Sea-floor videos and photographs (benthos) along 6 shelf profiles from the eastern Weddell Sea, Antarctica taken with remote operated vehicel CHEROKEE during Polarstern cruise ANT-XXI/2

The ROV operations had three objectives: (1) to check, whether the "Cherokee" system is suited for advanced benthological work in the high latitude Antarctic shelf areas; (2) to support the disturbance experiment, providing immediate visual Information; (3) to continue ecological work that started in 1989 at the hilltop situated at the northern margin of the Norsel Bank off the 4-Seasons Inlet (Weddell Sea). The "Cherokee" is was equipped with 3 video cameras, 2 of which support the operation. A high resolution Tritech Typhoon camera is used for scientific observations to be recorded. In addition, the ROV has a manipulator, a still camera, lights and strobe, compass, 2 lasers, a Posidonia transponder and an obstacle avoidance Sonar. The size of the vehicle is 160 X 90 X 90cm. In the present configuration without TMS (tether management system) the deployment has to start with paying out the full cable length, lay it in loops on deck and connect the glass fibres at the tether's spool winch. After a final technical check the vehicle is deployed into the water, actively driven perpendicular to the ship's axis and floatings are fixed to the tether. At a cable length of approx. 50 m, the tether is tightened to the depressor by several cable ties and both components are lowered towards the sea floor, the vehicle by the thruster's propulsion and the depressor by the ship's winch. At 5 m intervals the tether has to be tied to the single conductor cable. In good weather conditions the instruments supporting the navigation of the ROV, especially the Posidonia system, allow an operation mode to follow the ship's course if the ship's speed is slow. Together with the lasers which act as a scale in the images they also allow a reproducible scientific analysis since the transect can be plotted in a GIS system. Consequently, the area observed can be easily calculated. An operation as a predominantly drifting system, especially in areas with bottom near currents, is also possible, however, the connection of the tether at the rear of the vehicle is unsuitable for such conditions. The recovery of the system corresponds to that of the deployment. Most important is to reach the surface of the sea at a safe distance perpendicular to the ship's axis in order not to interfere with the ship's propellers. During this phase the Posidonia transponder system is of high relevance although it has to be switched off at a water depth of approx. 40 m. The minimum personal needed is 4 persons to handle the tether on deck, one person to operate the ship's winch, one pilot and one additional technician for the ROV's operation itself, one scientist, and one person on the ship's bridge in addition to one on deck for whale watching when the Posidonia system is in use. The time for the deployment of the ROV until it reaches the sea floor depends on the water depth and consequently on the length of the cable to be paid out beforehand and to be tightened to the single conductor cable. Deployment and recovery at intermediate water depths can last up to 2 hours each. A reasonable time for benthological observations close to the sea floor is 1 to 3 hours but can be extended if scientifically justified. Preliminary results: after a first test station, the ROV was deployed 3 times for observations related to the disturbance experiment. A first attempt to Cross the hilltop at the northern margin of the Norsel Bank close to the 4- Seasons Inlet was successful only for the first hundreds of metres transect length. The benthic community was dominated in biomass by the demosponge Cinachyra barbata. Due to the strong current of approx. 1 nm/h, the design of the system, and an expected more difficult current regime between grounded icebergs and the top of the hilltop the operation was stopped before the hilltop was reached. In a second attempt the hilltop was successfully crossed because the current and wind situation was much more suitable. In contrast to earlier expeditions with the "sprint" ROV it was the first time that both slopes, the smoother in the northeast and the steeper in the southwest were continuously observed during one cast. A coarse classification of the hilltop fauna shows patches dominated by single taxa: cnidarians, hydrozoans, holothurians, sea urchins and stalked sponges. Approximately 20 % of the north-eastern slope was devastated by grounding icebergs. Here the sediments consisted of large boulders, gravel or blocks of finer sediment looking like an irregularly ploughed field. On the Norsel Bank the Cinachyra concentrations were locally associated with high abundances of sea anemones. Total observation time amounted to 11.5 hours corresponding to almost 6-9 km transect length.

