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1.
ABSTRACT:   Transgenic technology has been widely applied to a variety of freshwater fish species. However there are few reports on the use of this technology in commercially important marine species. In this study, the construction of expression vectors containing the β-actin promoter region for use in the red sea bream Pagrus major , a species of considerable importance to the aquaculture industry in Japan is reported. The β-actin gene was cloned from a red sea bream genomic DNA library. Recombinant plasmids were constructed by linking the 5' flanking region of the β-actin gene to the green fluorescent protein reporter gene, followed by the poly A signal sequence of simian virus 40 or the 3' flanking region the β-actin gene. Expression of these constructs was examined following microinjection into zebrafish and red sea bream embryos, and compared to that of the expression vector pXI-GFP driven by the Xenopus elongation factor 1α. The results indicated that the construct consisting of the β-actin 5'-and 3' flanking regions was the most efficacious. In future studies, it is planned to investigate the efficient condition for integration into chromosomes of the transgene.  相似文献   

2.
The development of an inert food to replace live prey during the early stages of marine fish larvae requires research in different fields and therefore a precise work strategy. Our research on this subject has been carried out in successive steps using the gilthead seabream Sparus aurata . The first step was the design of a food particle that would be well accepted and ingested by free-swimming marine larval fish during the first developmental stages. We chose microencapsulation by polymerization of the dietary protein as the most appropriate method for making the particles; different types of microcapsules were made using a basic diet containing only the major dietary components. In the second step, our aim was to keep the larvae alive in a routine rearing system in 300-L tanks, using exclusively this kind of food, long enough to detect any changes in growth, survival, or anatomical and histological status of the larvae, in order to verify whether the technological changes were positive. The third step focused on diet formulation and searching for clues to inefficient assimilation and growth. The use of ' in vitro ' digestibility techniques allowed us to detect the inhibitory effect of some diet ingredients on larval proteases and to determine more suitable sources of protein. We now have a microcapsule able to efficiently support growth and development of S. aurata larvae , at least during the first 2 weeks of life, although the larvae still need to feed on rotifers during the first 2–4 days of exogenous feeding. This microcapsule will make it possible to make advances in determining the specific nutritional requirements of larval fish.  相似文献   

3.
ABSTRACT:   High levels of free d -alanine were found in the muscle of a marine gastropod Cellana grata that inhabited the intertidal zone, and alanine racemase activity was detected in the muscle. The authors purified alanine racemase from the muscle of C. grata to characterize its enzymological properties. The molecular mass of the enzyme was estimated to be 40.5 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis and 41.4 kDa by gel filtration, suggesting that the enzyme was monomeric in nature. Kinetic experiments, performed using the purified enzyme, revealed that the Lineweaver-Burk plot for d -alanine as a substrate resulted in a K m value of 20.4 mM, and the value for l -alanine was 43.0 mM. Of the several types of amino acids tested, alanine was found to be the specific substrate for the enzyme. In the measurement of alanine racemase activity, exogenous pyridoxal 5'-phosphate (PLP) was not required for the enzyme activity; however, aminooxyacetic acid, hydroxylamine and phenylhydrazine, which inhibit PLP-dependent enzymes, strongly inhibited the enzyme activity. These results suggest that the enzyme is PLP-dependent. This is the first report on the purification and some properties of alanine racemase in a marine gastropod.  相似文献   

4.
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5.
ABSTRACT:   Fourteen bacterial strains isolated from marine environments exhibited antagonistic action against a wide range of bacteria including Vibrio spp. A double layer agar method was used for preliminary screening to determine the relative degree of growth inhibition or bacteriolysis exhibited by the isolates. Most of the antagonistic isolates were found to be Gram-negative, motile rods and were oxidase positive, and oxidative in the oxidation and fermentation test, suggesting that they are belong to the genera Pseudomonas . The antagonistic isolates lyzed the dead cells of marine Gram-negative bacteria in both plate and liquid methods. Bacteriolytic and casein hydrolytic activities were observed in the culture supernatant of the isolates. Anion exchange column chromatography (Toyopearl DEAE-650 M) was used to purify the extracellular protease produced by an antagonistic strain A1-J25a. The active fractions of protease collected from the eluted solution also exhibited bacteriolytic activity.  相似文献   

6.
ABSTRACT:   A quantitatively major collagen was isolated from the pepsin-solubilized collagen preparation of the mantle by differential salt precipitation and phosphocellulose column chromatography, and its constituent α components (α1 and α2) were purified by phosphocellulose column chromatography. The subunits were demonstrated to be genetically distinct from each other by peptide mapping and amino acid analysis. The amino acid composition calculated from those of the α1 and α2 components in 2 : 1 ratio coincided well with that of the major collagen from the mantle. These results suggest that the major collagen in the mantle of the oyster may have a heterotrimer structure (α1)2α2.  相似文献   

