Lipid classes and fatty acid composition of rotifers (Brachionus plicatilis) fed two algal diets

Rotifers (Brachionus plicatilis), maintained on baker's yeast, were fed for 24h upon two algal diets, Isochrysis galbana (diet A) and Isochrysis galbana + Nannochloropsis gaditana (diet B). (These algal diets were selected for their potential use as essential fatty acid (EFA) boosters, taking into account the requirements of fish larvae). The effect of these algal diets on total lipid content, lipid classes and fatty acid composition was studied. The total lipid content increased after feeding upon both diets but no significant differences were found between the two types. Neutral lipid and polar lipid contents increased and a positive correlation was observed between the neutral lipids content of rotifers and that of the food supplied. However, the content of polar lipids in rotifers did not depend upon that of the diet. The increase in neutral lipid content was found to be higher in rotifers fed upon diet B, compared to diet A which increased the phospholipid content. Non-enriched rotifers contained only small amounts of polyenoic fatty acids, i.e. 18:3n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3, the contents of which increased significantly by feeding both diets. The EFA composition (20:4n-6, 20:5n-3 and 22:6n-3) of neutral lipids and phopholipids in rotifers reflected the EFA composition of each diet. Diet B-fed rotifers had the highest content in 20:4n-6 and 20:5n-3, whereas rotifers fed diet A and the highest 22:6n-3 content. The mixed diet I. galbana + N. gaditana enhanced substantially the composition of lipid classes i.e. neutral lipids and of n-3 PUFA of rotifers in comparison with Isochrysis or yeast diets.     

Glutamic acid decarboxylase (GAD)-like immunoreactivity in the pedal ganglion of Mytilus galloprovincialis

Summary A substance immunologically related to vertebrate glutamic acid decarboxylase (GAD) has been visualized in the pedal ganglion of Mytilus with the pre-embedding peroxidase-antiperoxidase method, by use of an antiserum raised in sheep against rat brain GAD. The results show that GAD-like immunoreactivity is present both in neuronal perikarya and in nerve fibers. Positive neurons are located mainly among the fibers of the ganglion neuropil at the commissural level, and more rarely close to unreactive cortical cell bodies. Immunoreactive nerve fibers are observed throughout the neuropil and also in cerebropedal and pedal nerves.Supported by Ministero Pubblica Istruzione (40%)     

Effect of cobalt ions on the metabolism of some volatile and polar compounds in the marine invertebrates Mytilus galloprovincialis and Actinia equina

The compositions of the volatile and polar fractions from two coexisting Black Sea invertebrates, the mussel Mytilus galloprovincialis and the beadlet anemone Actinia equina, were established. The main metabolites in the volatile fraction from the investigated animals appeared to be methyl esters of fatty acids and fatty aldehydes. In the polar fraction from both animals low concentrations of free acids and nitrogen-containing compounds were obtained. Free carbohydrates were in much higher concentrations in M. galloprovincialis than in A. equina. Some sterols, probably as polar conjugates, were identified mainly in A. equina. Significant changes among all compounds appeared after treatment of both invertebrates with two different concentrations of cobalt ions. The variety of changes in each invertebrate could be due to their different evolutionary status. The effect of cobalt ions was often stronger at medium cobalt-ion concentrations.     

The HOX Gene Cluster in the Bivalve Mollusc Mytilus galloprovincialis

The clustered Hox genes play a central role in the regulation of development in bilaterian animals. In this study, we analyzed the homeobox-containing genes in a bivalve mollusc, the mussel Mytilus galloprovincialis, an unsegmented spiralian lophotrochozoan. We isolated and characterized four Hox cluster genes using the polymerase chain reaction with specific primers. Molecular alignments and phylogenetic analysis indicate that these mussel genes are homologs of the anterior group (pb ortholog), paralog group 3, and central group (PG4/Dfd and PG5/Scr) genes. The putative homeodomain sequences were designated Mgox1, Mgox2, Mgox3, and Mgox4.     

