Algal phospholipids by 31P NMR: Comparing isopropanol pretreatment with simple chloroform/methanol extraction

• 1.1. A modified Folch procedure [potassium (ethylenedinitrilo)-tetraacetic acid at pH 6 substituted for KC1] is suitable for the extraction of marine algae.
• 2.2. The quantitative ³¹P nuclear magnetic resonance phospholipid profiles of four marine algae, Gracilaria verrucosa, Bryothamnion triquetrum, Padina gymnospora, and Caulerpa sertularioides, were obtained from Folch and Nichols extractions of both fresh and dried algae, and essentially identical results were obtained using either extraction procedure.
• 3.3. Extracts of air-dried algae are statistically different when compared to extracts of living algae, suggesting that tissue handling is a critical factor in phospholipid extractions.

     

Specific dynamic action demonstrated in the herbivorous marine periwinkles,LIttorina littorea L. and Littorina obtusata L. (Mollusca,Gastropoda)

• 1.1. Respiration rates of starved periwinkles, Liltorina littorea and Littorina obtusata, increased by 40–60% when fed their preferred algal food for 1 hr, or after having been exposed for the same period to an aqueous extract of the alga.
• 2.2. The stimulus causing the rise in metabolic rate by algal extracts is therefore thought to be chemosensory in nature, and possibly composed primarily of dissolved free amino acids.
• 3.3. The respiration rates of L. littorea responded only to the green alga Enteromorpha intestinalis while L. obtusata demonstrated an increase in respiration rates when fed the ubiquitous brown alga, Ascophyllum nodosum, and to a lesser degree to Enteromorpha.
• 4.4. These results compare very well to postprandial increases in oxygen consumption demonstrated in vertebrates and marine bivalves where the components of specific dynamic action (SDA) are thought to be chiefly biosynthetic costs of digestion and assimilation.

     

A comparative study of seafish erythrocytes and agglutinins from seaweeds

• 1.1. To date, only a few authors have assayed the agglutinic activity of marine algae against fish erythrocytes, and in these cases, mainly against freshwater fish.
• 2.2. For the first time, the hemagglutinic activity of 70 seaweeds (29 brown, 37 red and four green algae) against erythrocytes of 16 seaflsh species is reported.
• 3.3. The presence of agglutinins was demonstrated in 100% of algae assayed, against at least one of the different types of erythrocytes tested.
• 4.4. The results obtained confirm the presence of receptors for algae agglutinins on the surface of the erythrocytes of the fish studied.
• 5.5. This could be useful in establishing the origins of fish populations, as these serological differences could distinguish between populations of cultivated and wild fish.

     

The occurrence of brassicasterol and epibrassicasterol in the chromophycota

• 1.1. Sterols were identified from eight isolates of five species in the Chromophycota that were cultured axenically and harvested in the stationary phase.
• 2.2. Analyses were performed on four strains from the Prymnesiophyceae, two strains from the Cryptophyceae and one from the Bacillariophyceae. Most strains examined contained only one major sterol, 24-methyl-22-dehydrocholesterol.
• 3.3. Analysis by capillary GC, HPLC, and in one instance NMR, showed that the two strains provisionally identified as Isochrysis contained brassicasterol (24β-methyl-22-dehydrocholesterol); whereas, all other species examined contained primarily epibrassicasterol (24α-methyl-22-dehydrocholesterol).
• 4.4. Stigmasterol (24α-ethyl-22-dehydrocholesterol) accompanied epibrassicasterol in Pleurochrysis carterae.
• 5.5. Analyses of C-24 alkyl isomers in these algae may provide useful information concerning their taxonomic placement.
• 6.6. The occurrence of both isomers of 24-methyl-22-dehydrocholesterol in oysters is explained by the occurrence of both isomers among algae which are probably dietary sources for oysters.