7.
This study was conducted to evaluate the effect of dietary inclusion of different levels of fermented macroalga, Enteromorpha prolifera, on the growth performance, digestive enzyme activities and serum non‐specific immunity of red tilapia (Oreochromis mossambicus × Oreochromis niloticus). Fish were fed the control diet or the five diets supplemented with fermented E. prolifera for 7 weeks (diets 1–5 containing 10, 20, 30, 40 and 50 g kg?1 of fermented E. prolifera respectively). The results showed that the percentage weight gain and specific growth rate (SGR) improved significantly in fish fed diets 3, 4 and 5, and regression analysis showed SGR reached the maximum in fish fed diet containing 37 g kg?1 fermented E. prolifera, while the feed conversion ratio reduced significantly in fish fed diets 2, 3, 4 and 5, and regression analysis showed it reached its minimum in fish fed diet containing 41 g kg?1 fermented E. prolifera. The pepsin, erepsin, gastric amylase, intestinal amylase, gastric lipase and intestinal lipase activities in fish fed diets 2, 3 and 4 were higher significantly than the control, with the exception of the pepsin activity in fish fed diet 4. The serum total superoxide dismutase, lysozyme, acid phosphatase and alkaline phosphatase activities enhanced significantly in fish fed diets 3 and 4. In conclusion, fermented E. prolifera had positive effects on the growth performance, digestive enzyme activities and serum non‐specific immunity of red tilapia and the recommended dose of fermented E. prolifera is 37–41 g kg?1 in the diet of red tilapia.  相似文献   

8.
ABSTRACT:   Carbon and nitrogen isotopic ratios in tissue of the bivalve corbicula ( Corbicula japonica ) and particulate organic matter (POM) were measured along a salinity gradient in the Kushida Estuary, Japan. The bivalve exhibited a gradual isotopic enrichment from the uppermost estuarine site (δ13C = −24.8‰ and δ15N = 8.6‰) to the marine site (δ13C = −16.1‰ and δ15N = 11.8‰). Using the concentration-weighted mixing model, the bivalves' food source is estimated from the isotope values for the bivalves and POM from terrestrial plants, marine phytoplankton and benthic microalgae. The results indicated that the contributions of benthic micro algae and phytoplankton were small, while terrestrial particulate matter is significantly important for the corbicula diet, although the contribution varies among sampling sites.  相似文献   

9.
Seven marine cyanobacteria were isolated from two regions of the Gulf of Thailand and evaluated by the agar diffusion method for antibacterial activity. Inhibitory compound was purified from the crude methanol extract and its structure was elucidated based on extensive spectroscopic analysis, including IR, 1D and 2D NMR spectra as well as mass spectrometry. A novel antimicrobial compound produced by the marine cyanobacterium Leptolyngbya sp. LT19 was identified to be a 2‐hydroxyethyl‐11‐hydroxyhexadec‐9‐enoate which has so far never been reported in microorganisms. Biological assays revealed that this novel compound exhibited antibacterial activities against the Gram‐negative, persistent shrimp pathogens, Vibrio harveyi and V. parahaemolyticus with minimal inhibitory concentration of 250–1000 and 350–1000 μg mL?1 respectively.  相似文献   

10.
ABSTRACT:   Among several marine antagonistic bacteria isolated from the sea water of Kagoshima Bay, Pseudoalteromonas sp. A1-J11 was found to produce anti- Vibrio substances. The anti- Vibrio substances were extracted from the culture supernatant of the strain with chloroform and isolated using reverse-phase (Cosmosil 75C18-OPN) column chromatography followed by high-pressure liquid chromatography (Mightysil RP-18 GP Aqua). Purified substances, designated as AVS-03 a , c and d showed similar ultraviolet absorption spectra with λmax at 215, 235, 315 and 327 nm in methanol. AVS-03 d , the major anti- Vibrio substance, was thermostable up to 100°C and pH stable over a pH range higher than 4.0, and also showed strong inhibitory activities, specifically against Vibrio harveyi strains.  相似文献   

11.
ABSTRACT:   Tyrosinase inhibitor-producing fungus of Trichoderma sp. strain H1-7, which was isolated from marine sediment, was investigated by phylogenetic analysis and physiological characteristics. Phylogenetic analyses of the strain were similar to Trichoderma atroviride or T. viride . Physiological characteristics of the strain were similar to T. viride , and based on these results, it was identified as T. viride . Characterization of tyrosinase inhibitory activity of a culture supernatant of the strain was investigated. The inhibitory activity of the supernatant of the strain decreased after cultivation for more than 3 days. Furthermore, sea water was not essential for the production of the tyrosinase inhibitor (TI). When TI production of the strain was compared to T. viride , the strain showed higher activity than T. viride . From this result, it seemed that the strain had characteristic features comparable to T. viride , which was isolated from the terrestrial environment. As TI production of the strain showed higher potential than that of T. viride , it is necessary to elucidate the chemical structure of TI exemplified in the present study.  相似文献   

