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morphology and ultrastructure of cryptobia eilatica n. sp. (bodonidae: kinetoplastida), an ectoparasite from the gills of marine fish.a marine kinetoplastid flagellate, cryptobia eilatica n. sp., is described from the gills of cultured gilt-head sea bream sparus aurata l. and wild black-spot sea bream diplodus noct (valenciennes) in the red sea. the trophozoite is elongated and lacks a contractile vacuole and undulating membrane. the body averages 13.5 x 4.1 microns, anterior flagellum 9.7 microns and free portion of recurrent flagellum 15.2 microns. the ultrastructural features of the species exhibit great similarity to vario ...19902086780
extraction of lipids and lipophilic antioxidants from fish tissues: a comparison among different methods.1. lipids, phospholipids and lipid soluble antioxidants were extracted from sparus auratus liver and white muscle by three different methods and the yields obtained were compared. 2. none of the three procedures can recover the above components with the same efficiency. 3. for comparison the methods were also applied to rat liver homogenates. 4. the choice of the extraction procedures depends on the tissue investigated and on specific research requirements.19921582177
immunocytochemistry of somatotrophs, gonadotrophs, prolactin and adrenocorticotropin cells in larval sea bream (sparus auratus) pituitaries.the chronological appearance of endocrine cells in the pituitary of sea-bream (sparus auratus) larvae was studied using antisera against salmon prolactin, trout growth hormone, salmon gonadotropin and n-terminal human adrenocorticotropin. the larval pituitary (1-12 days after hatching) was oval in shape and was composed of a dense mass of cells with few neurohypophysial fibres. by 60 days after hatching it began to resemble the adult and was divisible into a distinct rostral pars distalis contai ...19921330314
[study of the larval stages of monogenea of fishes (author's transl)].the ciliate cells and the oncomiracidial chetotaxy of the following species are described: among the polyopisthocotylea : gastrocotyle trachurii van bneden and hesse, 1863, gastrocotylidae, parasite of trachurus trachurus (l.); cyclocotyla bellones (otto, 1821), diclidophoradae, living on the cymothoides isopods of sparidae : boops boops (l.) and spicara maena (l.) ; mazocraes sp., mazocraeidae, parasite of sardinella maderensis (lowe, 1841). among the monopisthocotylea : ligophorus vanbenedeni ...1978754614
fructose 2,6-bisphosphate in liver of sparus aurata: influence of nutritional state.1. fructose 2,6-bisphosphate (fru-2,6-p2) has been measured in liver and muscle of gilthead sea bream fish, sparus aurata. 2. the fru-2,6-p2 levels in liver depend on the diet given to the fish: in fish fed a high carbohydrate diet, the fru-2,6-p2 levels are higher than any one previously reported. these changes are associated with differences in the phosphofructokinase 2 activity. 3. fru-2,6-p2 levels has also been measured in liver of sparus aurata after different fasting periods. in starved f ...19902142067
evidence for a direct action of gh on haemopoietic cells of a marine fish, the gilthead sea bream (sparus aurata).receptors for gh were characterized in the head kidney of gilthead sea bream (sparus aurata), using radioiodinated and biotinylated ligands. the specific binding of radiolabelled recombinant gilthead sea bream gh (rsbgh) to head kidney membrane preparations was dependent on membrane concentration. salmon prolactin, salmon gonadotrophin and carp gonadotrophin did not compete for 125i-labelled rsbgh-binding sites. unlabelled rsbgh competitively displaced 125i-labelled rsbgh bound to head kidney me ...19957595141
cloning and characterisation of a fish aldolase b gene.a full length cdna clone representing an aldolase mrna was isolated from a sea bream (sparus aurata) liver cdna library. sequencing of this clone revealed it to encode a 364 amino acid protein with 74% amino acid identity to human aldolase b and slightly lower similarity to human aldolase a and c. in view of the sequence data and of northern blot analysis showing strong expression of a 1.6 kb transcript in liver it was concluded that the cloned gene represents aldolase b. this clone represents t ...19957632737
the muscle isoform of 6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase of the teleost sparus aurata: relationship with the liver isoform.the liver isoform of 6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase of the teleost fish sparaus aurata has several characteristics similar to the skeletal muscle isoform of mammals. in order to ascertain the relation between muscle and liver isoforms in teleost, 6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase was purified from skeletal muscle of s. aurata. the muscle isozyme is composed of subunits with a molecular weight of 54 kda, is bifunctional, and has an activity ratio kinase to ...19957646054
degradation of gonadotropin-releasing hormones in the gilthead seabream, sparus aurata. i. cleavage of native salmon gnrh and mammalian lhrh in the pituitary.the pattern and kinetics of degradation of native salmon gonadotropin-releasing hormone (sgnrh) and mammalian leuteinizing hormone-releasing hormone (lhrh) by pituitary bound enzymes were studied in the gilthead seabream, sparus aurata. sgnrh and lhrh were incubated for different periods of time with membrane or cytosolic fractions of pituitary homogenates. at the end of the incubation, the degradation mixture was fractionated on reverse-phase high-pressure liquid chromatography. the degradation ...19902202610
histochemistry of glycoconjugates in the gallbladder epithelium of ten animal species.a battery of seven lectins and several conventional mucin histochemical techniques were used to identify the epithelial mucins of the gallbladder of ten species: man, rabbit (oryctolagus cuniculus, mammalia), hamster (mesocricetus auratus, mammalia), chicken (gallus gallus, bird), sparrow (passer domesticus, bird), moorish gecko (tarentola mauritanica, reptilia), ladder snake (elaphe scalaris, reptilia), lake frog (rana perezi, amphibia), natterjack toad (bufo calamita, amphibia) and gilthead se ...19892722572
molecular cloning and characterization of a novel gonadotropin-releasing hormone from the gilthead seabream (sparus aurata).a novel form of gonadotropin-releasing hormone (gnrh) had been recently isolated and characterized from the gilthead seabream (sparus aurata). this novel form, designated sbgnrh, was suggested to be the endogenous stimulator of gonadotropin release. this paper reports on the isolation and characterization of a 360-bp cdna encoding the complete sbgnrh precursor. this precursor is composed of a 25 amino acid leader sequence, the biologically active sbgnrh, the cleavage site (gly-lys-arg), and a 57 ...19957749463
gonadotropin-releasing activities of the three native forms of gonadotropin-releasing hormone present in the brain of gilthead seabream, sparus aurata.three forms of gonadotropin-releasing hormone (gnrh) have been recently identified in the brain of gilthead seabream (sparus aurata): salmon gnrh (sgnrh), chicken gnrh-ii (cgnrh-ii), and a novel form, ser8-mammalian gnrh, named seabream gnrh (sbgnrh). sbgnrh is the most abundant form in the pituitaries of sexually mature seabream during the spawning season. the present study investigated the gonadotropin-releasing activities of the three native forms of gnrh found in seabream brains, as well as ...19957789744
growth hormone increases plasma levels of insulin-like growth factor (igf-i) in a teleost, the gilthead seabream (sparus aurata).a heterologous radioimmunoassay (ria) was applied for the determination of immunoreactive (ir)--insulin-like growth factor (igf-i) in a teleost, the gilthead seabream (sparus aurata). serial dilutions of the fish plasma gave a linear curve when added to constant amounts of 125i-labelled human igf-i(53-70) and antiserum prepared against this fragment. the ria was used to study the effect of gh on plasma levels of ir-igf-i in s. aurata. a single injection of human recombinant gh (1 micrograms/g) r ...19892926294
