Morphological and functional study of the marginal sphincter of the sea anemones Phymactis clematis and Aulactinia marplatensis from intertidal of Mar del Plata, Argentina

It was made the characterization of marginal sphincter to the species Phymactis clematis (Drayton in Dana, 1849) and Aulactinia marplatensis (Zamponi, 1977), from intertidal ecosystem through their morphogical and functional study. The species P. clematis has a circumscript sphincter of palmate type. This muscle is constituted by a mesogloeal axis and several mesogloeal subaxes. Axis as well as subaxes give a support to the endoderm which border is smooth. Aulactinia marplatensis has a circunscript sphincter pinnate type. The axis has a truncated cone shape while in P. clematis the shape is cylindrical on its origin and it is bifurcated at the end. Both species experiments were carried out using the isolated muscles. They were stimulated at increasing KCl concentrations ranging from 20 to 200 mM. The results were analysed in the form of dose-response curves expressed in tension in grams force vs concentration. Contractil force increases in a sigmoid form to increasing KCl concentrations. The correlation between morphology and function and the differences shown in both species would be related to their intertidal distribution.

Partial sequence and toxic effects of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera

A neurotoxic peptide, granulitoxin (GRX), was isolated from the sea anemone Bunodosoma granulifera. The N-terminal amino acid sequence of GRX is AKTGILDSDGPTVAGNSLSGT and its molecular mass is 4958 Da by electrospray mass spectrometry. This sequence presents a partial degree of homology with other toxins from sea anemones such as Bunodosoma caissarum, Anthopleura fuscoviridis and Anemonia sulcata. However, important differences were found: the first six amino acids of the sequence are different, Arg-14 was replaced by Ala and no cysteine residues were present in the partial sequence, while two cysteine residues were present in the first 21 amino acids of other toxins described above. Purified GRX injected ip (800 µg/kg) into mice produced severe neurotoxic effects such as circular movements, aggressive behavior, dyspnea, tonic-clonic convulsion and death. The 2-h LD50 of GRX was 400 ± 83 µg/kg.

Behavioral and electroencephalographic analysis of seizures induced by intrahippocampal injection of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera

In this study, the behavioral and electroencephalographic (EEG) analysis of seizures induced by the intrahippocampal injection in rats of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera, was determined. The first alterations occurred during microinjection of granulitoxin (8 µg) into the dorsal hippocampus and consisted of seizure activity that began in the hippocampus and spread rapidly to the occipital cortex. This activity lasted 20-30 s, and during this period the rats presented immobility. During the first 40-50 min after its administration, three to four other similar short EEG seizure periods occurred and the rats presented the following behavioral alterations: akinesia, facial automatisms, head tremor, salivation, rearing, jumping, barrel-rolling, wet dog shakes and forelimb clonic movements. Within 40-50 min, the status epilepticus was established and lasted 8-12 h. These results are similar to those observed in the acute phase of the pilocarpine model of temporal lobe epilepsy and suggest that granulitoxin may be a useful tool not only to study the sodium channels, but also to develop a new experimental model of status epilepticus.

Identification of hemolytic and neuroactive fractions in the venom of the sea anemone Bunodosoma cangicum

Sea anemones are a rich source of biologically active substances. In crayfish muscle fibers, Bunodosoma cangicum whole venom selectively blocks the I K(Ca) currents. In the present study, we report for the first time powerful hemolytic and neuroactive effects present in two different fractions obtained by gel-filtration chromatography from whole venom of B. cangicum. A cytolytic fraction (Bcg-2) with components of molecular mass ranging from 8 to 18 kDa elicited hemolysis of mouse erythrocytes with an EC50 = 14 µg/ml and a maximum dose of 22 µg/ml. The effects of the neuroactive fraction, Bcg-3 (2 to 5 kDa), were studied on isolated crab nerves. This fraction prolonged the compound action potentials by increasing their duration and rise time in a dose-dependent manner. This effect was evident after the washout of the preparation, suggesting the existence of a reversible substance that was initially masking the effects of an irreversible one. In order to elucidate the target of Bcg-3 action, the fraction was applied to a tetraethylammonium-pretreated preparation. An additional increase in action potential duration was observed, suggesting a blockade of a different population of K+ channels or of tetraethylammonium-insensitive channels. Also, tetrodotoxin could not block the action potentials in a Bcg-3-pretreated preparation, suggesting a possible interaction of Bcg-3 with Na+ channels. The present data suggest that B. cangicum venom contains at least two bioactive fractions whose activity on cell membranes seems to differ from the I K(Ca) blockade described previously.