Two mitochondrial group I introns in a metazoan, the sea anemone Metridium senile: one intron contains genes for subunits 1 and 3 of NADH dehydrogenase.

Mitochondrial genes for cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 5 (ND5) of the sea anemone Metridium senile (phylum Cnidaria) each contain a group I intron. This is in contrast to the reported absence of introns in all other metazoan mtDNAs so far examined. The ND5 intron is unusual in that it ends with A and contains two genes (ND1 and ND3) encoding additional subunits of NADH dehydrogenase. Correctly excised ND5 introns are not circularized but are precisely cleaved near their 3' ends and polyadenylylated to provide bicistronic transcripts of ND1 and ND3. COI introns, which encode a putative homing endonuclease, circularize, but in a way that retains the entire genome-encoded intron sequence (other group I introns are circularized with loss of a short segment of the intron 5' end). Introns were detected in the COI and ND5 genes of other sea anemones, but not in the COI and ND5 genes of other cnidarians. This suggests that the sea anemone mitochondrial introns may have been acquired relatively recently.

Synergistic action of actinoporin isoforms from the same sea anemone species assembled into functionally active heteropores

Among the toxic polypeptides secreted in the venom of sea anemones, actinoporins are pore forming toxins whose toxic activity relies on the formation of oligomeric pores within biological membranes. Intriguingly, actinoporins appear as multigene families which give rise to many protein isoforms in the same individual displaying high sequence identities but large functional differences. However, the evolutionary advantage of producing such similar isotoxins is not fully understood. Here, using sticholysins I and II (StnI and StnII) from the sea anemone Stichodactyla helianthus, it is shown that actinoporin isoforms can potentiate each other’s activity. Through hemolysis and calcein releasing assays, it is revealed that mixtures of StnI and StnII are more lytic than equivalent preparations of the corresponding isolated isoforms. It is then proposed that this synergy is due to the assembly of heteropores since (i) StnI and StnII can be chemically cross-linked at the membrane and (ii) the affinity of sticholysin mixtures for the membrane is increased with respect to any of them acting in isolation, as revealed by isothermal titration calorimetry experiments. These results help to understand the multigene nature of actinoporins and may be extended to other families of toxins that require oligomerization to exert toxicity.

Synergistic Action of Actinoporin Isoforms from the Same Sea Anemone Species Assembled into Functionally Active Heteropores

Among the toxic polypeptides secreted in the venom of sea anemones, actinoporins are the pore-forming toxins whose toxic activity relies on the formation of oligomeric pores within biological membranes. Intriguingly, actinoporins appear as multigene families that give rise to many protein isoforms in the same individual displaying high sequence identities but large functional differences. However, the evolutionary advantage of producing such similar isotoxins is not fully understood. Here,using sticholysins I and II (StnI and StnII) from the sea anemone Stichodactyla helianthus, it is shown that actinoporin isoforms can potentiate each other’s activity. Through hemolysis and calcein releasing assays, it is revealed that mixtures of StnI and StnII are more lytic than equivalent preparations of the corresponding isolated isoforms. It is then proposed that this synergy is due to the assembly of heteropores because (i) StnI and StnII can be chemically cross-linked at the membrane and (ii) the affinity of sticholysin mixtures for the membrane is increased with respect to any of them acting in isolation, as revealed by isothermal titration calorimetry experiments. These results help us understand the multigene nature of actinoporins and may be extended to other families of toxins that require oligomerization to exert toxicity.