metabolic activation of carcinogenic aromatic amines by fish exposed to environmental pollutants.activation of arylamines to mutagenic metabolites by hepatic s9 fractions has been evaluated as a biomaker of fish exposure to pollutants, using gilthead seabream (sparus aurata), a valuable fish species from the spanish south atlantic littoral, as model organism. to obtain maximal sensitivity to the mutagenic action of aromatic amines, a strain of salmonella typhimurium overproducing o-acetyltransferase was used. fish were treated with aroclor 1254, pesticides (malathion and dieldrin), or coppe ...19957875126
the use of recombinant gilthead sea bream (sparus aurata) growth hormone for radioiodination and standard preparation in radioimmunoassay.a gilthead sea bream growth hormone (sbgh) obtained by cloning and expression of sbgh cdna was used to develop a sensitive and specific radioimmunoassay (ria). iodination of recombinant sbgh (rsbgh) was performed by the classical chloramine-t method. specific antiserum, raised in rabbits, was added in a final dilution of 1/36,000. the minimum detectable dose was 30 pg, and the midrange of the assay (ed50) was 275 pg. intra- and inter-assay coefficients of variation (cv) were 3.3 and 5.8% at ed50 ...19957669108
promutagen activation by fish liver as a biomarker of littoral pollution.metabolic activation of known promutagens by liver s9 fractions of mugil sp. (grey mullet) from two zones of the south atlantic spanish littoral was determined and related to their pollution levels. sediments from the putative contaminated area contained high concentrations of pahs, pcbs and pesticides, and animals from the polluted site exhibited higher concentrations of metals than those from the reference area. hepatic s9 fractions of mullets from the polluted site showed 5.1-, 18.6- and 42.8 ...19947925325
natural hemolytic and bactericidal activities of sea bream sparus aurata serum are effected by the alternative complement pathway.sea bream serum displayed bactericidal and hemolytic activities. these activities were depleted when serum was incubated with different activators of the alternative complement pathway (acp). ethylenediaminetetraacetic acid (edta) inhibited both the hemolytic and bactericidal activities, while ethyleneglycol-bis (b-aminoethyl ether)-n, n, n'-tetraacetic acid (egta) was not inhibitory. an antibody against the putative third component of sea bream component (c3) was produced. it was observed by im ...19957676614
cloning, expression, and characterization of a recombinant gilthead seabream growth hormone.cdna clones coding for the gilthead seabream (sparus aurata) growth hormone (sbgh) were isolated from a pituitary expression library using a flounder cdna probe. the nucleotide sequence of a gh cdna clone containing an insert of 896 nucleotides was determined. the cdna encoded a polypeptide of 204 amino acids including a signal peptide of 17 amino acids and contained a 5' and a 3' untranslated region of 48 and 233 nucleotides, respectively. the mrna determined by northern blot was approximately ...19947851717
non-specific cytotoxic response against tumor target cells mediated by leucocytes from seawater teleosts, sparus aurata and dicentrarchus labrax: an ultrastructural study.the in vitro cytotoxic reaction of leucocytes from the seawater teleosts gilthead seabream (sparus aurata) and sea bass (dicentrarchus labrax) against tumor target cells was studied by transmission and scanning electron microscopy. head-kidney, blood or peritoneal exudate leucocytes were incubated with hela or b16 melanoma cells. after incubation, conjugates of leucocytes (effectors) and tumor cells (targets) were observed. both the effector-target cell binding and target cell lysis steps of the ...19947880589
ovarian melanotropic peptides and adaptation in two teleostean species: sparus aurata l. and dicentrarchus labrax l.the ovarian tissue of dicentrarchus labrax and sparus aurata displays two immunoreactive peaks that correspond to the elution time of human des-acetyl alpha-msh [acth(1-13)-amide] and human alpha-msh. in view of the close identity between the primary structure of fish and human alpha-msh, these data demonstrate that two msh-related peptides are present both in sea bream and sea bass ovary. alpha-msh-like immunoreactivity was found within both granulosa and thecal layers of mature follicles, as w ...19947984515
effects of in vivo benzo(a)pyrene treatment on liver microsomal mixed-function oxidase activities of gilthead seabream (sparus aurata).benzo(a)pyrene [b(a)p] treatment of gilthead seabream, 25 mg/kg, i.p. for 5 consecutive days, did not cause any significant changes in ethylmorphine n-demethylase and aniline 4-hydroxylase activities of liver microsomes. the same treatment did not alter the liver microsomal cytochrome b5 content, nadh-cytochrome b5 reductase and nadph-cytochrome p450 reductase activities. however, benzo(a)pyrene treatment caused a 2-3-fold increase in 7-ethoxyresorufin o-deethylase (7-erod) activity of gilthead ...19948061947
6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase in liver of the teleost sparus aurata.6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase was purified from the liver of the teleost fish sparus aurata and the enzymatic activities were characterized kinetically. both activities copurify, being dimers of relative molecular mass of 98 kda with subunits of m(r) 54 kda. although both specific activities are in the range of mammalian liver isozymes, the kmfru 6-p of teleost 6-phosphofructo 2-kinase is 3 times that in rat liver. the s. aurata 6-phosphofructo 2-kinase is inhibited by adp ...19948109976
three forms of gonadotropin-releasing hormone characterized from brains of one species.most vertebrate species have more than one form of gonadotropin-releasing hormone (gnrh) in their brains, but it is not clear whether each form has a distinct function. we report that sea bream (sparus aurata) brains have three forms of gnrh, one of which is described herein and is called sea bream gnrh (sbgnrh). the primary structures of two forms were determined by edman degradation and mass spectral analysis. the amino acid sequence of sbgnrh is pglu-his-trp-ser-tyr-gly-leu-ser-pro-gly-nh2. t ...19947991588
insulin binding and receptor tyrosine kinase activity in skeletal muscle of carnivorous and omnivorous fish.we characterized the insulin receptors in skeletal muscle from several fish species with different nutritional preferences: brown trout (salmo trutta fario), gilthead sea bream (sparus aurata), tilapia (tilapia mossambica), and carp (cyprinus carpio), semipurified by affinity chromatography (wheat germ agglutinin-agarose). total specific binding and number of receptors per unit weight of piscine white skeletal muscle were lower than those values found in mammalian skeletal muscle. the same param ...19948024051
in vivo effects of the anesthetic, benzocaine, on liver microsomal cytochrome p450 and mixed-function oxidase activities of gilthead seabream (sparus aurata).gilthead seabreams were exposed to benzocaine, 4-aminobenzoic acid ethyl ester, 57 mg/l in sea water for 3 min, daily, for 2 or 3 consecutive days. the fish were killed 20 hr after the last treatment. benzocaine treatment for 2 or 3 days resulted in 57% and 67% inhibition of liver microsomal aniline 4-hydroxylase and ethylmorphine n-demethylase activities, respectively. the total cytochrome p450 content of fish liver microsomes was unaltered following the 2-day benzocaine treatment. however, add ...19948061946