     

Solving parasite-related problems in cultured marine fish

• 1.1. Our knowledge of parasite born e diseases of cultured marine fish is restricted by the limited extent of practiced mariculture.
• 2.2. Several parasites, pathogenic to fish including capsaloid, microcotylid and diclidophoran monogeneans and caligid copepods are ubiquitous in mariculture systems in diverse species of fish and geographical regions.
• 3.3. The alga Amyloodinium ocellatum and the ciliated protozoan Cryptocaryon irritons are not as ubiquitous, but nevertheless present the most serious risk to farmed fish of most species due to their high pathogenicity, fecundity and resistance to conventional means of control.
• 4.4. Infections in maricultured fish are dominated by monoxenous parasites. Occurence of heteroxenous parasites is restricted by their requirement for additional hosts to maintain transmission.
• 5.5. Cage culture allows free exchange of infection between caged and native fishes; in inshore systems infections have to be introduced primarily from external sources.
• 6.6. Water supply does not appear to be an efficient route of entry of infection into culture systems; parasites are introduced either by wild fry or wild breeders collected into the system, or with wild fish infiltrating via water supply or drainage.
• 7.7. Adverse growth conditions are the most common circumstances in outbreak of epizooites. Over-crowding does not appear to promote primarily epizooites, its impact is indirect by contributing to the deterioration of the growth conditions.
• 8.8. Epizooites are likely to occur also in juvenile, immunologically naive fish as well as following introduction of parasites into unexperienced fish population.
• 9.9. Control has been implemented thus far through an empirical approach. Recent research is geared towards the development of methodology specifically designed to the needs of mariculture.

     

Selective activation of carcinogenic aromatic amines to bacterial mutagens in the marine musselMytilus gallopro vincialis

• 1.1. The postmitochondrial fraction of the marine mussel Mytilus galloprovincialis digestive gland activates selectively precarcinogenic aromatic amines, but not precarcinogenic benzo(a]pyrene, to Salmonella typhimurium TA 98 mutagens.
• 2.2. This activation potential is NADPH-dependent, is not inducible by exposure to Diesel 2 oil and a polluted environment, and is inhibited by methimazole.
• 3.3. The characteristics of this activation potential are consistent with the recent finding of the presence of FAD-containing-, and lack ofcytochrome P-450 dependent-, monooxygenase activity in Mytilus edulis.
• 4.4. The presence of such selective potential in marine invertebrate(s) may bring new insight into our understanding of the fate and the effects of carcinogens in the marine environment.

     

Organohalogen constituents of the acorn worm Ptychodera bahamensis

• 1.1. Twenty organohalogen compounds, primarily phenols, were detected in the volatile extracts of the acorn worm Ptychodera bahamensis.
• 2.2. Five chlorinated compounds, previously undescribed from acorn worms, were identified.
• 3.3. Enteropneusts can be significant contributors of halogenated organics to the marine environment.

     

Ferredoxin-glutamate synthase from Chlamydomonas reinhardii. Prosthetic groups and preliminary studies of mechanism

• 1.1. Ferredoxin-glutamate synthase from the green alga Chlamydomonas reinhardii appears to contain as prosthetic groups, 1 FAD, 1 FMN and 1 |3Fe-xS | cluster, per molecule of M_r = 146,000.
• 2.2. The synthesis of glutamate, catalyzed by this enzyme, proceeds through the formation of an enzyme-bound free radical of flavin semiquinone.
• 3.3. This enzyme may catalyze the ferredoxin-dependent reduction of 2-oxoglutarate, in the absence of glutamine.

     

Sterol composition of three marine sponge species from the genus Cinachyrella

• 1.1. The hitherto undescribed sterol compositions of three marine sponge species belonging to the genus Cinachyrella are reported: C. alloclada and C. kükenthali from the Senegalese coast, at two different depths, and C. aff. schulzei from the lagoon of Nouméa, New Caledonia.
• 2.2. Fourteen free sterols have been identified by GC and GC/MS studies, including the 23,24ξ-dimethylcholesta-5,22-dien-3β-ol (10) and the rare 24-norcholesta-5,22-dien-3β-ol (1).
• 3.3. The first compound (10) is reported for the second time in a marine sponge and it was found only in Senegalese sponges collected in shallow waters.
• 4.4. Sterol (10) has been isolated by HPLC and identified by NMR techniques.
• 5.5. Significant amounts of cholest-7-en-3β-ol (7) were also found in the Senegalese sponge species.
• 6.6. Apart from these two compounds, the three sponge sterol compositions are found to be very similar.