Distribuição geográfica da fauna e flora da Baía de Guanabara

The author studied, the horizontal and vertical distribution of most common part of the flora and fauna of the bay of Guanabara at Rio de Janeiro. In this paper the eulittoral, poly, meso and oligohaline regions were localised and studied; and the first chart of its distribution was presented (fig. 2). The salinity of superficial waters was established through determinations based on 30 trips inside the buy for collecting biological materials. Some often 409 determinations which were previous reported together with the present ones served for the eleboration of a salinity map of the bay of Guanabara (fig. 1). This map of fig. 2 shows the geographic locations of the water regions. EULITTORAL WATER REGIME — Fig. 3 shows the diagram scheme of fauna and flora of this regime. Sea water salinity 34/1.000, density mean 1.027, transparent greenish waters, sea coast with moderate bursting waves. Limpid sea shore with white sand, gneiss with the big barnacle Tetraclita squamosa var. stalactifera (Lam. Pilsbry. Vertical distributions: barna¬cles layers with a green region in which are present the oyster Ostrea pa-rasitica L., the barnacles Tetraclita, Chthamalus, Balanus tintinnabulum var. tintinnabulum (L.) e var. antillensis Pilsbry in connection with several mollusca and the sea beatle Isopoda Lygia sp. Covered by water and exposed to air by the tidal ritms, there is a stratum of brown animals that is the layer of mussels Mytilus perna L., with others brown and chestnut animals : the Crustacea Pachygrapsus, the little crab Porcellana sp., the stone crab Me-nippe nodifrons Stimpson, the sea stars Echinaster brasiliensis (Mull. & Tr.), Astropecten sp. and the sea anemones Actinia sp. Underneath and never visible there is a subtidal region with green tubular algae of genus Codium and amidst its bunches the sea urchin Lycthchinus variegatus (Agass.) walks and more deeply there are numerous sand-dollars Encope emarginata (Leske). The microplancton of this regime is Ceratiumplancton. POLYHALINE WATER REGIMB — Water almost sea water, but directly influenced by continental lands, with rock salts dissolved and in suspension. Salinity: 33 to 32/1.000. This waters endure the actions of the popular nicknamed «water of the hill» (as the waters of mesohaline and oligohaline regimes), becoming suddenly reddish during several hours. That pheno¬menon returns several times in the year and come with great mortality of fishes. In these waters, according to Dr. J. G. FARIA there are species of Protozoa : Peridinea, the Glenoidinium trochoideum St., followed by its satellites which he thinks that they are able to secret toxical substances which can slaughter some species of fishes. In these «waters of the hill» was found a species of Copepoda the Charlesia darwini. In August 1946 the west shore of the Guanabara was plenty of killed fishes occupying a area of 8 feet large by 3 nautical miles of lenght. The enclosure for catching fishes in the rivers mouthes presents in these periods mass dead fishes. The phenomenon of «waters of the hill» appears with the first rains after a period of long dryness. MESOHALINE WATER REGIME — Fig. 4 shows the the diagramm scheme. Salt or brackish water from 30 to 17/1.000 salinity, sometimes until 10/1.000. Turbid waters with mud in suspension, chestnut, claveyous waters; shore dirty black mud without waving bursting; the waters are warmer and shorner than those of the polihaline regime. Mangrove shore with the mangrove trees : Rhizophora mangle L., Avicennia sp., Laguncularia sp., and the »cotton tree of sea» Hibiscus sp. Fauna: the great land crab «guaimú» Cardisoma guanhumi Latr., ashore in dry firm land. There is the real land crab Ucides cordatus (L.) in wetting mud and in neigh¬ bourhood of the burrows of the fiddler-crabs of genus Uca. On stones and in the roots of the Rhizophora inhabits the brightly colored mangrove-tree-crab («aratu» Portuguese nickname) Goniopsis cruentata (Latreille) and the sparingly the big oyster Ostrea rhizophorae Guild. Lower is the region of barnacles Balanus amphitrite var. communis Darwin and var. niveus Darwin; Balanus tintinnabulum var. tintinnabulum (L.) doesn't grow in this brackish water; lower is the region of Pelecipoda with prepollency of Venus and Cytherea shell-fishes and the Panopeus mud crab; there are the sea lettuce Ulva and the Gastreropod Cerithium. The Paguridae Clibanarius which lives in the empty shells of Gasteropod molluscs, and the sessile ascidians Tethium plicatum (Lesuer) appears in some seasons. In the bottom there is a black argillous mud where the «one landed shrimps» Alpheus sp. is hidden. OLIGOHALINE WATER REGIME — The salinity is lower than 10/1.000. average 8/1.000. There are no barnacles and no sea-beetles Isopods of genus Lygia; on the hay of the shore there are several graminea. This brackish water pervades by mouthes of rivers and penetrates until about 3 kilometers river above. While there is some salt dissolved in water, there are some mud crabs of the genus Uca, Sesarma, Metasesarma and Chasmagnatus. The presence of floating green plants coming from the rivers in the waters of a region indicated the oligohaline waters, with low salt content because when the average of NaCl increases above 8/1.000 these plants die and become rusty colored.