parathyroid hormone-related protein is a factor in normal fish pituitary.using antibodies to the amino-terminal region of human parathyroid hormone-related protein (pthrp) we have demonstrated pthrp immunoreactivity in pituitaries and plasma of the sea bream (sparus aurata). pituitary cells at two distinct locations contained immunodetectable pthrp; an anterior group in the rostral pars distalis which also contained immunoreactive thyroid stimulating hormone (tsh), and a posterior group lying at the border of the pars intermedia and proximal pars distalis between cel ...19938282170
[effect of environmental salinity on the melanotropic cells of the gilthead bream (sparus aurata l.)].the influence of the environmental salinity on the msh cells of the pars intermedia in the euryhaline teleost sparus aurata has been investigated. control animals stayed in sea water (39/1000 salinity), and experimental fish in brackish water (7/1000 salinity) for two months. for light microscopy, pituitaries were fixed with bouin fluid and embedded in paraffin. for electron microscopy they were fixed with karvnosky and embedded in araldite. sections were stained with histochemical procedures an ...19938397430
ontogeny of growth hormone protein and mrna in the gilthead sea bream sparus aurata.the ontogeny of growth hormone (gh) protein and mrna was studied in the gilthead sea bream sparus aurata by northern blot, reverse transcription-polymerase chain reaction (rt-pcr) and western blot analyses. total rna was prepared from a pool of larvae collected on different days after hatching. when hybridized to sparus aurata gh cdna, gh specific mrna was first seen on day 3 post-hatching. levels increased on subsequent days and reached the highest levels on day 9; thereafter a decrease was not ...19968717446
amino-acid sequence of bone gla protein from the african clawed toad xenopus laevis and the fish sparus aurata.as an initial step in the analysis of bone gla protein (bgp; osteocalcin) function in lower vertebrates, we have developed a simple and rapid method for the isolation of bgp from bone and have applied this to the isolation of bgp from the african clawed toad xenopus laevis and the fish sparus aurata. we have also determined the complete amino-acid sequence of sparus and xenopus bgp, including the identification of the sites of y-carboxylation. since the addition of xenopus and sparus bgp sequenc ...19958567186
role of extracellular products in the pathogenicity of vibrio strains on cultured gilt-head seabream (sparus aurata).the enzymatic activities of extracellular products (ecp) from different pathogenic vibrio strains and their virulence for fish (cultured gilt-head seabream, sparus aurata l.) were tested. ecp were obtained by growing the bacteria on cellophane overlays. cytotoxic effects of ecp on fish cell lines were rapid and strong, provoking mortality of fish after 24 to 72 h of the ecp inoculation. a close relationship between proteolytic activities (gelatinase and caseinase) of the ecp and the presence of ...19958588839
immunochemical detection of parathyroid hormone-related protein in the saccus vasculosus of a teleost fish.using antisera to regions of human parathyroid hormone-related protein (pthrp) the saccus vasculosus (sv) of the sea bream (sparus aurata) has been shown to contain immunoreactive pthrp. by immunohistochemistry (ihc) the epithelial coronet cells in fixed and wax-embedded sv tissue reacted with antisera to the prepro region of human pthrp (-13 to +2), the n-terminus pthrp (1-16), and the midmolecule pthrp (50-69). sodium dodecyl sulfate-polyacrylamide gel electrophoresis of saccus extracts and in ...19968713647
light microscopic description of eimeria sparis sp. nov. and goussia sparis sp. nov. (protozoa:apicomplexa) from sparus aurata l. (pisces: teleostei).two new species of coccidia, eimeria sparis sp. nov. and goussia sparis sp. nov., were found in the intestine of sparus aurata from different culture systems of spain. these are the first coccidian species described from this host. e. sparis is distinguished from other eimeria spp. from perciformes in the size and shape of the sporocysts, which measure 6-9.7 (mean = 7.78, sd = 1.33) x4-6.5 (5.3 +/- 0.92) microns, and in the stieda-like body. mature oocysts, spherical or subspherical, measure 9.4 ...19968740548
kinetics of adhesion of selected fish-pathogenic vibrio strains of skin mucus of gilt-head sea bream (sparus aurata l.).the kinetics of adhesion of vibrio strains isolated from diseased fish to skin mucus of gilt-head sea bream was studied. a modified langmuir adsorption isotherm was calculated, and the results obtained indicate that the strains tested (vibrio alginolyticus dp1he4 and vibrio anguillarum-like dc12r8 and dc12r9) showed a saturation kinetics except for v. alginolyticus (can), which showed a proportional adsorption kinetics. the adhesive capability for skin mucus does not seem to be an essential viru ...19968837420
molecular cloning of fish alcohol dehydrogenase cdna.a cdna encoding a putative alcohol dehydrogenase class iii (adh) was cloned from a cdna library constructed from 7-day larvae rna of the marine teleost sparus aurata. the full length cdna is 1350 nucleotides (nt) long and contains an orf of 1128 nt [encoding 376 amino acid (aa) residues]. identity of 82% was found with human class iii adh (305 of 372 aa compared), and only 62% identity with a fish (cod) adh (234 of 375 aa compared). northern hybridization analysis with the cdna revealed a transc ...19968863743
three forms of gonadotropin-releasing hormone in a perciform fish (sparus aurata): complementary deoxyribonucleic acid characterization and brain localization.three forms of gnrh-salmon (sgnrh), seabream (sb-gnrh), and chicken (cgnrh-ii)-have been described in the gilthead seabream (sparus aurata) brain, and the cdna encoding the sbgnrh precursor was recently isolated. in the present study, the cdnas encoding the sgnrh and cgnrh-ii were isolated and characterized, and the neurons producing the three gnrhs were localized in the seabream brain. fragments of sgnrh and cgnrh-ii cdnas were amplified by polymerase chain reaction and used as probes to isolat ...19968862782
crowding stress induces changes in serum haemolytic and agglutinating activity in the gilthead sea bream sparus aurata.sea bream sparus aurata were subjected to crowding for 3 weeks and the levels of plasma cortisol and glucose were determined in order to ascertain the occurrence of stress. at the same time, selected indicators of the immune response were monitored through blood lymphocyte counting and selected humoral responses such as the haemagglutination activity (ha) of serum towards rabbit erythrocytes (rarbc) and the alternative complement pathway (acp) levels. the results show an initial moderate increas ...19968797287
development of enzyme immunoassays for 3,5,3'-triiodo-l-thyronine and l-thyroxine: time-course studies on the effect of food deprivation on plasma thyroid hormones in two marine teleosts, sea bass (dicentrarchus labrax l.) and sea bream (sparus aurata l.).the effects of short-term food deprivation and photoperiod on plasma thyroid hormone levels of sea bass and sea bream were studied. animals were acclimated under constant photoperiod regime (15l/9d) and feeding times (2 hr after light onset and 2 hr before light offset). time-course studies involved monitoring plasma hormone levels every 4 hr throughout 1.5 24-hr cycles. plasma 3,5, 3'-triiodo-l-thyronine (t3) and l-thyroxine (t4) were assayed using a newly developed competitive enzyme immunoass ...19968812399
