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A closed marine culture system for rearing Octopus joubini and other large-egged benthic octopods.

The system consists of 2 adjoining 150 litre aquaria, one functioning as the water-conditioning tank and the other as the principal rearing tank. Water quality remained high with biological and mechanical filtration, physical adsorption and ultraviolet-light disinfection taking place exclusively in the conditioning tank. The pH ranged from 7.58 to 8.00 and ammonia and nitrite levels never exceeded 0.004 mg/l and 0.198 mg/l, respectively. Nitrate levels were maintained at 40 mg/l or less with no adverse affects. Adult octopuses readily mated and females produced 50-150 eggs, with 95% hatching success. When fed small liver crabs, the octopus hatchlings were reared to sexual maturity either in groups or individually in about 120-150 days. Growth rates (4% bodyweight/day) and food conversion efficiences (30-40%) were as high as those obtained in open systems by previous workers.

Ammonia↗

Methods for the aquarium maintenance of the common octopus of British waters, Eledone cirrhosa.

Healthy, undamaged specimens survive well in recirculating aquarium seawater of about 36 parts per thousand salinity and PH 7.4, having 50 mg.1(-1) nitrogen as nitrate, less than 0.1 mg.1(-1) nitrogen as nitrite, nitrogen as nitrate, and less than 0.1 mg.1(-1) nitrogen as ammonia, and a m ean annual temperature of 14-15 degree C, about 5 degree C above ambient. For maximal growth rates, the gross wet weight of liver crabs required as food ranges up to 10% of the weight of the octopus. Weight-specific growth rates fall from 3.4% day-1 at 100-200 g bodyweight, to 1-1.5% day-1 at greater than 500 g bodyweight. Survival of healthy, wild-caught animals, commonly 4.6 months and up to 8 months, is apparently limited more by endogenous factors concerned with sexual maturation and lifespan than by aquarium conditions. Eggs have been laid but it has not yet been possible to hatch and rear them.

Anesthesia↗

Effects of dietary shrimp, squid and octopus on serum and liver lipid levels in mice.

The effects of three seafoods, shrimp, squid and octopus, on lipid metabolism were investigated in mice fed on 0.1% and 1.0% cholesterol-supplemented diets in the first experiment. One of each of these seafoods and casein were added to the basal diet at levels of 15% and 5%, respectively, as proteins. Casein served as the sole protein source of the control diet. The serum cholesterol concentration was significantly lower in the mice fed on shrimp and squid in the 0.1% cholesterol diet and on any seafood in the 1.0% cholesterol diet when compared with that in the mice fed on the control diet. The liver cholesterol concentration was significantly lower in all seafood groups given the 0.1% cholesterol diet, and in the squid and octopus groups given the 1.0% cholesterol diet. In the second experiment, the effect of these seafoods on lipid metabolism was compared with that of their defatted products in mice fed on a 0.2% cholesterol diet. Defatting resulted in an increase in the serum cholesterol and triglyceride levels in the shrimp and squid groups. The hepatic cholesterol concentration in all the seafood groups was significantly lower than that in the control group, and defatting did not influence the liver cholesterol concentration. Fecal total steroid excretion was higher in all the seafood groups when compared with that in the control group, and was not modified by the removal of fats. Thus, shrimp, squid and octopus exerted hypolipidemic activity; the serum cholesterol-lowering activity of shrimp and squid was attributed to their lipid fraction, whereas the non-lipid fraction of shrimp, squid and octopus contributed to a reduction of hepatic cholesterol and an increase of fecal steroid excretion.

Animal Nutritional Physiological Phenomena↗

Quantification and speciation of mercury and selenium in fish samples of high consumption in Spain and Portugal.

Mercury (Hg) and selenium (Se) determinations were carried out to evaluate human exposure to those elements through fish consumption in Spain and Portugal. Atomic fluorescence spectroscopy (AFS) was applied in a cold vapor mode for total mercury quantification and was also hyphenated to gas chromatography (GC) to achieve the speciation of organomercurial species in fish samples. The results obtained show the highest concentration of Hg in swordfish and tuna (0.47+/-0.02 and 0.31+/-0.01 microg g-1, respectively), and a much lower concentration in sardine, mackerel shad, and octopus (0.048+/-0.002, 0.033+/-0.001, and 0.024+/-0.001 microg g-1, respectively). The determination of alkyl mercury compounds revealed that 93-98% of mercury in the fish samples was in the organic form. Methylmercury (MeHg) was the only species found in the three fish species with higher mercury content. Total selenium concentration was high in sardine, swordfish, and tuna (0.43+/-0.02, 0.47+/-0.02, and 0.92+/-0.01 microg g-1, respectively), but low in mackerel shad and octopus (0.26+/-0.01 and 0.13+/-0.01 microg g-1, respectively). Speciation of selenium compounds was done by high-performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (LC-ICP-MS). Selenomethionine (SeMet) was the only selenium compound identified in the fish samples with higher selenium content. Among the fish species studied, sardine had the most favourable Se:Hg and SeMet:MeHg molar ratios; therefore, its consumption seems to be preferable.