Artificial diets determine fatty acid composition in edible Ruspolia differens (Orthoptera: Tettigoniidae)

There are increasing interests in rearing edible insects in Africa, but information on how the feeds modify their fatty acids is largely lacking. In this work, the influence of artificial diets on the fatty acid contents and composition in the edible Ruspolia differens (Serville, 1838), in Uganda was assessed. R. differens was reared on the mixtures of six gradually diversified diets of two, three, four, six, eight and nine feeds. The diets were formulated from rice seed head, finger millet seed head, wheat bran, superfeed chicken egg booster, sorghum seed head, germinated finger millet, simsim cake, crushed dog biscuit pellet and shea butter. Fatty acid methyl esters were prepared using direct transesterification method, and analysed using gas chromatography. The contents of saturated, monounsaturated and polyunsaturated fatty acid differed significantly among the diets. The more diverse diets resulted in increased content of the polyunsaturated fatty acids. The n6:n3 ratio differed significantly among the diets and between the sexes, with R. differens fed on the four-feed diet having a higher n6:n3 ratio than those fed on other diets. Also, the fatty acid composition differed significantly among the diets, and diet diversification corresponded with the proportions of polyunsaturated fatty acids, especially linoleic acid. Overall, our results demonstrate that higher levels of essential fatty acids can be achieved by rearing R. differens on highly diversified diets. These findings are important in informing the design of future mass-rearing program for this edible insect.     

The metallothionein genes of Mytilus galloprovincialis: genomic organization, tissue expression and evolution

Recently, increasing interest has been directed to the study of metallothioneins (MTs), which are small proteins that are able to bind metal ions. The induction of MT synthesis after exposure to metal or other environmental contaminants in a large number of aquatic invertebrates makes these proteins good biomarkers in water monitoring programs. Within bivalves, the species Mytilus galloprovincialis and Mytilus edulis represent model organisms for these types of studies, as well as for molecular studies regarding the expression and characterization of MT encoding genes. In the present paper, we focused on the genomic characterization, evolutionary, and tissue-expression analyses of the MT-10, MT-10 Intronless, and MT-20 genes in M. galloprovincialis. The comparison of the genomic sequences showed the presence of long nucleotide stretches within the introns of the MT genes that are conserved between M. galloprovincialis and M. edulis. These non-coding conserved sequences may contain regulatory motifs. Real-Time RT-PCR experiments revealed that, at the basal conditions, the MT-10 and MT-10 Intronless genes are expressed at levels considerably higher than the MT-20 gene, mainly in the digestive gland and gill tissue. The strong induction of the MT-20 gene expression detected in a field-collected sample is associated with the up-regulation of both the MT-10 and MT-10 Intronless genes. Evolutionary analysis revealed signals of localized positive selection that, together with the tissue-expression data, support a possible functional diversification between the MTs encoded by the MT-10 and MT-10 Intronless genes.     

Changes in fatty acid composition in Pseudomonas putida and Pseudomonas stutzeri during naphthalene degradation

The effects of naphthalene on the whole cell-derived fatty acid composition of Pseudomonas putida and Pseudomonas stutzeri during naphthalene degradation were investigated. These strains differed in their abilities to degrade naphthalene and in 1,2-catechol dioxygenase activities. The cells of both strains reacted to the addition of naphthalene with an increase in the saturated/unsaturated ratio. The dynamic changes comprised also alterations in the percentage of hydroxy, cyclopropane and branched fatty acids. Upon the exposure of naphthalene, new fatty acids were detected.     

Distribution of 5HT-immunoreactivity in the pedal ganglion of Mytilus galloprovincialis

Summary Serotonergic cell bodies and fibers were identified in the pedal ganglia of Mytilus gattoprovincialis with a serum raised against serotonin and the unlabelled peroxidase-antiperoxidase pre- and post-embedding methods. Examination of Vibratome and serial semithin sections showed that most reactive perikarya are located in the ganglionic cortex, being mainly concentrated at the medial aspect of the postero-dorsal portion of the ganglia. Immunoreactive fibers form a dense network in the neuropil, extend throughout the commissure and run parallel in the nerves and connective tracts. The morphology of serotonin-positive cells compared with that of Golgi-impregnated neurons allows the identification of a main population of unipolar, probably projecting neurons and of smaller multipolar cells likely representing local circuit elements. The ultrastructure of labelled neurons is comparable to that of serotonergic cells described in both vertebrate and invertebrate nervous systems.Supported by Ministero Pubblica Istruzione (40%)     

