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[Fish nutrition. Brief review for the practitioner regarding food and ornamental fish].

In behalf of the practitioner a short survey is given about nutrition of fish and shrimps. Interrelations between fish development and influences by their water ecosystem concerning temperature, mineral and gas levels, waterflow et cetera together with the supply of food are mentioned. Further comments are given about common nutritional needs of different species in connection with growth, bodyweight et cetera and purpose of fishkeeping for instance for consumption, reproduction or ornamentally in different systems from aquarium to lagoon. More specifically feed formulation from raw materials to complete diets and nutritional analyses in respect to allowances for maintenance and growth are discussed. Data about fresh- and saltwater species from tropical to arctic circumstances mentioning trout, salmon, catfish, carp, sea bass, shrimp et cetera are given. Some remarks about manufacturing, and application of complete compound feeds are added to the survey. In behalf of further study or for enhancement of knowledge a number of books are recommended in a list of references.

Animal Feed↗

[Health effects of fish oil and fish oil supplements: consumption advice sustained].

Omega-3 fatty acids are thought to have beneficial effects on atherosclerosis. Recently, the results ofa systematic review on the health effects of omega-3 fatty acids were published. The risk of cardiovascular death in subjects randomised to taking omega-3 fats was not significantly decreased: relative risk = 0.87 (95% CI: 0.73-1.03). In a 2004 Cochrane review, the same authors concluded that there was a significant beneficial effect on total mortality, combined cardiovascular endpoints and cancer. Inclusion of the recent 'Diet and reinfarction trial' (DART-2-trial) had a decisive negative effect on the results of the recent meta-analysis. Excluding this study resulted in a relative risk of cardiovascular death of 0.83 (95% CI: 0.75-0.91). Since the DART-2 study had methodological shortcomings, it still appears that recommending sufficient intake of omega-3 fatty acids is justifiable, preferably by eating fish but ifnecessary by using fish oil supplements, especially for patients at high risk of cardiovascular disease.

Cardiovascular Diseases↗

[Biogenic amine contents in fish and fish preserves].

A method developed on the basis of highly efficient liquid chromatography of dansyl derivatives was used to study the content of biogenic amines (tyramine, histamine, cadaverine, putrescine, spermine and spermidine) in fish and canned fish. It was found that only 7 out of 149 samples contained histamine levels exceeding maximum permissible concentration (salted herring, smoked sardines, and salted Siberian salmon). No correlation was recorded between the increase of spermine content and spermidine.

Amphetamine↗

[Detection of the hepatitis A virus in fish at a fish farm].

In this study, we have researched enteroviruses in digestive tract and gills of 106 fishes from farming station, belonging to the five most eaten species in Ivory Coast (Chrisychthys walkeri, Chrisychthys nigrodigitatus, Tilapia nilotica, Tilapia guineensis, Sarotherodon melanotheron). Hepatitis A virus was found in 12 fishes (11.3%), five times in gills, three times in digestive tract and four times in both. The two species apparently the more contaminated, were Chrisychthys nigrodigitatus, Sarotherodon melanotheron with a 4.7% contamination rate for the both.

Animals↗

Comparative antioxidant enzyme study in freshwater fishes. II. Distribution of antioxidant enzymes and lipid peroxidation in omnivorous fish organs.

Comparative studies were performed on the activities of superoxide dismutases, catalase and glutathione peroxidase in organ homogenates from three omnivorous fishes, the barbel, crucian carp and common carp. The lipid peroxidation and protein contents of organ homogenates were also compared. These comparative measurements primarily provide control values for subsequent toxicological examinations. The highest total superoxide dismutase activities were found in the liver or roe, kidney, heart and spleen in every cases. The antioxidant enzyme activities and other studied parameters of the organ homogenates partly appear to depend on the feeding mode, but are rather characteristic of the fish variety.

Animals↗

Studies on the occurrence of actinosporean stages of fish myxosporeans in a fish farm of Hungary, with the description of triactinomyxon, raabeia, aurantiactinomyxon and neoactinomyxon types.