     

2-Deoxyglucose transport by the frog sartorius: Effects of electrical stimulation and N-carbobenzoxy-glycyl-l-phenylalaninamide

• 1.1. Studies characterizing glucose transport in the frog sartorius were performed.
• 2.2. For nonstimulated and stimulated muscles, intracellular 2-deoxyglucose exceeded 2-deoxyglucose-6-phosphate at 15 min, showed little further increase, and was maintained below the extracellular concentration for 2 hr.
• 3.3. Accumulated 2-deoxyglucose-6-phosphate did not inhibit glucose transport.
• 4.4. Unlike in adipocytes, basal and stimulated 2-deoxyglucose transport showed no difference in sensitivity to N-carbobenzoxy-glycyl-l-phenylalaninamide.
• 5.5. Phenylarsine oxide blocked contraction-enhanced 2-deoxyglucose uptake.
• 6.6. These results suggest that the glucose transporter of the sartorius exhibits auto-regulation, and that basal transport is not regulated by the same process as in adipocytes.

     

Induction of a cadmium-binding protein in a unicellular alga

• 1.1. An inducible Cd-binding compound has been detected in a Cd-treated euryhaline alga, Dunaliella bioculata.
• 2.2. The apparent molecular mass of this heat-stable Cd-binding compound, as determined by gel filtration, was about 10,000.
• 3.3. Anion exchange chromatography (on DEAE Sepharose and Mono Q) and autoradiography of non-denaturing PAGE of the M_r 10,000 fractions revealed the presence of a highly acidic Cd-binding protein which differs on its properties from mammal metallothioneins.

     

Osmoregulatory adjustments by three atherinids (Leptatherina presbyteroides; Craterocephalus mugiloides; Leptatherina wallacei) to a range of salinities

• 1.1. Patterns of osmoregulation were studied in three species of Swan river atherinids (Leptatherina presbyteroides, lower estuarine and marine; Craterocephalus mugiloides, mid estuarine; Leptatherina wallacei, upper estuarine) over a wide range of salinities.
• 2.2. The plasma Na⁺ concentration was elevated with an increase in salinity.
• 3.3. Haematocrit and body water content decreased with acclimation to higher salinity.
• 4.4. All three species of atherinids osmotically regulated over a salinity range greater than that which these fish are reported to occur in.

     

Laboratory studies on the oxygen consumption of the marine teleost,Lichia amia (Linnaeus, 1758)

• 1.1. The oxygen consumption of the marine teleost, Lichia amia was investigated under controlled laboratory conditions.
• 2.2. The routine oxygen consumption showed a strong circadian rhythm with the fish being mainly active during the light period.
• 3.3. The specific mass exponent (dimension: μg O₂/g/hr) is temperature independent and ranges from 0.27–0.29.
• 4.4. Starving the fish results in a mean decrease in active, routine and standard oxygen consumption of 21%, 24% and 20%, respectively.
• 5.5. Feecling led to an increase in the oxygen consumption of the teleosts, with the mean metabolic rate over the 24 hr that followed, being 58% and 50% higher for fish that had been starved for 162hr and 40 hr, respectively.
• 6.6. Apparent SDA showed some variation and ranged from 6.0 to 35.5%.
• 7.7. The results obtained are generally in agreement with those recorded for other teleosts.

     

Oxygen binding by the hemocyanin of Busycon canaliculatum

• 1.1. This study examined the effect of the monoamines dopamine and octopamine, as well as tyrosine on the oxygen affinity and cooperativity of oxygen binding by the hemocyanin of the marine gastropod Busycon canaliculatum. The effect of temperature on hemocyanin oxygen affinity was also examined.
• 2.2. Freezing Busycon hemocyanin did not affect the binding of oxygen.
• 3.3. Dopamine, octopamine and tyrosine had no significant effect on the oxygen affinity or cooperativity of oxygen binding by the hemocyanin of B. canaliculatum.
• 4.4. It was concluded that Busycon hemocyanin either has no binding sites for the two monoamines or for tyrosine, or that binding of the molecules has no functional significance.
• 5.5. Both temperature sensitivity and affinity of hemocyanin-oxygen binding were similar to values previously reported for hemocyanin of Busycon from other localities.

     

Distribution of opine dehydrogenases and lactate dehydrogenase activities in marine animals

• 1.1. Opine dehydrogenases (OpDHs) and lactate dehydrogenase (LDH) activities were determined in various marine animals. OpDHs were detected in six marine invertebrate phyla; Porifera, Coelenterata, Annelida, Mollusca, Arthropoda and Echinodermata in phylogenic sequence.
• 2.2. Among several OpDHs, tauropine dehydrogenase (TaDH) occurred widely in marine invertebrates, from Porifera to Echinodermata.
• 3.3. With a few exceptions, total OpDHs activities exceeded that of LDH activity in the marine invertebrates investigated.
• 4.4. With respect to anaerobic glycolysis, OpDHs are indicated to play an important role in phylogenically lower invertebrates, whereas LDH is more important in higher animals.