cloning and expression of somatolactin, a pituitary hormone related to growth hormone and prolactin from gilthead seabream, sparus aurata.a pituitary hormone, somatolactin (sl), belonging to the gh/prl family, is produced in the intermediate lobe of the teleost pituitary. the function of this protein is uncertain. clones coding for sl were isolated and sequenced from a gilthead seabream pituitary cdna expression library. the nucleotide sequence of the larger cdna isolated was 1.5 kb containing a 0.8-kb 3'-untranslated region and two potential polyadenylation signals (aataaa). the mature polypeptide is composed of 207 amino acids, ...19968954766
presence and activity of compounds with gnrh-like activity in the ovary of seabream sparus aurata.the binding and activity of gonadotropin-releasing hormone (gnrh) were characterized in the mature gilthead seabream (sparus aurata) ovary by use of an analogue of salmon gnrh([d-arg6,trp7,leu8,pro9-n(et)]gnrh, sgnrh-a) as labeled ligand. the binding of 125i-sgnrh-a to the seabream ovarian membrane preparation was saturable, displaceable, reversible, and dependent on time, temperature and tissue concentration. addition of unlabeled s-gnrh-a displaced the radio-ligand in a dose-related manner, in ...19979038998
response of eosinophilic granule cells of gilthead seabream (sparus aurata, teleostei) to bacteria and bacterial products.eosinophilic granule cells in the gills and peritoneal exudate of gilthead seabream (sparus aurata l.) are characterized by the presence of prominent eosinophilic granules in their cytoplasm and are here described for the first time. the oval granules of these cells contain an electron-dense inclusion surrounded by a less dense filamentous matrix and are peroxidase- and acid phosphatase-negative. unlike other granulocytes of gilthead seabream, eosinophilic granule cells do not ingest bacteria in ...19979011398
growth hormone as an in vitro phagocyte-activating factor in the gilthead sea bream (sparus aurata)the stimulatory action of growth hormone on gilthead sea bream phagocyte-enriched cultures was demonstrated in vitro for the first time in a fish species. phagocytes consisted mainly of macrophages, with a small number of neutrophils and eosinophils. macrophages were unequivocally identified by their esterase staining and the lack of myeloperoxidase staining. cells primed with recombinant rainbow trout gh showed clear morphological (light- and scanning electron-microscopic) and functional differ ...19979023084
cloning and sequencing of a full-length sea bream (sparus aurata) beta-actin cdna.a full-length cdna clone encoding beta-actin (beta-actin) was isolated from a sea bream (sparus aurata) liver cdna library. sequencing of this clone reveals an open reading frame encoding a 375 amino acid protein that shares a high degree of conservation to other known actins. the sea bream beta-actin sequence showed 98% identity to carp and human beta-actin and 95% and 94% identity to sea squirt and dictyostelium cytoplasmic actins, respectively.19979226879
molecular cloning of gilthead seabream (sparus aurata) pituitary transcription factor ghf-1/pit-1.we report here the complete nucleotide sequence of a cdna clone encoding sparus aurata ghf-1/pit-1 isolated from an expression library prepared from gilthead seabream pituitary gland poly(a)+ rna. the cdna sequence (saghf-1/pit-1) encodes a protein of 371 amino acids (aa) containing a pou domain (aa 194-343) and a transactivation, sta domain (aa 1-128). northern blot hybridization of pituitary rna detected a single 3.0 kb band and a rat ghf-1/pit-1 antiserum was found to immunoreact with pituita ...19979034317
recombinant gilthead seabream (sparus aurata) insulin-like growth factor-i: subcloning, expression in escherichia coli, purification and characterization.gilthead seabream (sparus aurata) insulin-like growth factor-i (gsigf-i) cdna coding for the mature protein was cloned in a pgem-3z vector, and then transferred into prokaryotic expression vector pet-11a and expressed in escherichia coli bl21(de3) cells upon induction with isopropyl thiogalactoside. the expressed protein contained within the inclusion-body pellet was solubilized in 4.5 m urea, refolded for 24 h at ph 11.3 in the presence of catalytic amounts of cysteine and purified to over 98% ...19979135579
ceratomyxa sparusaurati (protozoa:myxosporea) infections in cultured gilthead sea bream sparus aurata (pisces:teleostei) from spain: aspects of the host-parasite relationship.ceratomyxa sparusaurati infection was studied in gilthead sea bream from different mariculture systems of spain. culture conditions were found to influence the infection dynamics. total prevalences ranged from 0 in a closed system to 59.9% in a mediterranean semi-intensive farm. prevalence was significantly lower in summer in a restricted group from the latter system, but no clear seasonal pattern was observed in the remaining fish groups in any system. a statistically significant dependence bet ...19979211504
edwardsiellosis in wild striped bass from the chesapeake bay.the first epizootic of edwardsiellosis, caused by edwardsiella tarda, is described. the epizootic occurred in the chesapeake bay, maryland (usa) during the summer and autumn of 1994, and affected wild adult striped bass (morone saxatilis). clinical signs included numerous irregular coalescing hemorrhagic ulcers on the body and fins that were distinctly malodorous. internally, the body cavity was filled with abundant yellowish or sanguinous mucoid fluid, and the visceral organs had multiple tiny ...19979249698
diversity of the third form of complement, c3, in fish: functional characterization of five forms of c3 in the diploid fish sparus aurata.we have recently shown that sparus aurata, the gilthead sea bream (a diploid species), similarly to rainbow trout (a quasi-tetraploid species), possesses multiple forms of the third form of complement (c3). in the present study we have evaluated the ability of the gilthead sea bream proteins to function as active c3 molecules. all five c3 isoforms could be fixed covalently to sheep erythrocyte ghosts and were able to bind to various complement-activating surfaces in the presence of mgegta. in th ...19979307040
preovulatory changes in the levels of three gonadotropin-releasing hormone-encoding messenger ribonucleic acids (mrnas), gonadotropin beta-subunit mrnas, plasma gonadotropin, and steroids in the female gilthead seabream, sparus aurata.gilthead seabream females undergo daily cycles of final oocyte maturation (fom), ovulation, and spawning throughout their spawning season. fom consists of lipid droplet and yolk granule coalescence, germinal vesicle (gv) migration, and gv breakdown. plasma maturational gonadotropin (gth-ii) levels fluctuate throughout the day, reaching a peak at 8 h before spawning, when the gv is at the periphery of the oocyte. the preovulatory gth-ii surge is accompanied by an increase in the plasma levels of ...19979369182
piscine (sparus aurata) alpha subunit of the g-protein transducin is homologous to mammalian cone and rod transducin.a novel cdna encoding alpha subunit of the gtp-binding protein, transducin, has been cloned from a marine fish, sparus aurata. the cdna contains an open reading frame of 1050 nt (encoding 350 amino acid residues). a high degree of identity was found with known mammalian transducin proteins of cones (gt2 alpha) or rods (gt1 alpha): human gt2 alpha (80.2%), bovine gt2 alpha (79.3%), mouse tt1 alpha (78.2%), mouse gt2 alpha (78%) and bovine gt1 alpha (77.9%). northern blot analysis of different tis ...19979373680
chemotaxis of pathogenic vibrio strains towards mucus surfaces of gilt-head sea bream (sparus aurata l.).vibrio anguillarum and vibrio alginolyticus exhibited significant adhesion to and chemotactic abilities towards mucus collected from the skin, gills, and intestine of gilt-head sea bream. quadratic polynomial models for chemotaxis designed to estimate the influence of temperature demonstrated a differential bacterial chemotaxis depending of the source of the mucus, with the chemotaxis towards intestinal mucus being the least influenced.19989575135