Actinosporean infection of the oligochaete fauna living in the mud and on the vegetation of fish ponds used for rearing common carp in polyculture was studied during a one-year survey at a fish farm in Hungary, located south of Budapest. Twenty-eight actinospore types were isolated from the oligochaetes Tubifex tubifex, Branchiura a sowerbyi, Limnodrilus hoffmeisteri, Nais elinguis, and Stylaria lacustris collected during the survey, which could be classified into the triactinomyxon, raabeia, aurantiactinomyxon and neoactinomyxon groups. Drawings depicting individual actinospore types are presented on plates and their characteristic dimensions have been summarised in tables. The prevalence and seasonality of actinosporean infections observed in the various oligochaete species have been illustrated graphically. Infection by actinospores showed a pronounced seasonality. In the spring, summer and autumn the prevalence of raabeia infection in Branchiura exceeded 90%, while in the winter it dropped to 42%. A similar phenomenon could be observed for aurantiactinomyxon infection, while neoactinomyxon infection reached its peak in the autumn. In Tubifex, Limnodrilus, Nais and Stylaria species the peak of actinosporean infection occurred, with minor differences, in the spring and summer. Actinosporean infection in the individual Oligochaeta species showed much higher prevalence values than had been reported in the literature, which can be explained by the novelty of the examination technique used. It cannot be decided with absolute certainty which myxosporean developmental stage the different actinospore types described during this survey represent of the species of Myxosporea parasitic in the given ponds. This would require an experimental study for which the data presented here may serve as a basis.

Animals↗

Rapid detection of 17p11.2 rearrangements by FISH without cell culture (direct FISH, DFISH): a prospective study of 130 patients with inherited peripheral neuropathies.

Charcot-Marie-Tooth (CMT) disease and hereditary neuropathy with pressure palsies (HNPP) are two frequent hereditary motor and sensory neuropathies. CMT is characterized by slowly progressive weakness and atrophy, primarily in peroneal and distal leg muscles. The most frequent form, CMT1A, is due, in most cases, to the duplication of a 1.5 Mb region on chromosome 17p11.2 containing the peripheral myelin protein 22 gene (PMP22). The phenotype seems to result from dosage of the PMP22 gene. This hypothesis is reinforced by the existence of HNPP, which is clinically characterized by various recurrent truncular palsies or sensory loss precipitated by minor trauma, which is caused by deletion of the same 1.5 Mb region in 17p11.2. In clinical practice, the detection of the duplication or the deletion in 17p11.2, which permits a positive diagnosis, is still performed by time consuming methods (Southern blot or various combinations of molecular tools). We developed a method for the rapid detection of 17p11.2 rearrangements, using "direct FISH" and PRINS analyses, which does not require cell culture. In a prospective study of 92 patients with CMT and 38 with suspected HNPP, we compared this new technique to classical strategies like Southern blot. The results demonstrate the high sensitivity and specificity of the new FISH technique for the diagnosis of CMT1A and HNPP. Moreover, because of its simplicity and rapidity, this technique provides a useful alternative to the molecular approaches that have been used to diagnose segmental aneusomies, especially in the case of duplications that often go undetected.

Charcot-Marie-Tooth Disease↗

Metaphase FISHing of transgenic mice recommended: FISH and SKY define BAC-mediated balanced translocation.

The evolving trend to use larger transgenes and their associated increased chance of unexpected genetic events mandates more careful characterization of transgenic mice. In characterizing our five new mouse strains transgenic for the BAC, bEMS4, we have identified the highest copy number reported to date: the stable incorporation of approximately 40 copies of a 194-kb expressed transgene in a single insertion site. We caution, however, that standard molecular techniques failed to identify a balanced translocation in another strain, and an inappropriate site of insertion in a third. Molecular cytogenetic analysis using metaphase FISH was the minimum level of characterization needed to reveal these unexpected genetic events. In addition, we combined FISH and SKY to identify the transgene at the breakpoints of the balanced translocation, t(3;9). This is the first description of a BAC-mediated chromosomal rearrangement and the first application of SKY to identify transgene-induced chromosomal rearrangements.

Animals↗

A novel 90-kDa stress protein induced in fish cells by fish rhabdovirus infection.