     

Hyperosmotic properties of the fluids of the perivisceral coelom and water vascular system of starfish kept under stable conditions

• 1.1. Osmotic measurements were made on the perivisceral coelomic and water vascular fluids of 4 species of northwest Pacific starfish and their stable sea-water media.
• 2.2. Mean levels of both fluids were hyperosmotic in every species, often at statistically significant levels.
• 3.3. For all species combined, mean hyperosmolality (mosmol/kg ± SE) of perivisceral coelomic fluid was 1.49 ± 0.17, and water vascular fluid 6.07 ± 0.74.
• 4.4. The hyperosmotic nature of these fluids contributes to water balance, working in conjunction with madreporitic inflow and other factors.

     

Is the metal binding protein metallothionein present in the coelenterate Hydra attenuata?

• 1.1. Studies have been performed on untreated and heavy metal treated Hydra attenuata in order to reveal the presence of low mol. wt metal-binding proteins.
• 2.2. A prepared rat metallothionein (Mt) standard, gel permeation, polyacrylamide gel electrophoresis and autoradiographic techniques were used in the experiments.
• 3.3. Our results indicate that H. attenuata, and three species of marine coelenterates, lack metallothionein (Mt) or other metal binding proteins.

     

Growth and haematological response of sturgeon hybrids Russian sturgeon (Acipenser guldenstadti brandt) × beluga (Huso huso L.) to protein and lipid contents in the diet

• 1.1. Fingerlings of intergenious hybrid Russian sturgeon (Acipenser guldenstadti) × beluga (Huso huso) weighing 22 g reared in water with salinity 18 ppt were fed nine diets differing in protein and fat content.
• 2.2. The increase of dietary protein content (from 45 to 52%) improved the fingerlings growth rate, food and protein conversion efficiencies. No effect of further protein content increase to 60% was observed.
• 3.3. The increase of dietary fat content from 10 to 20% positively influenced all growth results.
• 4.4. The muscular lipid content increased following the increase in dietary fat due to accumulation of triacylglycerols.
• 5.5. Distinctive leucopenia in neutrophils and leucophilia in lymphocytes following dietary protein and fat content increase were observed.
• 6.6. It was concluded that within the analysed range of values the increase of dietary protein and lipid content improved the physiological status of sturgeon hybrid fingerlings.

     

Chemical intermediates in α-methylnoradrenaline oxidation by tyrosinase—I. Spectral properties and stoichiometry

• 1.1. A pathway for a-methylnoradrenaline oxidation to α-methylnoradrenochrome, by tyrosinase, is proposed. Characterization of intermediates in this oxidative reaction and stoichiometry determination have both been performed.
• 2.2. It has been possible to detect spectrophotometrically o-quinone-H⁺ as the first intermediate in this pathway after oxidizing α-methylnoradrenaline with mushroom tyrosinase or sodium periodate in a pH range from 5 to 6.
• 3.3. The steps for α-methylnoradrenaline transformation into its aminochrome could be: α-methylnoradrenaline → o -α-methylnoradrenaline — H⁺ → o — α -methylnoradrenalinequinone → leuko — α — methylnora — drenochrome→α-methylnoradrenochrome.
• 4.4. No participation of oxygen was detected in the conversion of leuko-α-mehtylnoradrenochrome into α -methylnoradrenochrome.
• 5.5. Matrix analysis of the spectra obtained with a rapid scan spetrophotometer verified that o-quinone-H⁺ was transformed into aminochrome in a constant ratio.
• 6.6. The stoichiometry for this conversion followed the equation: 2 α-methylnoradrenalinequinone-H⁺ → α-methylnoradrenaline + α-methylnoradrenochrome.

     

Extraction and rapid identification of law molecular weight compounds from marine organisms

• 1.1. Different methods for extraction of low molecular weight compunds (lipids, terpenoids, steroids, glycosides, etc.) from Black Sea invertebrates and algae were examined.
• 2.2. The composition and quantity of the extracts were found to depend strongly on the type of the organisms and on the solvents used for extraction.
• 3.3. Thin-layer chromatography was found to be convenient for rapid identification of low molecular weight compounds in the extracts.
• 4.4. An approach has been developed which was suitable for field experimentrsand was successfully applied for the analysis of five algae and seven invertebrates from the Black Sea.