Purification and characterization of the biological effects of phospholipase A(2) from sea anemone Bunodosoma caissarum

Sea anemones contain a variety of biologically active substances. Bunodosoma caissarum is a sea anemone from the Cnidaria phylum, found only in Brazilian coastal waters. The aim of the present work was to study the biological effects of PLA(2) isolated from the sea anemone B. caissarum on the isolated perfused kidney, the arteriolar mesenteric bed and on insulin secretion. Specimens of B. caissarum were collected from the Sao Vicente Channel on the southern coast of the State of São Paulo, Brazil. Reverse phase HPLC analysis of the crude extract of B. caissarum detected three PLA(2) proteins (named BcPLA(2)1, BCPLA(2)2 and BcPLA(2)3) found to be active in B. caissarum extracts. MALDI-TOF mass spectrometry of BcPLA(2)1 showed one main peak at 14.7 kDa. The N-terminal amino acid sequence of BcPLA(2)1 showed high amino acid sequence identity with PLA(2) group III protein isolated from the Mexican lizard (PA23 HELSU, HELSU, PA22 HELSU) and with the honey bee Apis mellifera (PLA(2) and 1POC_A). In addition, BcPLA(2)1 also showed significant overall homology to bee PLA(2). The enzymatic activity induced by native BCPLA(2)1 (20 mu g/well) was reduced by chemical treatment with p-bromophenacyl bromide (p-BPB) and with morin. BcPLA(2)1 strongly induced insulin secretion in presence of high glucose concentration. In isolated kidney, the PLA(2) from B. caissarum increased the perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate, and sodium, potassium and chloride levels of excretion. BcPLA(2)1, however, did not increase the perfusion pressure on the mesenteric vascular bed. In conclusion, PLA(2), a group III phospholipase isolated from the sea anemone B. caissarum, exerted effects on renal function and induced insulin secretion in conditions of high glucose concentration. (C) 2009 Elsevier Ltd. All rights reserved.

Role of the Tryptophan Residues in the Specific interaction of the Sea Anemone Stichodacty la helianthus’s Actinoporin Sticholysin II with Biological Membranes

Actinoporins are pore-forming toxins from sea anemones. Upon interaction with sphingomyelin-containing bilayers, they become integral oligomeric membrane structures that form a pore. Sticholysin II from Stichodactyla helianthus contains five tryptophans located at strategic positions; its role has now been studied using different mutants. Results show that W43 and W115 play a eterminant role in maintaining the high thermostability of the protein, while W146 provides specific interactions for protomer−protomer assembly. W110 and W114 sustain the hydrophobic effect, which is one of the major driving forces for membrane binding in the presence of Chol. However, in its absence, additional interactions with sphingomyelin are required. These conclusions were confirmed with two sphingomyelin analogues, one of which had impaired hydrogen bonding properties. The results obtained support actinoporins’ Trp residues playing a major role in membrane recognition and binding, but their residues have an only minor influence on the diffusion and oligomerization steps needed to assemble a functional pore.

Marine flora and fauna of the Eastern United States Anthozoa: Acitniaria, Corallimorpharia, Ceriantharia, and Zoanthidea

This key includes 60 species of sea anemones and their relatives in the orders Actiniaria, Corallimorpharia, Ceriantharia, and Zoanthidea. Species from the intertidal zone, continental slope, and deep sea are included over a geographic range from Atlantic Canada to approximately South Carolina. In addition to the illustrated key itself, characteristics of each species are summarized in tabular form, including morphology, distribution, and types and sizes of cnidae. Ecological and taxonomic information on each species are also included in an annotated species list. (PDF file contains 76 pages.)

Biologische Veränderungen im Versenkungsgebiet für schwach radioaktive Abfälle im Iberischen Tiefseebecken?

From 1979 to 2000, the site for the dumping of low-level radioactive wastes and an unaffected site in the North-east Atlantic were investigated to determine the biomass data of the benthopelagic nekton and the benthic organisms. The investigation shall demonstrate the influence on the biocoenosis of the so-called "artificial reef effect", caused by dumped waste drums. For sampling benthopelagic and benthic organisms, we used a modified Agassiz trawl, called the Deep-sea Closing Net. With the exception of the “Amperima event” in 1996 – a unique occurrence of a large number of sea-cucumbers – there was no change in the composition of benthopelagic and benthic organisms through the years. The biomass of the benthopelagic nekton was characterized by Macrouridae(rat-tailed fishes) and the main tax on of the benthos was Actiniaria (sea-anemones).