A 90 kDa cellular protein in a fish cell, CHSE-214, showed increased expression by the infection of infectious hematopoietic necrosis virus (IHNV), heat shock, 2-mercaptoethanol, copper sulfate, and cadmium sulfate, and was detected in various kinds of cells such as human, rat, and mouse cells. The molecular mass of the 90 kDa protein was different from those of the hsp90 and grp94. In addition, all the anti-stress protein MAbs did not react with the 90 kDa protein. Finally, the subcellular distribution of the 90 kDa protein, determined by Western blots of subcellular fractions, was found to be mainly nuclear, both in normal and IHNV-infected CHSE-214 cells. The present results indicate that the 90 kDa protein is a kind of stress protein. However, based on its molecular mass, antigenic characteristics, and subcellular distribution, it is likely that this protein is a novel stress protein that has not been previously described in animal systems, especially in fish systems.

Animals↗

Adhesion and ingestion activities of fish phagocytes induced by bacterium Aeromonas salmonicida can be distinguished and directly measured from highly diluted whole blood of fish.

The phagocytes of fish play an important role in innate host defense against bacterial infection, and participate in various immunoregulatory processes. Here, we investigated the effects of various opsonins in the ingestion and adhesion processes by examining respiratory burst (RB) activity in blood and head kidney (HK) fish phagocytes. RB activity was induced in rainbow trout phagocytes with the bacterium Aeromonas salmonicida (strain MT004) in the presence of various opsonins [purified antibodies (Ab), immune serum (IS), normal serum (NS) and heat-inactivated immune serum (HI-IS)], and measured in terms of luminol-amplified chemiluminescence (CL) emission at 20 degrees C for 210 min. The RB activity of blood phagocytes was measured directly from highly diluted whole blood and compared to that observed in isolated head kidney (HK) phagocytes measured under similar conditions. In addition, the extracellular RB activity of adhesion (extracellular degranulation) and the intracellular RB activity of ingestion were distinguished through their inhibition by gelatin and cytochalasin D. Our results showed that the first CL peak appeared within 50 min, and decreased or vanished when gelatin was added to the reaction or when the active complement was destroyed by heating. The second CL peak appeared after 50 min, depending on the utilized opsonin, and vanished when cytochalasin D was added to the reaction. Our results indicate that adhesion and ingestion compete for consumption of reactive oxygen intermediates. Specific IgM without an active complement was a relatively inefficient opsonin, whereas specific IgM with an active complement increased the magnitude of ingestion-mediated RB activity and accelerated the ingestion of target bacteria. Taken together, these results indicate that adhesion and ingestion responses competed for limited phagocyte resources and that the bacterial uptake by blood phagocytes can be measured directly from highly diluted blood.

Aeromonas salmonicida↗

Characterization of a pituitary GnRH-receptor from a perciform fish, Morone saxatilis: functional expression in a fish cell line.

Gonadotropin-releasing hormones (GnRHs) bind to the specific receptor present on the gonadotrophs to activate the synthesis and release of gonadotropins (follicle stimulating hormone or FSH and luteinizing hormone or LH), which in turn control gonadal maturation, gametogenesis and gamete release. Perciform species have three endogenous GnRHs. The main objective of this study was to characterize the gonadotropin-releasing hormone receptor (GnRH-R) present in the pituitary of a perciform species, striped bass (Morone saxatilis) and demonstrate how it interacts with its potential ligand. In this study, a cDNA for GnRH-R from the pituitaries of striped bass was cloned. The cloned cDNA has an open reading frame (ORF) that codes for a 419 amino acids peptide. Like other G-protein coupled receptors including the non-mammalian GnRH-Rs, the peptide has seven putative transmembrane domains and a C-terminal tail. Comparative analysis of the amino acid sequence of striped bass (stb) GnRH-R shows 38-87% similarity with the known GnRH-Rs. A Northern blot analysis revealed a single GnRH-R transcript in the pituitary; however, its expression in various extrapituitary tissues was demonstrated by a reverse-transcription-PCR (RT-PCR). Functionally, upon induction by endogenous forms of GnRHs (seabream, chicken II and salmon GnRHs) and a mammalian GnRH-agonist, the recombinant stbGnRH-R mediated a reporter gene (luciferase) activity in a fish cell line (CHSE-214). A real-time relative quantitation method established that significantly higher (P<0.05) levels of stbGnRH-R mRNA were present in the pituitaries of striped bass with advanced stages of ovarian development, compared to the pituitaries of fish with less developed ovaries.