12.
ABSTRACT:   Feeding activity was examined at high temperatures by using a demand feeder for thermally selected and normal rainbow trout Oncorhynchus mykiss strains. When the water temperature was raised in experiment 1 from 17.5 to 25.7°C at 0.3°C/day in 21 days, the average daily food consumption rate in the thermally selected strain rose to 7.1%, which was significantly higher than that of the normal strain (4.1%, P  < 0.05). The corresponding rate was also significantly higher in the thermally selected (0.8%) than in the normal strain (0.2%, P  < 0.05) of fish in experiment 2 where water temperature was raised to 24.1°C in 0.5°C/day increments. When water temperature was raised rapidly in experiment 3 from 16.7 to 21.7°C in one day and gradually to 24.4°C in 28 days at 0.1°C/day, the average daily food consumption rates were 1.0 and 0.1% for the thermally selected and normal strains, respectively, with significant difference ( P  < 0.01). These results suggest that the thermally selected strain has acquired thermal tolerance as a result of artificial selection.  相似文献   

13.
ABSTRACT:   A C-type lectin (BRA-2) isolated from the acorn barnacle Megabalanus rosa , which was a glycoprotein having an N -linked sugar chain, was deglycosylated by N -glycopeptidase F. The structure of the released sugar chains was determined by a 2-D mapping method after derivatization with a fluorescent reagent, 2-aminopyridine, to be Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc and Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc. The structures were confirmed by matrix-assisted laser desorption ionization mass spectrometry and a comparison with authentic sugar chains by high-pressure liquid chromatography. Various properties of BRA-2 were examined before and after deglycosylation. The susceptibility of BRA-2 to protease digestion was increased by deglycosylation. However, the inhibitory activity toward calcium carbonate crystallization as well as the hemagglutinating activity of deglycosylated BRA-2 was significantly decreased. These results suggest that the sugar chains of BRA-2 are important to both its structural stability and its function.  相似文献   

14.
ABSTRACT:   Mottled amago salmon Oncorhynchus masou ishikawe with both yellow- and dark-pigmented skin occurred together with typical albino individuals in a commercial farm. Out of 12 mottled fish examined by DNA content flow-cytometry and erythrocytic nucleus size, three were diploid, eight were haploid-diploid mosaic and one was diploid-triploid mosaic. This fact indicates that the mottled coloration might link to polyploid mosaicism. Genotype of diploid and non-diploid cells at the albino locus was estimated in nine mature mottled fish by observing the frequency of wild-type and albino progeny when mating to homozygous albino ( aa ). One diploid and three haploid-diploid mosaic mottled fish were presumed to have mosaic genotype with both hemizygous ' a ' and heterozygous ' Aa ' cells ( a/Aa ), because the segregation ratio between two phenotypes was 1 : 1. Three other haploid-diploid mosaic fish were presumed to have mosaic genotype with both hemizygous ' a ' and homozygous ' AA ' cells ( a/AA ), because of exclusive occurrence of wild-type phenotype in the progeny. The diploid-triploid mosaic mottled fish was presumed to have mosaic genotypes ' aa / AAA ', ' aa/AAa ' or ' aa / Aaa ', because this fish yielded only albino progeny. One diploid mottled fish produced both two phenotypes but albino embryos appeared with much more frequency than the expectation, when assuming the genotype ' Aa '. Thus, this fish was considered to have mosaic genotype ' Aa/aa '.  相似文献   

15.
ABSTRACT:   Yessotoxin (YTX) is a shellfish toxin and its contamination in bivalves has seriously damaged shellfish industries. The biogenetic origin of YTX was identified as the dinoflagellate Protoceratium reticulatum (Claparède et Lachmann) Bütschli collected in New Zealand and Yamada Bay, Iwate in Japan. Scallops cultured in Mutsu Bay, Japan, were frequently contaminated with YTX, however, occurrence of P. reticulatum in this bay and YTX production by the local strains have not been investigated. Eight strains of P. reticulatum , isolated from the bay, were cultured in the laboratory, and analyzed by fluorometric high-pressure liquid chromatography and liquid chromatography/mass spectrometry methods for YTX production and composition. All strains tested were confirmed to produce YTX, and none of them produced known YTX analogs. Toxin amount and composition differed from strain to strain. This result is also confirmation of one of the biogenetic origins of YTX in Mutsu Bay.  相似文献   