adrenocortical and adrenomedullary homologs in eight species of adult and developing teleosts: morphology, histology, and immunohistochemistry.morphology, histology, and immunohistochemistry of the adrenocortical and adrenomedullary homologs (adrenal glands) of the following developing and adult teleosts were examined: salmoniformes-oncorhynchus mykiss (rainbow trout), salmo trutta fario (brown trout), coregonus lavaretus (white fish); cyprinodontiformes-gambusia affinis (mosquito fish). perciformes-dicentrarchus labrax (sea bass), sparus aurata (sea bream), diplodus sargus (white bream), oblada melanura (saddled bream). the anatomical ...19979405124
identification and spatial distribution of the mrna encoding the gp49 component of the gilthead sea bream, sparus aurata, egg envelope.a cdna encoding the precursor of one of the major components of gilthead sea bream, sparus aurata, egg envelope has been cloned by reverse transcriptase polymerase chain reaction (rt-pcr) techniques. the clone was isolated starting from total rna extracted from the liver of spawning female fish and estradiol-17 beta-treated male fish. sequence analysis revealed that the cdna encoded a protein of 405 aa corresponding to 49-kda component (termed gp49), a glycoprotein belonging to the n-linked type ...19989406196
floor plate and the subcommissural organ are the source of secretory compounds of related nature: comparative immunocytochemical study.the subcommissural organ of vertebrates secretes glycoproteins into the third ventricle that condense to form reissner's fiber (rf). antibodies raised against the bovine rf-glycoproteins reacted with the floor plate (fp) cells of two teleost (oncorhynchus kisutch, sparus aurata) and two amphibian (xenopus laevis, batrachyla taeniata) species. at the ultrastructural level, the immunoreactivity was confined to secretory granules, mainly concentrated at the apical cell pole. in the rostro-caudal ax ...19989482230
methodological aspects of assessing phagocytosis of vibrio anguillarum by leucocytes of gilthead seabream (sparus aurata l.) by flow cytometry and electron microscopy.in this paper we optimize a flow cytometric method for evaluating the phagocytic activity of leucocytes in gilthead seabream (sparus aurata l.) and characterize the phagocytic cells observed. optimal conditions were established for the fluorescein-labelling and analysis of the bacterium vibrio anguillarum by flow cytometry. head-kidney leucocytes were incubated with the heat-killed fluorescein isothiocyanate (fitc)-labelled bacteria for different periods, during which the kinetics of phagocytosi ...19989634605
pathogenicity of vibrio alginolyticus for cultured gilt-head sea bream (sparus aurata l.).the in vivo and in vitro pathogenic activities of whole cells and extracellular products of vibrio alginolyticus for cultured gilt-head sea bream were evaluated. the 50% lethal doses ranged from 5.4 x 10(4) to 1.0 x 10(6) cfu/g of body weight. the strains examined had the ability to adhere to skin, gill, and intestinal mucus of sea bream and to cultured cells of a chinook salmon embryo cell line. in addition, the in vitro ability of v. alginolyticus to adhere to mucus and skin cells of sea bream ...19989797276
cloning, characterisation and expression of the apolipoprotein a-i gene in the sea bream (sparus aurata).a full length cdna clone representing apolipoprotein a-i was isolated from a sea bream (sparus aurata) liver library. the clone encodes a 261 amino acid protein which shows highest amino acid identity (38%) with salmon apolipoprotein a-i. northern blot analysis showed strong expression of a 1.4 kb transcript in liver with lower expression in intestine. expression of apolipoprotein a-i in intestine was markedly reduced by treatment with triiodothyronine (t3).19989804999
molecular cloning of a cdna encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from liver of sparus aurata: nutritional regulation of enzyme expression.a cdna clone encoding full-length 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (6pf-2-k/fru-2, 6-p2ase) was isolated and sequenced from a sparus aurata liver cdna library. the 2527 bp nucleotide sequence of the cdna contains a 73 bp 5'-untranslated region (5'-utr), an open reading frame that encodes a 469 amino acid protein and 1041 bp at the 3'-utr. the deduced amino acid sequence is the first inferred 6pf-2-k/fru-2, 6-p2ase in fish. the kinase and bisphosphatase domains, where the resi ...199910023046
identification of transthyretin in fish (sparus aurata): cdna cloning and characterisation.transthyretin (ttr) has been proposed to have first evolved in reptiles and is one of the three plasma proteins important in the transport of thyroid hormones in higher vertebrates. a full-length cdna encoding ttr was isolated from a sea bream (sparus aurata) liver cdna library using a homologous ttr cdna probe generated by rt-pcr. comparison of the deduced amino acid sequence of sea bream ttr with other published sequences, revealed an overall identity of 47-54%, although the amino acids in the ...199910218999
different putative neuromodulators are present in the nerves which distribute to the teleost skeletal muscle.the presence of putative neuromodulators in the nerve fibres was investigated in white skeletal muscle of two teleost fish not taxonomically correlated and showing different patterns of innervation (multiple versus focal innervation). cryostat sections of epaxial, hypaxial and adductor mandibulae (am) muscles of sparus aurata and anguilla anguilla were stained histochemically for reduced nicotinamide adenine dinucleotide phosphate (nadph)-diaphorase. other sections were used for indirect immunoh ...19989810486
levels of the native forms of gnrh in the pituitary of the gilthead seabream, sparus aurata, at several characteristic stages of the gonadal cycle.brains of the gilthead seabream, sparus aurata, contain three different forms of gonadotropin-releasing hormone (gnrh): seabream (sb) gnrh, chicken (c) gnrh-ii, and salmon (s) gnrh. in the present study, we developed three specific enzyme-linked-immunosorbent assays (elisa) for sbgnrh, cgnrh-ii, and sgnrh and used them to measure the levels of each gnrh form in the pituitary of male and female seabream at different stages of gametogenesis. the sensitivity was 6 pg/well for the sbgnrh assay, 7 pg ...19989843645
cloning of putative piscine (sparus aurata) transthyretin: developmental expression and tissue distribution.cdna encoding putative transthyretin (prealbumin, ttr) was cloned from liver of the marine fish sparus aurata. the cdna contains an open reading frame of 453 nt, encoding for a ttr precursor of 151 amino acids. the deduced amino acid sequence of s. aurata ttr shows identity of 54, 57.3 and 54.1% with lizard, chicken and rat ttr, respectively. northern blot analysis revealed a ttr transcript of about 700 nt, highly expressed in liver, but also in skin. low expression was detected in 12 other tiss ...199910619398
effect of exogenous dna microinjection on early development response of the seabream (sparus aurata).dna transfer techniques allow genetic manipulation of commercial fish. however, marine species have received little attention because of their difficult zootechnical requirements. the seabream (sparus aurata) has become one of the most important species in the aquaculture of mediterranean countries, and the development of suitable dna transfer procedures represents a main step in its genetic improvement. to assess the response of the seabream to exogenous dna, naturally fertilized eggs were inje ...19989892714