Amino Acid Motifs↗

Estimation of conditional stability constant for copper binding to fish gill surface with consideration of chemistry of the fish gill microenvironment.

Binding-site concentration and conditional stability constants for copper and fish gill surface interactions were calculated based on the data from the literature. Six scenarios were modeled by including or excluding pH and alkalinity differences between the fish gill microenvironment and the bulk solution and the presence of free mucus in the calculation. We demonstrate that changes in pH or alkalinity, or both, for model input had only a slight influence on the calculated results because of the small difference in pH and alkalinity between the gill microenvironment and the bulk solution under the specific experimental conditions. Inclusion of free mucus in the model, however, led to a large change in the final results. For example, with consideration of free mucus and changes in pH and alkalinity in the model, the calculated site concentration and the stability constant were 0.022 micromol/g wet tissue and log K=8.77, respectively, compared to 0.026 micromol/g wet tissue with log K=7.78 without free mucus and pH/alkalinity change.

Animals↗

Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH).

When surveying the karyotype diversity of European loaches of the genus Cobitis to identify species involved in hybrid polyploid complexes, an extensive polymorphism in number and location of NORs was discovered in C. vardarensis using Ag-staining, C-banding, CMA3-fluorescence and fluorescence in situ hybridization (FISH). This species had 2n = 50, the karyotype contained 13 pairs of metacentric, 10 pairs of submetacentric and two pairs of subtelocentric chromosomes. The NOR-bearing chromosomes included one medium-sized metacentric pair with a large CMA3-positive heterochromatic pericentromeric block, one small metacentric as well as one large submetacentric pairs. Ribosomal sites were always located in telomeres of these chromosomes. Each of the pair of NOR-bearing chromosomes occurred in three variants - (1) presence and/or (2) absence of NORs on both homologues and (3) heterozygous combination where only one of the homologues bears NORs. Altogether, 10 different NOR cytotypes from 27 theoretically possible ones were discovered among 20 indviduals examined. The number of NORs ranged from two to five per specimen. The results regarding the number and locations of NORs as revealed by banding techniques were confirmed using FISH with rDNA probe. NOR sites were of CMA3-positive, suggesting that ribosomal sites are associated with GC-rich DNA. Very similar structural polymorphism with multiple NORs is expressed in the Danubian loach C. elongatoides indicating a close relationship between both species.

Animals↗

Karyotyping mouse chromosomes by multiplex-FISH (M-FISH).

Karyotyping of mouse chromosomes is a skillful art, which is laborious work even for experienced cytogeneticists. With the growing number of mouse models for human diseases, there is an increasing demand for automated mouse karyotyping systems. Here, such a karyotyping system for mouse chromosomes based on the multiplex-fluorescence in-situ hybridization (M-FISH) technology is shown. The system was tested on a number of individual mice with numerical and structural aberrations and its reproducibility and robustness verified. Mouse M-FISH should be a valuable tool for the analysis of chromosomal rearrangements in mice.

Animals↗

Cryptic translocation identification in human and mouse using several telomeric multiplex fish (TM-FISH) strategies.

Experimental data published in recent years showed that up to 10% of all cases of mild to severe idiopathic mental retardation may result from small rearrangements of the subtelomeric regions of human chromosomes. To detect such cryptic translocations, we developed a "telomeric" multiplex fluorescence in situ hybridization (M-FISH) assay, using a set of previously published and commercially available subtelomeric probes. This set of probes includes 41 cosmid/PAC/P1 clones located from less than 100 kilobases to approximately 1 megabase from the end of the chromosomes. Similarly, a published mouse probe set, comprised of BACs hybridizing to the closest known marker toward the centromere and telomere of each mouse chromosome, was used to develop a mouse-specific "telomeric" M-FISH. Three different combinatorial labeling strategies were used to simultaneously detect all human subtelomeric regions on one slide. The simplest approach uses only three fluors and can be performed in laboratories lacking sophisticated imaging equipment or personnel highly trained in cytogenetics. A standard fluorescence microscope equipped with only three filters is sufficient. Fluor-dUTPs and labeled probes can be custom made, thus dramatically reducing costs. Images can be prepared using imaging software (Adobe Photoshop) and analysis performed by simple visual inspection.

