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[Fish silage from shrimp by catch fish: I. Preparation and biological evaluation].

Fish silage was prepared from some fish species of the shrimp by-catch caught in Golfo Triste, Carabobo, Venezuela. Fish were ground until a particle size that would guarantee the liquefaction process was achieved. The process was accelerated by the incorporation of a 3.5% w/w acid mixture, formed by diluted sulfuric acid (1:3) and concentrated formic acid, in a proportion of 1:4. The silage process occurred spontaneously at room temperature during 17 days. Chemical, physical and microbiological tests were conducted in order to characterize both the raw material and the final product. The raw material used had adequate levels of freshness, so that it was suitable for use, thus producing a first-grade silage. A first biological evaluation was carried out in rats in order to estimate the protein quality of the silage by means of certain biological parameters such as PER, NPR and apparent digestibility percentage of the protein. The results of the first experimental period demonstrated that the fish silage protein was of optimal quality, since the biological response in rats was similar to that of the control group fed the casein diet.

Animal Nutritional Physiological Phenomena↗

[Use of fish cell cultures for toxicity determination in order to reduce and replace the fish tests].

Dispersion of liver tissue from rainbow trout at 10 and 25 degrees C resultet in cells which were cultivated in vitro at 20 degrees C regularly. Using this method a cell line (R1) has been established useful for testing the toxicity of chemical substances and waste water. The toxicity of samples influenced the morphology, growth and the living duration of the cells. These parameters gave a clear answer about the toxicity of samples testet in 1-12 h. The tests on the vitality of cells (using trypan blue) and the release of LDH did not show an agreement with the cytotoxicity in all cases. From 106 samples of waste water were 53.8% cytotoxic but only 35% showed a toxicity for fish. 93.44% of 61 toxic samples of waste water showed cytotoxicity, only 60.65% of the samples were fish-toxic. According to the results obtained it is concluded that fish cell tissue cultures are useful tools for determination of the toxicity of chemical substances and waste water. Compared with the fish test the cytotoxicity test ist more sensitive and it reduces the material, time and money needed.

Animals↗

Studies on the bacterial flora of fish which are potential pathogens for human. Isolation of various potential human pathogenic organisms from different parts of fish and their significance in initiating human diseases.

Freshwater fish of different categories, collected from market of Dhaka City of Bangladesh, were studied for the bacterial flora. Potential human pathogens were isolated from about eighty-five percent of the fish studied. Organisms isolated were Aeromonas spp., (69.2%), Vibrio spp., (42.3%), Plesiomonas shigelloides (35.9%) and Escherichia coli (16.7%). The high association of potential human pathogens among the freshwater fish suggest that if fish are handled improperly or if consumed undercooked or uncooked may cause various diseases to susceptible individual.

Animals↗

Liquid chromatographic determination of oxytetracycline in edible fish fillets from six species of fish.

The approved use of oxytetracycline (OTC) in U.S. aquaculture is limited to specific diseases in salmonids and channel catfish. OTC may also be effective in controlling diseases in other fish species important to public aquaculture, but before approved use of OTC can be augmented, an analytical method for determining OTC in fillet tissue from multiple species of fish will be required to support residue depletion studies. The objective of this study was to develop and validate a liquid chromatographic (LC) method that is accurate, precise, and sensitive for OTC in edible fillets from multiple species of fish. Homogenized fillet tissues from walleye, Atlantic salmon, striped bass, white sturgeon, rainbow trout, and channel catfish were fortified with OTC at nominal concentrations of 10, 20, 100, 1000, and 5000 ng/g. In tissues fortified with OTC at 100, 1000, and 5000 ng/g, mean recoveries ranged from 83 to 90%, and relative standard deviations (RSDs) ranged from 0.9 to 5.8%. In all other tissues, mean recoveries ranged from 59 to 98%, and RSDs ranged from 3.3 to 20%. Method quantitation limits ranged from 6 to 22 ng/g for the 6 species. The LC parameters produced easily integratable OTC peaks without coelution of endogenous compounds. The method is accurate, precise, and sensitive for OTC in fillet tissue from 6 species of fish from 5 phylogenetically diverse groups.

Animals↗

Detection of the fish pathogen Flavobacterium psychrophilum in water from fish farms.