epitheliocystis agents in sea bream sparus aurata: morphological evidence for two distinct chlamydia-like developmental cycles.the morphology of membrane-bound intracellular inclusions, or 'cysts', of epitheliocystis from sea bream sparus aurata is described. inclusions under the light microscope appear either granular or amorphous. granular inclusions do not elicit a proliferative host reaction and contain the 3 distinctive developmental stages of chlamydial organisms: the highly pleomorphic reproductive form or reticulate body, the condensing form or intermediate body and the infective non-dividing rather uniform elem ...199910439904
new methods to use fish cytochrome p4501a to assess marine organic pollutants.a new methodology has been developed to assess cytochrome p4501a expression in two south atlantic spanish fish, guilthead seabream (sparus aurata) and grey mullet (liza aurata), used as pollution bioindicators. degenerate oligos were used to amplify by reverse transcription and pcr (rt-pcr) specific cyp1a cdna sequences, used subsequently to design specific primers to get the full cdna by rapid amplification of cdna ends. a new assay has been developed to quantitate cyp1a expression by rt-pcr in ...200010803549
molecular cloning and expression of ovarian cathepsin d in seabream, sparus aurata.in marine fish producing pelagic eggs, the acquirement of buoyancy by the eggs through the hydration process is a key event of reproduction; moreover, the yolk proteolysis, which leads to buoyancy, seems to affect the fertility and survival of the spawned eggs. recently we demonstrated that cathepsin d is the aspartic protease responsible for this intraoocytic processing of vitellogenin into yolk proteins. in the present study, we isolated, cloned, and sequenced the cdna encoding cathepsin d and ...199910456857
liver insulin-like growth factor-i mrna is not affected by diet composition or ration size but shows diurnal variations in regularly-fed gilthead sea bream (sparus aurata).nutritional regulation of insulin-like growth factor-i (igf-i) mrna was assessed in liver of gilthead sea bream (sparus aurata). as in mammals, starvation lowered the igf-i mrna content, which was recovered by refeeding. however, in contrast to previous observations in rats, neither diet composition nor ration size significantly affected hepatic igf-i mrna. although fish growth depended on the quantity of diet supplied, no relationship was found between growth and liver igf-i mrna levels, a fact ...200010736326
differential splicing of three gonadotropin-releasing hormone transcripts in the ovary of seabream (sparus aurata).previous studies demonstrated the presence of high-affinity gnrh binding sites and compounds with gnrh-like activity in the ovary of seabream, sparus aurata, providing evidence for the role of gnrh as a paracrine/autocrine regulator of ovarian function in this species. in the present study, the expression of three forms of gnrh (salmon, chicken-ii, and seabream) genes in this marine teleost species was demonstrated for the first time. moreover, there is evidence for differential splicing and int ...200010775184
glucokinase gene expression is nutritionally regulated in liver of gilthead sea bream (sparus aurata).glucose intolerance in carnivorous fish has been attributed to the lack of hepatic glucokinase (gk) activity. transcription/translation assay and transient transfection of cos-7 cells with a cdna encoding sparus aurata liver gk showed the functionality of the enzyme in vitro. the endogenous fish hepatic gk had lower affinity for glucose than the rat enzyme. the gk activity values in fed fish were similar to those reported for starved and diabetic rats. in this study, we also addressed the nutrit ...200010978515
effects of injecting chitin particles on the innate immune response of gilthead seabream (sparus aurata l.).to determine the effects of chitin (poly [1-->4]-beta-n-acetyl-d--glucosamine) particles on the innate immune response of gilthead seabream (sparus aurata l.), specimens were injected intravenously or intraperitoneally with the substance. natural haemolytic complement activity, head-kidney leucocyte respiratory burst activity and phagocytic and cytotoxic activities were analysed in vitro 3, 5 or 10 days post-injection. all the immune parameters assayed remained unaffected when chitin was intrave ...200011016588
predictive modeling of the shelf life of fish under nonisothermal conditions.the behavior of the natural microflora of mediterannean gilt-head seabream (sparus aurata) was monitored during aerobic storage at different isothermal conditions from 0 to 15 degrees c. the growth data of pseudomonads, established as the specific spoilage organisms of aerobically stored gilt-head seabream, combined with data from previously published experiments, were used to model the effect of temperature on pseudomonad growth using a belehradek type model. the nominal minimum temperature par ...200111282639
application of a systematic experimental procedure to develop a microbial model for rapid fish shelf life predictions.a systematic experimental procedure for fish shelf-life modelling was used to develop a model for predicting the quality of fish in the chill chain. for this, the growth of the naturally occurring bacteria pseudomonads, shewanella putrefaciens, enterobacteriaceae, lactic acid bacteria and yeasts, on gilt-head seabream (sparus aurata), was studied at temperatures from 0 to 15 degrees c. the results from the microbiological, organoleptical and chemical analysis conducted on naturally contaminated ...200011016607
cloning of the bone gla protein gene from the teleost fish sparus aurata. evidence for overall conservation in gene organization and bone-specific expression from fish to man.bone gla protein (bgp, osteocalcin) is a bone-specific vitamin k-dependent protein which has been intensively studied in mammals. although bgp is the most abundant non-collagenous protein of bone, its mode of action at the molecular level remains unclear. from an evolutionary point of view, the appearance of bgp seems to parallel the appearance of hydroxyapatite-containing bone structures since it has never been found in elasmobranchs, whose skeleton is composed of calcified cartilage. according ...200111404005
cloning of the cdna for sea bream (sparus aurata) parathyroid hormone-related protein.this paper reports cloning of the cdna for sea bream (sparus aurata) parathyroid hormone-related protein (pthrp). the gene codes for a 125-amino acid mature protein with a 35-residue prepeptide. the total gene sequence is 1.8 kb with approximately 75% noncoding. the n-terminus of the protein resembles mammalian and chicken pthrp peptides with 12 of the first 21 amino acids identical and for which there is homology with mammalian parathyroid hormone. toward the c-terminus, the nuclear transporter ...200010843788
evolution of the thyroid hormone-binding protein, transthyretin.transthyretin (ttr) belongs to a group of proteins, which includes thyroxine-binding globulin and albumin, that bind to and transport thyroid hormones in the blood. ttr is also indirectly implicated in the carriage of vitamin a through the mediation of retinol-binding protein (rbp). it was first identified in 1942 in human serum and cerebrospinal fluid and was formerly called prealbumin for its ability to migrate faster than serum albumin on electrophoresis of whole plasma. it is a single polype ...200011017772
molecular cloning and sequence of sparus aurata skeletal myosin light chains expressed in white muscle: developmental expression and thyroid regulation.two full-length cdna clones encoding the skeletal myosin light chain 2 (mlc2; 1452bp) and myosin light chain 3 (mlc3; 972bp) were isolated from a cdna library prepared from gilthead sea bream sparus aurata larvae. the mlc2 cdna encoded a predicted protein of 170 residues that was 79% identical to rabbit mlc2 over the entire length and 87% identical within the ca(2+)-binding region. the deduced amino acid sequence of mlc3 was 153 residues in length and was 91% and 69% identical to the zebrafish a ...200111551989