Animals↗

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 the absence of MgEGTA their binding capacities generally increased, presumably as a result of classical-pathway activation by the natural antibodies present in the serum. The presence of EDTA abrogated the binding of all C3 isoforms to the various surfaces tested. The C3 isoforms differed in the efficiency of their binding to complement-activating surfaces: the two most abundant C3 isoforms (C3-1 and C3-2) bound to zymosan as well as to sheep and rabbit erythrocyte ghosts, whereas C3-3, C3-4 and C3-5 were unable to bind to zymosan. Upon complement activation, all five C3 isoforms were cleaved to 'iC3b' by factor H and I-like proteins, generating fragments similar to those generated from C3 molecules of other species. Furthermore the degradation of methylamine-hydrolysed C3 isoforms to iC3b was significantly inhibited by EDTA. The structural and functional diversity that we have observed in the C3 isoforms of S. aurata would increase the capacity of this fish to recognize a broader spectrum of potential pathogens and reinforce a specific immune response, which in fish is delayed compared with that of higher vertebrates, and is based on a single Ig type (IgM).

Animals↗

Chromosome assignment of six dog genes by FISH, and correlation with dog-human Zoo-FISH data.

Cross-species chromosome painting analyses have recently demonstrated the presence of regions of conserved synteny between the human and domestic dog genomes, aiding the search for candidate genes for inherited traits. Concerted efforts to subchromosomally assign substantial numbers of dog gene sequences are now needed in order to refine these comparative data, both in terms of marker density and resolution. We have developed novel PCR markers representing three dog genes (ALB, FOS, HNRPA2B1) for which no sequence or mapping data were previously available, to our knowledge. These, in addition to three gene markers previously described (ALDOA, RPE65, VCAM1), were used to isolate and chromosomally assign corresponding large insert genomic clones by fluorescence in situ hybridization (FISH). Chromosome assignments for these six dog genes are discussed in terms of those of the human orthologues, and correlated with existing comparative mapping information, identifying one apparent exception to existing Zoo-FISH data, and aiding refinement of the boundaries of conserved chromosome segments in both genomes.

Animals↗

Molecular characterization of fish neurolin: a growth-associated cell surface protein and member of the immunoglobulin superfamily in the fish retinotectal system with similarities to chick protein DM-GRASP/SC-1/BEN.

We have used the polymerase chain reaction to isolate cDNAs coding for goldfish and zebrafish neurolin, a previously identified 86 kDa cell surface glycoprotein in the goldfish visual system. Sequence analysis demonstrates that neurolin belongs to the immunoglobulin superfamily and is 51% similar to the chick cell adhesion molecule DM-GRASP, SC-1, BEN. Northern analysis with a riboprobe coding for the C-terminus of neurolin detected two mRNAs of 3.7 kb and 3.3 kb in both embryonic and adult goldfish. Several monoclonal and polyclonal antibodies were generated against immunopurified goldfish neurolin and two are shown to crossreact with zebrafish proteins. Both antibodies identify a zebrafish protein of the same molecular weight as goldfish neurolin on immunoblots. Immunohistochemical studies with these antibodies in the zebrafish retinotectal system demonstrate labeling on young ganglion cells and growing retinal axons in a pattern similar to that found in goldfish. The similarity of neurolin to a known cell adhesion molecule, its expression on developing retinal ganglion cells and axons in both embryos and adult fish, and its re-expression during retinal axon regeneration in the goldfish suggests that neurolin is important during axonal growth in the fish central nervous system.

Activated-Leukocyte Cell Adhesion Molecule↗