The taxonomic position of the pelagic `staurozoan` Tessera gemmaria as a ceriantharian larva

Based on 16 specimens from the Southwestern Atlantic coast (Argentina and Brazil) we reinterpret the taxonomic position of Tessera gemmaria Goy, 1979, a stauromedusa considered as incertae sedis for a long time. Using external morphology, histological preparations and molecular data (16S and COI) we conclude that T. gemmaria is an early stage of a cerinula, the long-lived planktonic larval stage of the Ceriantharia (Anthozoa).

Biological features of a puzzling symbiotic association between the hermit crab Dardanus insignis and the porcellanid crab Porcellana Sayana (Crustacea)

The symbiotic lifestyle is widespread among porcellanid crabs, which maintain ecological and co-evolutionary associations with annelid polychaetes, poriferans, cnidarians, echinoderms, gastropod. mollusks, and other crustaceans such as shrimps and hermit crabs, among others. We investigated the ecological association between the hermit crab Dardanus insignis and the porcellanid Porcellana sayana, in southeastern Brazil. Porcellanid crabs, hermit crabs, and available shells were collected monthly from July 2001 to June 2003, with a shrimp boat equipped with two double-rig trawl nets. The majority of P. sayana specimens were collected in shells occupied by D. insignis (96.6%); a few were found in empty shells (3.4%). The catch of both symbionts and hosts increased with increasing depth, with the highest occurrence at 35 m. The F. sayana crabs of various sizes could be found solitary or forming aggregations of up to 14 individuals per host, showing no sex or size segregation. In spite of the high diversity of shell species occupied by the hermit crabs and also available in the field, only a few of them were also utilized by P. sayana. The majority (93%) of shells utilized by P. sayana also hosted other symbiont species, constituting the basis of extensive symbiotic complexes. Thus, the ecological relationship between D. insignis and P. sayana may be classified as a non-obligate and non-specific symbiosis that may also involve other facultative organisms such as sea anemones. (C) 2008 Elsevier B.V. All rights reserved.

Validation of a mutant of the pore-forming toxin sticholysin-I for the construction of proteinase-activated immunotoxins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ecologia de actiniários no entremarés rochoso: mensuração do tamanho, estrutura da comunidade e avaliação do deslocamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Counts of benthos communities in Kongsfjorden in 2009

In order to reveal the structure of the sparsely known deeper sublittoral hard bottom communities of glacial Kongsfjorden, the macroepibenthos from six depth zones (30-200 m) was analysed. A total of 180 still images derived from 6-h video recorded at the Kongsfjordneset remotely operated vehicle station were assessed quantitatively. Overall 27 mainly suspension-feeding species/taxa were observed. Of these, two-thirds have an arcto-boreal distribution, while the remainder are cosmopolitan. The overall mean epibenthos abundance was 33 ind./m**2 with maximum values at 150 m depth (97.9 ind./m**2). The majority of the taxa inhabited the entire depth range. Encrusting red algae, an unidentified sponge and the sea anemone Urticina eques, characterized the assemblage of the shallow zone. The sea anemones Hormathia spp. were important below 30 m, the Serpulid polychaete Protula tubularia was characteristic for the community below 50 m and the demosponge Haliclona sp. was a key taxon between 100 and 200 m depth. Cluster analysis and non-metrical multidimensional scaling based on abundance data showed differences between the assemblages along the bathymetric gradient, but only in the shallower depths in relation to the substratum surface incline. As surface and tidal current impacts attenuate with increasing depth, there is a gradual trend from robust key species towards more fragile ones (i.e. P. tubularia), in line with the 'Physical control hypothesis'.