Sea urchin granuloma

Injuries caused by venomous and poisonous aquatic animals may provoke important morbidity in humans. The phylum Echinoderma include more than 6000 species of starfish, sea urchins, sand dollars, and sea cucumbers some of which have been found responsible for injuries to humans. Initial injuries by sea urchins are associated with trauma and envenomation, but later effects can be observed. Sea urchin granuloma is a chronic granulomatous skin disease caused by frequent and successive penetration of sea urchin spines which have not been removed from wounds. The authors report a typical case of sea urchin granuloma in a fisherman and its therapeutic implications.

SURVIVAL, INDUCTION AND RESUSCITATION OF Vibrio cholerae FROM THE VIABLE BUT NON-CULTURABLE STATE IN THE SOUTHERN CARIBBEAN SEA

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Observation of initial clinical manifestations and repercussions from the treatment of 314 human injuries caused by black sea urchins (Echinometra lucunter) on the southeastern Brazilian coast

INTRODUCTION: Injuries caused by sea urchins are the most common caused by marine animals in humans in Brazil, with the black sea urchin (Echinometra lucunter) causing the most injuries to bathers. METHODS: This study observed 314 human wounds with emphasis on the early observation of clinical signs and symptoms and their implications on the recommended treatment. RESULTS: All the injuries were caused by black sea urchins and were observed in bathers. The lesions and the pain were associated with penetration of the spines; there was no early inflammation or pain without pressure on the wounded places. Complications arising from this kind on injury, including infections and foreign body granulomas, are associated with the permanence of the spines in the wounds. CONCLUSIONS: The study confirmed that this kind of injury is the most common accident caused by aquatic animals in Brazil. The main therapeutical recommendation is early removal of the spines to prevent late complications, such as infections and the formation of foreign body granulomas.

The diet of the South American sea lion (Otaria flavescens) at Río Negro, Patagonia, Argentina, during the winter-spring period

The South American sea lion, Otaria flavescens (Shaw, 1800) population is steadily expanding along the Patagonian coast of Argentina in the last decades. However, little is known about the feeding ecology of the species in the area. The aim of this study was to analyze the food habits of O. flavescens from 91 scats collected at Río Negro province, during the winter and spring of 2005. Fish occurred in 96% of scats containing prey remains, followed by cephalopods (26%). Raneya brasiliensis (Kaup, 1856) was the most frequent and abundant species occurring in 58.6% of samples and constituting almost 50% of fish predated. Second in importance were Porichthys porosissimus (Cuvier, 1829) and Cynoscion guatucupa (Cuvier, 1830) in terms of occurrence (%FO 20.7) and numbers (29.6%) respectively. The squid Loligo gahi (d'Orbigny, 1835) was the most frequent cephalopod prey (42.1%), whereas Octopus tehuelchus (d'Orbigny, 1834) was the most abundant (77%). The higher amount and diversity of prey found in the spring in comparison with the winter season might be related to a higher feeding activity of seals or to a seasonal increase in food availability in the area.

Plastic ingestion by sea turtles in Paraíba State, Northeast Brazil

ABSTRACT Currently, plastics are recognized as a major pollutant of the marine environment, representing a serious threat to ocean wildlife. Here, we examined the occurrence and effects of plastic ingestion by sea turtles found stranded along the coast of Paraíba State, Brazil from August 2009 to July 2010. Ninety-eight digestive tracts were examined, with plastic found in 20 (20.4%). Sixty five percent (n = 13) of turtles with plastic in the digestive tract were green turtles (Chelonia mydas), 25% (n = 5) were hawksbills (Eretmochelys imbricata), and 10% (n = 2) were olive ridley (Lepidochelys olivacea). More plastic was found in the intestine (85%) than in other parts of the gastrointestinal tract. We observed complete blockage of the gastrointestinal tract due to the presence of plastic in 13 of the 20 turtles that had ingested plastic. No correlation was found between the curved carapace length (CCL) and the number or mass of the plastic ingested items. Significant differences were found between the intake of hard and soft plastic and the ingestion of white/transparent and colored plastic, with soft and white/transparent plastics being more commonly ingested. This study reveals the serious problem of plastic pollution to sea turtles at the area.