Changes in the Fatty Acid Composition of Hepatopancreas of the Mollusk Mytilus trossulus Fed on Microalgae

The influence of diet on the fatty acid composition of the hepatopancreas of Mytilus trossulus was studied. Three groups of mollusks were fed monocultures of the microalgae Phaeodactylum tricornutum, Chaetoceros muelleri (Bacillariophyceae), and Nannochloropsis sp. (Eustigmatophyceae) for 10 days. After 10 days, the proportion of polyunsaturated fatty acids, mainly eicosapentaenoic and docosahexaenoic, increased in the total lipids of the hepatopancreas in all mollusk groups. The content of saturated fatty acids in the mussel tissues decreased and was not dependent on the amount in the algal diet. Toward the end of the experiment, the fatty acid composition of the hepatopancreas of mussels was similar irrespective of the fatty acid composition of their food. The fatty acid analysis of M. trossulus feces suggests a selective assimilation by mussels of predominantly the n-3 polyunsaturated fatty acids. The role of fatty acid metabolism in M. trossulus is discussed.     

半滑舌鳎受精卵、卵黄囊仔鱼和开口仔鱼氨基酸及脂肪酸的变化

采用气相色谱仪和氨基酸分析仪测定了半滑舌鳎(Cynoglossus semilaevis)受精卵、卵黄囊仔鱼和开口仔鱼的氨基酸与脂肪酸组成的变化。结果表明:总氨基酸组成在受精卵和卵黄囊仔鱼之间变化明显,但是在卵黄囊仔鱼和开口仔鱼之间只有细微的变化。开口仔鱼与其摄食的轮虫的总必需氨基酸组成相关。受精卵、卵黄囊仔鱼、开口仔鱼的游离氨基酸含量分别为139 mg/g、3.6 mg/g和2.5 mg/g,占总氨基酸含量的22.3%、3.6%和2.5%。饱和脂肪酸的总量从受精卵到卵黄囊仔鱼明显下降,但是发育到开口仔鱼含量无显著变化。单不饱和脂肪酸和多不饱和脂肪酸的总量在不同发育阶段无显著变化,而EPA和DHA的含量从卵黄囊仔鱼到开口仔鱼有明显下降。这表明在早期发育阶段半滑舌鳎主要利用饱和脂肪酸作为能量代谢的基质,对饱和脂肪酸的利用程度大于单不饱和脂肪酸和多不饱和脂肪酸。半滑舌鳎似乎需要长链的多不饱和脂肪酸如EPA、DHA和ARA。     

Different fatty acid composition in central and peripheral adipose tissues of the American mink (Mustela vison)

Fatty acid (FA) composition in the intraabdominal (IAB), subcutaneous (SC) and peripheral adipose tissues of the semiaquatic American mink (Mustela vison) was examined in comparison to the diet by gas-liquid chromatography. There was a clear compositional gradient from the IAB via SC to peripheral adipose tissues and the anatomically different adipose tissues accumulated or metabolized FA selectively. The total lipids of the body appendages had smaller proportions of saturated (SFA) and larger proportions of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) than the lipids of the trunk adipose tissues. Especially n-3 PUFA were enriched in the periphery. The appendages were also characterized with a high ratio of unsaturated FA to SFA, an increased Delta9-desaturation index and increased mean numbers of double bonds and carbon atoms in a FA molecule. The proportions of SFA and MUFA of the diet resembled the trunk adipose tissues while the dietary percentage of n-3 PUFA surpassed those of the trunk fat depots but was lower than those of the peripheral fats. These data confirm that the FA signatures of mammals reflect not only their dietary history but also metabolic modifications of ingested FA.     

Effect of growth phase on the fatty acid composition of Thermus spp.

Two yellow and two red pigmented strains of Thermus were monitored for changes in fatty acid content and composition with reference to growth phase at the optimum temperature. Fatty acid content per mg of dry weight increased as the cultures aged. In addition the quantities of iso C 15:0, iso C 17:0 and iso C 16:0 increased in yellow pigmented strains, but in red pigmented strains, an increase was seen in iso C 15:0, but C 16:0 and iso C 16:0 levels decreased. Thus the fatty acid composition of these organisms varies with growth phase, and shows also strain specific variability.     