developmental expression, tissue distribution and hormonal regulation of fish (sparus aurata) serum retinol-binding protein.retinol-binding protein (rbp) is the specific carrier of retinol in vertebrates and forms a 1:1 complex with transthyretin (ttr). a cdna encoding serum rbp was cloned from liver and 7-day larvae of the marine fish sparus aurata. the mature protein is 176 amino acids long and shows sequence identity of 77-78%, 56%, 63% and 62% with rainbow trout, xenopus, chicken and human rbp, respectively. northern blot analysis of hepatic rbp revealed two transcripts: a major one of approximately 1.4-1.5 kb an ...200111399497
developmental expression of alcohol dehydrogenase (adh3) in zebrafish (danio rerio).alcohol dehydrogenase (adh) is the primary enzyme responsible for metabolism of ethanol to acetaldehyde. one class of adh has been described in fish, and has been found to be structurally similar to mammalian class iii adh (glutathione-dependent formaldehyde dehydrogenase) but functionally similar to class i adh (primarily responsible for ethanol metabolism). we have cloned a cdna by rt-pcr from zebrafish (danio rerio) liver representing the zebrafish adh3 gene product, with a coding region of 1 ...200111527411
characterization of a sea bream (sparus aurata) thyroid hormone receptor-beta clone expressed during embryonic and larval development.a clone encoding thyroid hormone receptor-beta (tr-beta) was isolated from a sea bream (sparus aurata) ovary cdna library. sea bream (sb)tr-beta is closely related to its counterparts from other vertebrates and, like them, preferentially binds t3 rather than t4. however, the putative sbtr-beta protein contains a nine-amino-acid insert that is also present in the corresponding proteins from flounder and salmon but absent in tr-betas from zebrafish and terrestrial vertebrates. semiquantitative rt- ...200111551120
cymothoid parasite ceratothoa parallela inflicts great losses on cultured gilthead sea bream sparus aurata in greece.for the first time ceratothoa parallela (otto, 1828), a cymothoid isopod, is reported parasitizing cage-cultured gilthead sea bream sparus aurata, in greece. the specimens observed are larvae (pullus secundus). they were found in the branchial and buccal cavity of young gilthead sea bream of 2 g mean body weight, upon introduction in the cages in an intensive cage farm facility. the species was previously known from several species of wild fish, particularly sparidae (chiefly boops boops) in the ...200111558733
characterization of pseudomonas spp. associated with spoilage of gilt-head sea bream stored under various conditions.the population dynamics of pseudomonads in gilt-head sea bream mediterranean fish (sparus aurata) stored under different conditions were studied. phenotypic analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins were performed to identify a total of 106 pseudomonas strains isolated from s. aurata stored under different temperatures (at 0, 10, and 20 degrees c) and packaging conditions (air and a modified atmosphere of 40% co(2)-30% n(2)-30% o(2)). pseudomon ...200211772610
cymothoid isopod parasites in aquaculture: a review and case study of a turkish sea bass (dicentrarchus labrax) and sea bream (sparus auratus) farm.this paper reviews the occurrence of cymothoid isopod parasitism in aquaculture, reports the first case of infection by a cymothoid isopod (ceratothoa oestroides) in turkish aquaculture, and analyses its effects on sea bass dicentrarchus labrax. analyses revealed that c. oestroides negatively affects the weights and lengths of sea bass hosts. these effects have been previously underestimated because host age has not been accounted for. the analysis of condition factors as a means of assessing pa ...200111710552
glyconjugates in epidermal, branchial and digestive mucous cells and gastric glands of gilthead sea bream, sparus aurata, senegal sole, solea senegalensis and siberian sturgeon, acipenser baeri development.epidermal, branchial and digestive mucous cells, and the gastric glands of larvae/postlarvae (from hatching until 45 days posthatching) of three fish species (two teleostean and a chondrostean) were investigated using conventional histochemical methods (periodic acid schiff -pas-, diastase-pas; alcian blue ph 0.5, 1 and 2.5) in order to distinguish neutral and acidic (carboxylated and sulphated) glycoconjugates, as well as bromophenol blue reaction for identification of proteins. additionally, t ...200111759813
oral administration of yeast, saccharomyces cerevisiae, enhances the cellular innate immune response of gilthead seabream (sparus aurata l.).the effects of including lyophilised whole yeast, saccharomyces cerevisiae, in the diet on the seabream innate immune response were investigated. gilthead seabream (sparus aurata l.) specimens were fed four different diets for 4 weeks: a commercial diet as control and the same diet supplemented with 1, 5 or 10 g/kg yeast. after 1, 2 and 4 weeks, serum complement titres, as a humoral parameter, and phagocytic, respiratory burst, myeloperoxidase and natural cytotoxic activities of head-kidney leuc ...200211867166
the gnrh system in the european sea bass (dicentrarchus labrax).the cdna sequences encoding three gnrh forms, sea bream gnrh (sbgnrh), salmon gnrh (sgnrh) and chicken gnrh ii (cgnrh ii), were cloned from the brain of european sea bass, dicentrarchus labrax. comparison of their deduced amino acid sequences to the same forms in the gilthead sea bream, sparus aurata, and striped bass, morone saxatilis, revealed high homology of the prepro-cgnrh ii (94% and 98% respectively), and prepro-sgnrh (92% to both species). the sbgnrh exhibited dissimilar identities, wit ...200211786378
hydrophobicity and adhesion to fish cells and mucus of vibrio strains isolated from infected fish.the hydrophobicity of 44 vibrio strains isolated from cultured, diseased gilt-head sea bream (sparus aurata) was determined. three different methods were used: (1) microbial adhesion to hydrocarbons (math), either with phosphate buffer or with phosphate urea magnesium sulfate (pum) buffer, (2) aggregation in the presence of salt solutions (sat), and (3) adhesion to nitrocellulose filters (ncf). the results show that experimental conditions exerted a significant influence on hydrophobicity. thus, ...200111770816
sea bream sparus aurata, an asymptomatic contagious fish host for nodavirus.during an epidemiological survey of viral encephalopathy and retinopathy (ver) in diseased sea bass dicentrarchus labrax, a nodavirus isolate was recovered from net pen-reared sea bream sparus aurata harboured in the same farming premises. after the virus was isolated and identified by immunofluorescence on ssn-1 cells, sequence analysis with a pcr product from the t4 region of the capsid protein gene indicated that the virus shared 100% identity with a pathogenic virus strain isolated from sea ...200111797913
immunocytochemical demonstration of melanotropic and adrenocorticotropic cells from the gilthead sea bream (sparus aurata l., teleostei) by light and electron microscopy: an ontogenic study.in the pituitary of gilthead sea bream, sparus aurata, melanotropic (msh) and adrenocorticotropic (acth) cells were identified at the light and electron microscopic levels using rabbit anti-synthetic alphamsh (msh) and anti-human acth (1-24) (acth) sera. the distribution of these cell types was followed from hatching to 48 months. the techniques used included the peroxidase anti-peroxidase (pap) method, conventional electron microscopy, and an immunogold technique. using pap, msh (immunoreactive ...200211884085