Parasites of commercial shrimps and fishes in Argentine sea: on the adult and metacercaria of Opecoeloides feliciae n. sp. (Digenea: Opecoelidae)

Opecoeloides feliciae n. sp., first record of opecoelid metacercariae in commercial shrimps of South Atlantic Ocean, parasitizing Cynoscion striatus (adults) and Artemesia longinaris (metacercariae), is described, illustrated and compared with related species of the genus. Adults, immature worms and metacercariae are compared, and rates of prevalence and intensity of infection are also given.

Morphological and molecular characterisation of Heliconema hainanensis sp. nov. (Spirurina: Physalopteridae) from congers in the South China Sea, with a key to the species of Heliconema

Heliconema hainanensis sp. nov. collected from Uroconger lepturus (Richardson) (Anguilliformes: Congridae), Muraenesox cinereus (Forsskål) and Congresox talabonoides (Bleeker) (Anguilliformes: Muraenesocidae) in the South China Sea was described using light and scanning electron microscopy. The new species differs from its congeners by the following morphology: pseudolabia, the number and arrangement of caudal papillae (4 pairs of pedunculate precloacal papillae arranged in 2 groups of 2 and 2 pairs and 6 pairs of pedunculate postcloacal papillae arranged in 4 groups of 1, 2, 1 and 2 pairs), the length of spicules [left spicule 0.51-0.69 mm, right spicule 0.20-0.27 mm, spicule (right:left) ratio 1:2.20-2.69] and the morphology of the female tail tip. In addition, specimens of the new species collected from the three different hosts and specimens of an unidentified species of Heliconema collected from U. lepturus were characterised using molecular methods by sequencing the internal transcribed spacer (ITS) of ribosomal DNA. Analyses and comparison of the ITS sequence of H. hainanensis sp. nov. with Heliconema sp. support the validity of the new species based on morphological observations. An identification key to the species of Heliconema is also provided.

Modification of pyruvate kinase and lactate dehydrogenase in foot muscle of the sea mussel Mytilus galloprovincialis under anaerobiosis and recovery

The modification of pyruvate kinase (PK) and lactate dehydrogenase (LDH) activity in foot muscle of the mussel Mytilus galloprovincialis during exposure to air and recovery in water was investigated. In the course of exposure to air, the activity of these enzymes measured at high and low substrate concentrations showed successive increases and decreases. Returning the mussels to water after exposure to air affected enzyme activity in a manner similar to anaerobiosis. When measuring at saturated concentrations of substrates and substrate and coenzyme for PK and LDH, respectively, the maximum activation of PK (37%) was observed at 4 h of animal exposure to air, and for LDH (67%) at 6 h exposure to air. During 24 h of exposure of animals to air, PK activity practically reached the stock level, while LDH was still activated (148%). The change in lactate dehydrogenase activity in mussel muscle during anoxia and recovery is described here for the first time. Variation in pyruvate kinase activity during exposure to air and recovery is linked to the alteration of half-maximal saturation constants and maximal velocity for both substrates. The possible role of reversible phosphorylation in the regulation of pyruvate kinase and lactate dehydrogenase properties is discussed

Toxic, antimicrobial and hemagglutinating activities of the purple fluid of the sea hare Aplysia dactylomela Rang, 1828

The antimicrobial, hemagglutinating and toxic activities of the purple fluid of the sea hare Aplysia dactylomela are described. Intact or dialyzed purple fluid inhibited the growth of species of Gram-positive and Gram-negative bacteria and the action was not bactericidal but bacteriostatic. The active factor or factors were heat labile and sensitive to extreme pH values. The fluid preferentially agglutinated rabbit erythrocytes and, to a lesser extent, human blood cells, and this activity was inhibited by the glycoprotein fetuin, a fact suggesting the presence of a lectin. The fluid was also toxic to brine shrimp nauplii (LD50 141.25 µg protein/ml) and to mice injected intraperitoneally (LD50 201.8 ± 8.6 mg protein/kg), in a dose-dependent fashion. These toxic activities were abolished when the fluid was heated. Taken together, the data suggest that the activities of the purple fluid are due primarily to substance(s) of a protein nature which may be involved in the chemical defense mechanism of this sea hare.