Seasonal variations of arsenic in mussels<Emphasis Type="Italic"> Mytilus galloprovincialis</Emphasis>

Total arsenic concentration in the edible part of mussels Mytilus galloprovincialis was evaluated seasonally in the coastal area of Rijeka Bay (North Adriatic Sea, Croatia). Sampling stations were located close to the City of Bakar with no industrial facilities (site 1), in the vicinity of the oil refinery and oil thermoelectric power plant (Urinj, site 2), and 4 miles away from the Plomin coal thermoelectric power plant (Brse village, site 3). Additionally, the concentration of arsenic in the tail muscle of the lobster Nephrops norvegicus, collected in Rijeka Bay, was studied. During winter at sites 2 and 3, the total arsenic in the edible part of the mussels was 16.4 mg As/kg FW (FW=fresh weight) and 4.38 mg As/kg FW, respectively, and increased during springtime at site 2 (6.5 mg As/kg FW) compared to the rest of the year, when individual total arsenic concentration at all sites ranged from 1.7 to 3.7 mg As/kg FW. In the winter (sites 2 and 3) and springtime (site 2) there was no correlation between the length of the mussel shell and the arsenic concentration in the edible part of the mussels. In the other seasons, at sites 1, 2 and 3, there was a correlation between arsenic in the edible part of mussels and shell length in most cases (correlation coefficients r varied from 0.64 to 0.85; P <0.05 to P <0.01). Correlation between shell length (in the narrow range of shell lengths from 3.4 to 5.0 cm) and arsenic in the edible part of the mussels shows linearity with a high regression coefficient ( r =0.914; P <0.001). The increase of arsenic in the mussels during winter and spring was suggested at least partially as a result of a low nutritional status, i.e. reduced weight of the mussels edible part during winter. In addition, a linear relationship was found between body length and arsenic concentration in the tail muscle (mean 17.11±4.48 mg As/kg FW) of the Norway lobster.Communicated by H.-D. Franke     

Effect of salinity of medium on cellular fatty acid composition of marine algaPorphyridium cruentum (Rhodophyceae)

Porphyridium cruentum Näg. (clone 161) was found to grow best in medium containing between 0.45 M and 0.8 M NaCl. From studies done on growing cultures, the palmitic acid content of the cells decreased with increasing NaCl concentration of the medium. Conversely, when the culture was transferred from a 0.8 M NaCl medium to 0.2 M NaCl, the amount of palmitic acid in thePorphyridium cells increased with time of incubation and it contributed up to 64.5% of the total fatty acid content. There appears to be a negative correlation between the cellular content of palmitic acid and the growth lag. The oleic acid content varied only marginally with increasing NaCl concentration. The poly-unsaturated acid content (linolenic and arachidonic acids) decreased initially and then increased with NaCl concentration up to and beyond ca. 0.8 M NaCl respectively. At 1.5 M NaCl, the poly-unsaturated fatty acids amounted to 78.2% of the total fatty acids in the cell. For stationary phaseP. cruentum cultures, a similar relationship existed between fatty acids and NaCl concentration. However, palmitic acid was accumulated up to three-fold more when compared to the exponential culture grown in low salinity. In addition stearic acid was also found in significant quantities.     

Effect of choline chloride on fatty acid chain ordering in membranes of wheat (Triticum aestvium L. cv. Miranovskaja 808)

A winter variety (Miranovskaja 808) of wheat (Triticum aestivum L.) was grown in water cultures containing 0, 5, 15, 30, and 60 mM choline chloride. There was an increase in the phosphatidylcholine level at the expense of phosphatidic acid, parallel with an increasing choline concentration in the cultivating medium. While the ratio of free sterols to phospholipids remained essentially constant in the seedlings, there was an increase in the ratio of saturated-to-unsaturated fatty acids of the phospholipid fraction. Probing the protoplasts obtained from the leaves of the seedlings, with spin labeled 5-doxyl- and 16-doxyl stearic acids, indicated a progressive rigidifying of the hydrophobic core of the plasmalemma. It is suggested that this is a manifestation of compensatory mechanisms by which plants attempt to maintain unchanged average membrane fluidity across their membranes in response to the fluidizing effect of choline head groups.     