parathyroid hormone-related protein in lower vertebrates.the genes for parathyroid hormone-related protein (pthrp) have been cloned in two teleost fishes, cdna of sea bream (sparus aurata) and genomic dna of puffer fish (fugu rubripes). the gene sequences show that there is significant conservation of amino acid identity, with specific domains most highly conserved. the n-terminus, responsible for bone matrix lysis in mammals and chickens, is present in the fish genes with 52% sequence identity to higher vertebrate pthrp peptides; the nuclear transpor ...200211997212
photobacterium damselae ssp. piscicida: detection by direct amplification of 16s rrna gene sequences and genotypic variation as determined by amplified fragment length polymorphism (aflp).a pcr protocol for the rapid diagnosis of fish 'pasteurellosis' based on 16s rrna gene sequences was developed. the procedure combines low annealing temperature that detects low titers of photobacterium damselae but also related species, and high annealing temperature for the specific identification of p. damselae directly from infected fish. the pcr protocol was validated on 19 piscine isolates of p. damselae ssp. piscicida from different geographic regions (japan, italy, spain, greece and isra ...200212033705
molecular cloning and expression analysis of tumor necrosis factor alpha from a marine fish reveal its constitutive expression and ubiquitous nature.the tumor necrosis factor alpha ( tnf alpha) gene from the marine fish, gilthead seabream (sparus aurata l.), has been isolated by rt-pcr using degenerate primers designed against vertebrate tnf alpha conserved motifs and subsequent rapid amplification of cdna ends (race). the tnf alpha cdna consists of a 142 bp 5' untranslated region (5'utr), a single open reading frame of 762 bp, which could code for a 253 amino acid protein, and a 476-bp 3'utr. the protein sequence deduced from seabream tnf a ...200212073149
cryptosporidium molnari n. sp. (apicomplexa: cryptosporidiidae) infecting two marine fish species, sparus aurata l. and dicentrarchus labrax l.cryptosporidium molnari n. sp. is described from two teleost fish, the gilthead sea bream (sparus aurata l.) and the european sea bass (dicentrarchus labrax l.). the parasite was found mainly in the stomach epithelium and seldom in the intestine. oocysts were almost spherical, with four naked sporozoites and a prominent residuum, and measured 3.23-5.45 x 3.02-5.04 (mean 4.72 x 4.47) microm in the type host, gilthead sea bream (shape index 1-1.17, mean 1.05). sporulation was endogenous, as fully ...200212076630
olfactory sensitivity to changes in environmental [ca(2+)] in the freshwater teleost carassius auratus: an olfactory role for the ca(2+)-sensing receptor?olfactory sensitivity to changes in environmental ca(2+) has been demonstrated in two teleost species; a salmonid (oncorhynchus nerka) and a marine/estuarine perciform (sparus aurata). to assess whether this phenomenon is restricted to species that normally experience large fluctuations in external ion concentrations (e.g. moving from sea water to fresh water) or is present in a much wider range of species, we investigated olfactory ca(2+) sensitivity in the goldfish (carassius auratus), which i ...200212177141
amplified fragment length polymorphism (aflp) and biochemical typing of photobacterium damselae subsp. damselae.the aim of the present study was to characterize subspecifically photobacterium damselae subsp. damselae strains isolated from cultured sparus aurata and dicentrarchus labrax by means of phenotypic and molecular typing techniques (amplified fragment length polymorphism, aflp).200212234352
formation of short-lived multinucleated giant cells (mgcs) from cultured gilthead seabream macrophages.monocytes/macrophages obtained from the head kidney and peritoneal cavity of gilthead seabream (sparus aurata) were cultured using plates from three different manufacturers, and were maintained under different conditions. the effects on the morphology and fusion of monocytes/macrophages of initial cell loading, removal of non-adherent cells at different times after plating, and addition of serum and antibiotics were evaluated by light microscopy, and transmission (tem) and scanning (sem) electro ...200212115269
physiological changes in the spawning gilthead seabream, sparus aurata, succeeding the removal of males.the physiological effects triggered in females by the removal of males from a group of spawning fish were examined in the multiple batch spawner, the gilthead seabream, sparus aurata. one week after the removal of males, a large portion of the oocytes underwent atresia, and sporadic release of low quality eggs continued at low frequency over a period of seven weeks. the transcript levels of the three native gonadotropin releasing hormone (gnrh) forms, salmon (s)gnrh, seabream (sb)gnrh, and chick ...200212115938
osmoregulatory action of prl, gh, and cortisol in the gilthead seabream (sparus aurata l).the osmoregulatory actions of ovine prolactin (oprl), ovine growth hormone (ogh), and cortisol were tested in the euryhaline gilthead seabream sparus aurata. acclimated to sea water (sw, 40 ppt salinity, 1000 mosm/kg h(2)o) or brackish water (bw, 5 ppt, salinity, 130 mosm/kg h(2)o), injected every other day for one week (number of injections, 4) with saline (0.9% nacl), oprl (4 microg/g body weight), ogh (4 microg/g body weight) or cortisol (5 microg/g body weight), and transferred from sw to bw ...200212441119
pcr detection and pfge dna macrorestriction analyses of clinical isolates of pseudomonas anguilliseptica from winter disease outbreaks in sea bream sparus aurata.a pcr-based detection system for pseudomonas anguilliseptica was evaluated. the primer combination paf-par (forward primer paf = 5'-gacctcgccatta-3', reverse primer par = 5'-ctcagcagttttgaaag-3') gave a unique and specific amplification product of 439 bp at an annealing temperature of 46 degrees c with all the p. anguilliseptica isolates and strains (n = 56) but no amplification products were observed with any other pseudomonas species or phylogenetically related bacteria tested. the pcr assay h ...200212152901
cloning of the cdna for the putative calcium-sensing receptor and its tissue distribution in sea bream (sparus aurata).the cdna for the calcium-sensing receptor (casr) gene has been cloned from the marine teleost sparus aurata, the sea bream. the isolated clones were 3.3 kb long with an open reading frame of 2820 bp, a 5' utr of 240 bp, and 3' utr of 248 bp. the gene codes for a mature peptide of 940 amino acids which has three principal domains; the extracellular region is more than half the total protein, there is a seven-transmembrane domain, and there is a short intracellular domain. there is considerable se ...200212383439
identification of a fasl-like molecule in leucocytes of the teleost fish gilthead seabream (sparus aurata l.).the possible expression of fasl in gilthead seabream leucocytes was studied by flow cytometry, immunoblotting and immunocytochemistry, using an anti-mouse fasl monoclonal antibody. the results pointed to a cytosolic fasl-like, but not a membrane-like form, in resting leucocytes from head-kidney, thymus, spleen, blood and peritoneal exudate. immunoblotting revealed a 19kda band in resting leucocytes, while activated leucocytes showed the same band and another of 39kda. the fasl-like molecule is i ...200312477498
ontogeny, tissue distribution, and hormonal regulation of insulin-like growth factor binding protein-2 (igfbp-2) in a marine fish, sparus aurata.in this study, we have cloned insulin-like growth factor binding protein (igfbp)-2 from a marine hermaphroditic fish species, the gilthead sea bream (sparus aurata), and determined its structure, ontogeny, tissue distribution, and hormonal regulation. the sea bream igfbp-2 precursor consists of 286 amino acids, including a putative signal peptide of 22 residues and a mature protein of 264 residues. the overall sequence of sea bream preigfbp-2 is 52-39% identical to that of zebrafish, chick, mous ...200212392684
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