed depending on the cover of mussels. A manipulative experiment was done to test if patterns of vertical distribution of Hydroides spp. were due to (1) the functioning of mussels or (2) the structure provided by mussels. Neither the functioning nor structure of mussels accounted for the patterns of distribution of Hydroides spp. Mussels increased recruitment of Hydroides spp., in the shallow and deep zones, and this was not due to increased surface area of the mussel shells. Manipulation of numbers of grazers and covers of sessile epibiota showed that the observed negative relationship between grazers and epibiota was due to grazers reducing recruitment of epibiota and epibiota decreasing survival of grazers. Most importantly, processes that accounted for patterns of distribution of mobile and sessile organisms on artificial floating structures were similar to those repeatedly shown to create such patterns on natural rocky shores.

Animals↗

Sabella spallanzanii filter-feeding on bacterial community: ecological implications and applications.

The filtration process of Sabella spallanzanii Gmelin on bacterial community was studied in a coastal area of the Northern Ionian Sea (Mediterranean Sea) at three sites, S. Vito, Lido Gandoli and Lido Silvana, where some specimens of S. spallanzanii were transplanted. Analyses were performed both on water and worm samples. A total of six microbial groups were examined: culturable heterotrophic bacteria, total culturable bacteria at 37 degrees C, culturable vibrios, total and fecal coliforms and fecal streptococci. The bacterial densities were usually orders of magnitude higher in the worm homogenates than in the corresponding seawater and the highest values were observed in August. The ability of S. spallanzanii to accumulate the microbial pollution indicators suggests this species can be employed as a bioindicator for monitoring water quality. Moreover, the accumulation capability of S. spallanzanii for specific micro-organisms provides a potential role in sewage bioremediation.

Analysis of Variance↗

Bioaccumulation of PCB congeners in marine benthic infauna.

Field concentrations of polychlorinated biphenyls (PCBs) were measured in sediment and nine marine soft bottom invertebrate species. Lipid- and organic carbon normalised biota-sediment accumulation factors (BSAFs) were determined for 29 nonplanar and 11 coplanar congeners. To investigate whether the bioaccumulation was in thermodynamic equilibrium with the sediment, the determined BSAFs were compared to theoretically calculated ones to obtain a BSAFdet./BSAFtheor-ratio. Large interspecific variations were found: one suspension feeding and one deposit feeding species of brittle stars (Amphiura filiformis and A. chiajei), and one predatory polychaete (Glycera rouxii) had ratios>1 for congeners with logKow>6.5. In these species there was also a linear relationship between logKow and BSAF, both for coplanar and for planar congeners but with lower values for coplanar ones. For other species the pattern was more scattered. Only the deposit feeding polychaete Melinna cristata had BSAFdet./BSAFtheor-ratios<1 for all congeners. Thus, the interspecific variations in bioaccumulation did not correlate with differences in feeding strategies, but may be caused by differences in biotransformation, and in age and size of the analysed specimens.

Age Factors↗

Tracing the anthropogenic lead sources in coastal sediments of SE-pacific (36 degrees Lat. S) using stable lead isotopes.

This study evaluates the main sources of antropogenic Pb in one of the most industrialized centers of the southern Chilean coast (36 degrees S). Stable lead isotopes ((206)Pb/(207)Pb, (208)Pb/(207)Pb) were used to trace main Pb sources to coastal sediments, considering the suspended particulate matter (SPM) from marine (traps), continental (rivers) and industrial effluents, sediments and leaded gasoline samples. The atmospheric input was evaluated through natural collectors; i.e. Raqui-Tubul salt marsh. Results show that marine samples lie on a trend between industrial effluents ( approximately 1.16, 2.44) and natural sources (1.20, 2.50), not related to gasoline consumption. Salt marsh sediments show comparable isotopic composition to marine samples, suggesting the importance of the atmospheric input in the coastal sediments, not related to the leaded gasoline composition either. The continental input (1.18, 2.48) is highly influenced by precipitation, being difficult to separate both sources (atmosphere and continental runoff), showing also similar isotopic ratio to marine sediments. The signal of industrial emissions is masked with the introduction of Pb with higher isotopic ratios, compared to the values observed in the material collected from traps (SPM approximately 1.19, 2.48). The contribution of more radiogenic Pb by the upwelling is suggested.

Air Pollutants↗

Benthic changes during 10 years of organic enrichment by McMurdo Station, Antarctica.

A benthic habitat along the coast of McMurdo Station in the Ross Sea, Antarctica is enriched by sewage from the station and altered by hydrocarbons and heavy metals in an adjacent historic dumpsite. We report on 10 years of change in the benthic communities from 1988 to 1998 and compare enrichment effects at Australia's Casey Station, East Antarctica. Despite being 14 km apart, reference communities upcurrent and downcurrent of McMurdo Station remained closely similar over time, dominated in all years by a tube building polychaete, Spiophanes tcherniae. The community bordering McMurdo Station was generally a third as abundant as communities at the reference sites over the decade of sampling, although diversity was as high or higher, except in the most contaminated areas. In 1992, organic enrichment of the outfall community intensified and within the year, the opportunistic polychaetes Aphelochaeta sp., Ophryotrocha notialis, Capitella perarmata, and Leitoscoloplos kerguelensis became dominant. Since 1996, two of the three enriched communities have increased in resemblance to the reference communities. Given the observed responsiveness of the benthos to the outfall so far, further changes are anticipated within the year following implementation of sewage treatment in 2003. Organic enrichment by McMurdo Station has had a greater impact on benthic community structure than at Australia's Casey Station.

Animals↗

Polychaetes as environmental indicators revisited.

The utilization of polychaetes in descriptive ecology is reviewed in the light of recent research especially concerning the biota hard bottom environments. Polychaetes, often linked in the past to the concept of opportunistic species able to proliferate after an increase in organic matter, have played an important role especially with regard to impacted soft-bottom habitats. Increased knowledge of the group, suggests that not only opportunistic species can be utilised as indicators, so that these organisms can be disengaged from the old concept of opportunistic taxa. Moreover, recent researches conducted on this group allowed demonstrating as surrogacy is not always applicable. Among polychaetes inhabiting hard bottom environment, the analysis of family Syllidae appears particularly promising. Studied conducted in our laboratory demonstrated as syllid species decrease in abundance or completely disappear under varying sources of negative impact. The distribution of species also appeared indicative in underlying effects of marine protected areas (MPA) functioning, or in describing different climatic areas within biogeographical sectors. It is obvious that good results can only be obtained on the basis of good taxonomic resolution. We suggested that, in monitoring studies, operational time could be optimized not only by working at a higher-level on the whole invertebrate data set, but by also selecting a particularly indicative group and working at fine level.

Animals↗