Animals↗

Phosphoglycopeptide, a major constituent of the spermatophoric plasma of the octopus (Octopus dofleini martini).

A phosphoglycopeptide, accounting for approximately 90% of the characteristically high content of acid-soluble organically-bound phosphorus in the octopus spermatophoric plasma (4 mg P/ml), was identified. Electrophoretic and chromatographic purification, followed by chemical and enzymic hydrolysis, yielded D-galactose phosphate as a degradation product. The galactose and peptide moieties of the compound were linked via a phosphoryl rather than a glycosidic linkage but the peptide was devoid of aromatic amino acids.

Amino Acids↗

Two new Aggregata species (Apicomplexa: Aggregatidae) infecting Octopus tehuelchus and Enteroctopus megalocyathus (Mollusca: Octopodidae) in Patagonia, Argentina.

During a long-term study carried out between 1981 and 1996 on the biological and fishery aspects of octopuses inhabiting the Gulfs of San Matías, San José, and Nuevo, Patagonia, Argentina, 2 new species of Aggregata (Apicomplexa: Aggregatidae) were found in the digestive tracts of Octopus tehuelchus d'Orbigny, 1834 (prevalence 72%) and Enteroctopus megalocyathus (Gould, 1852) (prevalence 77%). Both species can be distinguished from other congeners on the basis of their hosts, the diameters of sporocysts, and number and length of sporozoites. Despite overlap in the distributions of the two hosts species in the area covered in this study, both Aggregata new species exhibited high host specificity.

Animals↗

Dicyemennea canadensis n. sp. (phylum Dicyemida) from Bathypolypus arcticus (Mollusca: Cephalopoda: Octopoda).

Dicyemennea canadensis n. sp. is described from a bathyal octopus collected off Canada in the Bay of Fundy. The dicyemid is a small species that rarely exceeds 600 microm in length. The vermiform stages are further characterized as having 17-23 peripheral cells, a conical-shaped calotte, an axial cell that extends to the base of the propoplar cells, and no abortive axial cells. Infusoriform embryos consist of 37 cells. There is 1 nucleus in each urn cell, and refringent bodies are absent. This is the first dicyemid to be described from the cephalopod Bathypolypus arcticus (Prosch. 1847), and the first dicyemid reported from Canada. In addition, it is the first species of Dicyemennea from the northwestern Atlantic Ocean to be described.

Animals↗

Dicyema shorti n. sp. (phylum Dicyemida) from Octopus burryi (Mollusca: Cephalopoda: Octopodidae) in the Gulf of Mexico.

Nematogens and vermiform embryos of a new species of Dicyema are described from an octopus collected off Veracruz, Mexico. Dicyema shorti n. sp. is a small dicyemid species that rarely exceeds 500 microm in length. It is further characterized by the presence of 18 peripheral cells in the vermiform stages, a conical-shape calotte, and an axial cell that extends to the base of the propolar cells. Other stages in the life cycle of the parasite are not known. This is the first dicyemid to be described from Octopus burryi Voss. 1950, and also from both the southern Gulf of Mexico and the country of Mexico.

Animals↗

New species of Dicyemennea (phylum: Dicyemida) in deep-water Graneledone (Mollusca: Cephalopoda: Octopoda) from the Antarctic.

Two new species of dicyemids are described from 2 species of deep benthic cephalopods, Graneledone antarctica and G. macrotyla, collected in the Southern Ocean south of the Antarctic Convergence. Dicyemennea bathybenthum n. sp. was found in G. antarctica. It is a medium-sized dicyemid whose length does not exceed 1,000 microm. The calotte is bluntly rounded or conical in small individuals, but the shape becomes discoidal in large individuals. Nematogens and vermiform embryos have 23 peripheral cells. An anterior abortive axial cell is present in vermiform embryos. Other stages in the life cycle were not observed. Dicyemennea dorycephalum n. sp. was found in G. macrotyla. It is a medium to large dicyemid that rarely exceeds 4,000 microm. The calotte is distinctly pointed, similar in shape to a spearhead. Vermiform stages typically have either 25 or 27 peripheral cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos have 37 cells, the refringent bodies are solid in composition, and 2 nuclei are present in each urn cell.

Animals↗

Cloning of Octopus cephalotocin receptor, a member of the oxytocin/vasopressin superfamily.