16.
ABSTRACT:   Nucleotides and Ca2+ binding to α-actin prepared from ordinary skeletal muscle of carp Cyprinus carpio was studied. When bound Ca2+ was removed with ethylenediaminetetraacetic acid, carp α-actin denatured more rapidly than chicken α-actin. Kinetic studies of the denaturation process showed that in the absence of divalent cations, the binding constants of ATP to carp and chicken actin were 5.0 × 104/M and 1.2 × 105/M, respectively. Competitive binding of Ca2+ between actin and 8-amino-2-[(2-amino-5-methylphenoxy)methyl]-6-methoxyquinoline-N,N,N',N'-tetraacetic acid (Quin 2) showed that affinity of Ca2+ for carp actin was also lower than that for chicken actin by a factor of 1.6. These results indicated that carp actin could relatively easily denature due to the low affinities of these ligands. Enthalpy changes upon ATP binding to carp and chicken actin were −65 kJ/mol and −110 kJ/mol, respectively. Thermodynamic analyses of our results revealed that the entropy change associated with ATP binding to carp actin was significantly smaller than that to chicken actin, suggesting that structural stabilization upon ATP binding was less effective in carp actin.  相似文献   

17.
In this study, we analysed the effect of population density and food concentration on the fecundity of a Mediterranean strain of Acartia tonsa to maximize egg production. During 4‐day feeding experiments, egg hatching success and faecal pellet production were also followed. The algae Rhinomonas reticulata was supplied at different concentrations corresponding to 250, 500, 1000, 1500, 2000 and 3000 μg C L?1 day?1 at the following adult copepod density: 40, 80 and 160 ind. L?1. Our results show a positive relationship between algal concentration and egg production under all experimental conditions confirming that the quantity of food strongly limits A. tonsa fecundity. Maximum egg production (57 eggs per female) was reached at the lowest density and at the maximum food concentration. Percentage of egg hatching success was not dependent on the quantity of food used. At the same food concentration, an increase in population density from 40 to 80 ind. L?1 induced an increase in faecal pellet production per couple which did not correspond to an increase in egg production, suggesting that higher energetic costs were shifted to swimming activity. Productivity of the A. tonsa Mediterranean strain is mainly limited by the quantity of food rather than by crowding conditions.  相似文献   

18.
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19.
ABSTRACT:   We investigated the effects of specific light wavelengths from light-emitting diodes (LEDs) on the growth of the dinoflagellate Heterocapsa circularisquama , which kills bivalves, and the diatom Skeletonema costatum , which is an important food source for bivalves. Growth of H. circularisquama was obviously inhibited at 590 nm and a photon flux density less than 75 μmol quanta/m2/s. However, growth of S. costatum was not suppressed by irradiance from any LEDs tested from near-ultraviolet to near-infrared wavelengths at 75 μmol quanta/m2/s. The growth rate of H. circularisquama in an experimental treatment group with irradiance provided by both cool-white fluorescent lamps (12:12 h L : D cycle) and a 590-nm LED (continuous irradiance) was 0.43/day. In the control group with irradiance provided only by cool-white fluorescent lamps (12:12 h L : D cycle), the growth rate was 0.63/day, indicating that growth of H. circularisquama was suppressed by 590 nm (less than 75 μmol quanta/m2/s) irradiance from the LED and the continuous irradiance. The use of 590-nm LEDs in bivalve culture at irradiance levels less than 75 μmol quanta/m2/s might encourage the growth of the useful diatom S. costatum without stimulating growth of the harmful dinoflagellate H. circularisquama .  相似文献   

20.
ABSTRACT:   Seasonal changes in the infection state of marine birnavirus (MABV) in Japanese flounder Paralichthys olivaceus and in rearing sea water are described. Sea water and 10–11 healthy fish were sampled monthly from April 2002 to February 2003. The MABV genome was detected throughout the year in > 80% of the fish examined at each sampling. The virus was isolated from the liver, kidney, and spleen, but not from the brain. The detection rate in each organ increased from April to October, and then decreased. Detection of virus antigen by the indirect fluorescent antibody technique also showed that the virus was present from spring to autumn (June–September) in the liver, kidney, and spleen, but not the brain. Sequence analysis of the MABV genome at the VP2–NS region revealed two specific mutations compared to the standard yellowtail strain (Y-6). It is suggested that the infection state of MABV in Japanese flounder changes to a latent or persistent infection after autumn. MABV was detected in sea water between September and February, suggesting that virus particles in the environment are relatively higher during cool seasons.  相似文献   

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