Changing fatty acid composition during somatic embryogenesis in cultures of Daucus carota

The fatty acid composition of five embryo stages following the induction of embryogenesis in cultures of Daucus carota have been studied. Quantitative rather than qualitative changes in fatty acid content were observed, although globular embryos do possess some fatty acids of long (C 20 and above) chain length not observed in earlier or later stages in development. The major fatty acids present were C 16:0, 18:0, 18:1, 18:2 and 18:3.Abbreviations SM small meristematic - LV large vacuolated - GLC gas-liquid chromatography - TLC thin lager chromatography     

Influence of culture temperature on the growth,biochemical composition and fatty acid profiles of six Antarctic microalgae

The growth, biochemical composition and fatty acid profiles of six Antarctic microalgae cultured at different temperatures, ranging from 4, 6, 9, 14, 20 to 30 ^∘C, were compared. The algae were isolated from seawater, freshwater, soil and snow samples collected during our recent expeditions to Casey, Antarctica, and are currently deposited in the University of Malaya Algae Culture Collection (UMACC). The algae chosen for the study were Chlamydomonas UMACC 229, Chlorella UMACC 234, Chlorella UMACC 237, Klebsormidium UMACC 227, Navicula UMACC 231 and Stichococcus UMACC 238. All the isolates could grow at temperatures up to 20 ^∘C; three isolates, namely Navicula UMACC 231 and the two Chlorella isolates (UMACC 234 and UMACC 237) grew even at 30 ^∘C. Both Chlorella UMACC 234 and Stichococcus UMACC 238 had broad optimal temperatures for growth, ranging from 6 to 20 ^∘C (μ = 0.19 – 0.22 day^–1) and 4 to 14 ^∘C (μ = 0.13 – 0.16 day^–1), respectively. In contrast, optimal growth temperatures for NaviculaUMACC 231 and Chlamydomonas UMACC 229 were 4 ^∘C (μ = 0.34 day^–1) and 6–9 ^∘C (μ = 0.39 – 0.40 day^–1), respectively. The protein content of the Antarctic algae was markedly affected by culture temperature. All except Navicula UMACC 231 and Stichococcus UMACC 238 contained higher amount of proteins when grown at low temperatures (6–9 ^∘C). The percentage of PUFA, especially 20:5 in Navicula UMACC 231 decreased with increasing culture temperature. However, the percentages of unsaturated fatty acids did not show consistent trend with culture temperature for the other algae studied.     

Identification of the steroid fatty acid ester conjugates formed in vivo in Mytilus edulis as a result of exposure to estrogens

Vertebrate-type sex steroids have been detected in a number of mollusk species and may play a role in the reproductive physiology of the animal. Mollusks are also exposed to exogenous estrogenic steroids that are present in sewage effluents, and these may add to the estrogenic burden of exposed animals. We investigated the uptake of estrogens in the blue mussel, Mytlius edulis and report for the first time the identity of estrogen fatty acid ester metabolites formed in vivo in an invertebrate. We exposed mussels to waterborne radiolabeled [(14)C]-17beta-estradiol (E2) or estrone (E1) and determined the nature of their metabolites using radio-HPLC and mass spectrometry (MS). After 13 days of exposure to 10ng/L E2, concentrations of radiolabeled residues were 2428-fold higher in M. edulis soft tissues compared with the ambient water concentration of E2. All the E2 residues in the mussel were present as a lipophilic ester which, in depuration studies, had a half-life of 8.3 days. Exposure of mussels to [(14)C]-E1 (70ng/L) resulted in formation of a similar lipophilic metabolite that after hydrolysis released [(14)C]-E2. Tandem MSMS analyses of the purified steroid ester fraction isolated from mussels exposed to either E2 or E1 revealed that they had the same composition and comprised C16:0, C16:1 and C16:2 esters of E2. This work reveals that in vivo E1 is rapidly metabolized to E2 in mussels prior to conjugation to C16 fatty acid esters, proving that C17-ketoreductase and C16 fatty acid acyl-CoA:E2 acyltransferase are important enzymes for the metabolism of estrogens in M. edulis.