We reported that the common octopus, Octopus vulgaris, in common with vertebrates, possesses two members of the oxytocin/vasopressin superfamily: octopressin (OP) and cephalotocin (CT). This was the first observation of its kind in invertebrates. As OP and CT have different biological activities, the presence of specific receptors has been proposed. We cloned the cDNA of an orphan receptor from Octopus brain and found it to encode a polypeptide of 397 amino acids that displays sequences characteristic of G-protein coupled receptors. The orphan receptor showed high homology to receptors of the oxytocin/vasopressin superfamily and seemed to conserve the agonist-binding pocket common to the oxytocin and vasopressin receptors. Xenopus oocytes that express the orphan receptor responded to the application of CT by an induction of membrane Cl(-) currents coupled to the inositol phosphate/Ca(2+) pathway. OP and the other members of the oxytocin/vasopressin superfamily did not activate this receptor. HPLC fractionation of the Octopus brain extract combined with an oocyte assay yielded a single substance that was identical to CT. On the basis of these results, we conclude that the cloned receptor is the CT receptor (CTR). Expression of CTR mRNA in Octopus was detected in the central and the peripheral nervous systems, the pancreas, the oviduct and the ovary. This receptor may mediate physiological functions of CT in Octopus such as neurotransmission, reproduction and metabolism.

Amino Acid Sequence↗

Cloning, expression and pharmacological characterization of a vasopressin-related receptor in an annelid, the leech Theromyzon tessulatum.

In annelids, it has been established that arginine-vasopressin (AVP)/oxytocin (OT) superfamily peptides are involved in the maintenance of water and electrolyte homeostasis as well as reproduction. At present, there is little information on their receptors. In this study, we report the characterization of a 1.7 kb cDNA for an AVP-related receptor from the leech Theromyzon tessulatum. The open reading frame encodes a 435-aminoacid transmembrane protein that displays seven segments of hydrophobic amino acids, typical of G-protein-coupled receptors. The overall predicted protein exhibits about 30% amino-acid identities to other invertebrate, as well as vertebrate, AVP/OT receptor family members, and displays conserved characteristic features belonging to the AVP/OT receptor superfamily. RT-PCR expression experiments showed that mRNA is expressed in the genital tract, the ovary and the brain. The receptor expression is stage specific, showing a weak expression after the two first blood meals, increasing dramatically after the last blood meal during the period of sexual maturation and disappearing after egg laying. Thus, the leech AVP-related receptor may mediate reproductive functions. When expressed in COS-7 cells, the receptor binds ligands with the following rank order of potency: AVP= Arg-vasotocin >Arg-conopressin >mesotocin = OT = Lys-conopressin=isotocin>annetocin. This shows an AVP-like pharmacological profile. The transfected receptor mediates AVP-induced accumulation of inositol phosphates, indicating that the leech AVP-related receptor is functional. This study describes the characterization of a novel AVP/OT superfamily receptor in annelids, which are considered the most distant group of coelomate metazoans possessing a functional AVP/OT-related endocrine system.

Amino Acid Sequence↗

Anti-bovine rhodopsin monoclonal antibody recognizing light-dependent structural change.

The antigenic structure of the bovine rhodopsin molecule was investigated by using a bovine rhodopsin-specific monoclonal antibody designated Rh 29. Competition assay with sealed intact disks and broken disks indicated that the antibody-binding region was localized in the intradiscal surface. An antigenic peptide obtained by a cyanogene bromide cleavage of rhodopsin was purified and determined as residues 2-39 in the amino acid sequence. Further analysis suggested that the antigenic determinant included at least residues 21-25. These results were consistent with the structural model for membrane topology of rhodopsin. The antigenicity of the rhodopsin was compared among several states. The antibody bound to both ammonyx LO-solubilized unbleached and bleached rhodopsin. In contrast, upon membrane-embedded rhodopsin, unbleached one was 100-times less antigenic than bleached one. The results suggested that the segment around the determinant of membrane-embedded rhodopsin should undergo a structural change upon absorption of light. Rh 29 detected a band corresponding to bovine, porcine and octopus opsins in immunoblotting. Protein blot of crayfish rhabdome did not show any reactive band. These bands except for crayfish reacted with concanavalin A as well. The N-terminal structure may, therefore, conserved between mammal and erthropoda and diverge between them and cepharopoda.

Amino Acid Sequence↗

Post-hatching development of the brain in Octopus ocellatus.

To investigate the post-hatching development of the brain in a benthic octopod, Octopus ocellatus, we performed volumetric analyses of the brain. The brain consisting of the supra- and subesophageal masses was divided into 5 regions according to the functions suggested for the brain of another benthic octopod Octopus vulgaris, and the volume of each region was estimated at three post-hatching ages. We found that the inferior frontal lobe system and the brachial lobe increased in relative volume as the animals grew, while the basal lobe system decreased in relative volume. This result suggests that increasing demand for processing tactile information after hatching is reflected in the higher developmental rate in the centers devoted for tactile sense and related learning. We also found that the inner neuropile layer mainly consisting of dendrites, synapses and axons showed great increases in volume compared with the outer neural-cell-body layer. Although the increase in volume of the inner layer was marked during 1 month after hatching in all brain regions examined, the extent of the increase varied among brain regions. Developmental changes in cell densities in the outer layer also varied among the regions. The present results suggest that the post-hatching development of the brain in O. ocellatus is not homogeneous but varies among brain regions depending on different roles in controlling the behavior.

Animals↗