Rainbow trout fry syndrome and cold-water disease are serious diseases in farmed salmonid fish. In the present study, three methods were compared, for the detection of the causative pathogen, Flavobacterium psychrophilum in water. The methods included traditional agar plate cultivation on tryptone yeast extract salts (TYES) agar, immunofluorescence antibody technique (IFAT) and nested PCR. The three methods were subsequently used for the detection of F. psychrophilum from fish farm environments. The nested PCR was the most sensitive method used for a detection of F. psychrophilum. As low as 3 CFU estimated by agar plate cultivation or 41 cells estimated by IFAT of F. psychrophilum per ml of non-sterile well water were needed for a detection using the nested PCR method. The obtained detection limits for the agar plate cultivation and the IFAT was 32 CFU/ml and 410 cells/ml, respectively. Using IFAT and nested PCR F. psychrophilum was detected most frequently in water samples from fish farms, but the pathogen was isolated from only a few samples using agar plate cultivation. In the present study IFAT and nested PCR proved to be rapid, specific and sensitive methods compared to traditional agar plate cultivation for the detection of F. psychrophilum from environmental samples. It is suggested that IFAT and nested PCR provide effective tools for the examination of F. psychrophilum in the environment.

Agar↗

Estimation of microcystins in the freshwater fish Oreochromis niloticus in an Egyptian fish farm containing a Microcystis bloom.

Microcystins (MCYSTs) that accumulated in different organs of the freshwater fish Oreochromis niloticus, collected from a fish farm in Egypt containing heavy blooms of Microcystis aeruginosa, were investigated using an enzyme-linked immunosorbent assay (ELISA). The distribution of MCYSTs in the organs varied significantly. The highest MCYST level was recorded in the guts (821 ng/g fresh weight), followed by the livers (531.8 ng/g) and kidneys (400 ng/g). Smaller amounts of MCYST were detected in muscles (102 ng/g). The present study suggests that fish farms should be monitored for the presence of toxic cyanobacterial blooms to minimize the exposure of fish to potent hepatotoxins.

Animals↗

Using a new inbred fish model and cultured fish tissue cells to study Aeromonas hydrophila and Yersinia ruckeri pathogenesis.

An inbred strain of the southern platyfish, Xiphophorus maculatus, was used as a host for Aeromonas hydrophila and Yersinia ruckeri infections. The infections were initiated by holding the platyfish in inoculation baths containing dilutions of virulent A. hydrophila or Y. ruckeri strains. Inoculating the platyfish in this manner resulted in a dose-dependent mortality over a range of bacterial input from 10(5) to 10(8) A. hydrophila and 10(6) to 10(8) Y. ruckeri/ml. Clinical manifestations of A. hydrophila infections were noted in infected platyfish that eventually died, but not in platyfish that survived. In this model, the Y. ruckeri infected fish died before obvious signs of infection were detected. The A. hydrophila strain used to establish the infections was recovered from the kidney and intestine of infected fish that died, but not from survivors receiving the same inoculation dose. Both infective bacteria were tested for the ability to invade a number of different fish and human cultured cells. A hydrophila strain TF7 did not invade of the cells tested, whereas the Y. ruckeri strain invaded fish derived cultured cells, but not human derived Hep-2 cells.

Aeromonas hydrophila↗

Detection of ALK gene rearrangements in formalin-fixed, paraffin-embedded tissue using a fluorescence in situ hybridization (FISH) probe: a search for optimum conditions of tissue archiving and preparation for FISH.

BACKGROUND: It is widely known that the efficiency of fluorescence in situ hybridization (FISH) probes applied to formalin-fixed, paraffin-embedded tissues is affected by the conditions under which the tissues are fixed and embedded. However, relatively few studies address exactly how tissue archiving conditions affect the performance of FISH probes. We report our experience based on use of an ALK FISH probe, during the validation of its diagnostic utility. METHODS: We applied the probe to 77 formalin-fixed, paraffin-embedded tissue blocks archived from 1991 through to 2000, and studied the interrelationship between the archival age (which ranged up to 10 years), type and condition of tissue, duration required for optimum hydrolysis, and obtainability of hybridization signals. RESULTS: We found that as archival age and tissue collagen content increased, not only did hydrolysis times have to be prolonged in order to yield interpretable hybridization signals, but also the likelihood of blocks becoming non-signaling increased. The most striking positive correlations were seen between the archival age of signaling lymphoid blocks and their requisite hydrolysis times. CONCLUSIONS: The difficulty in applying FISH on archival tissue increases with its archival age and collagen content, and may necessitate changes in laboratory protocol accordingly.

Anaplastic Lymphoma Kinase↗

Biochemical responses of fish sac fry and a primary cell culture of fish hepatocytes exposed to polychlorinated naphthalenes.

Chlorinated naphthalenes are planar halogenated aromatic compounds, which are widespread in the environment. Knowledge of their biochemical and toxicological actions in aquatic biota is, however, limited. The objective of this study was to assess the toxicity of highly chlorinated naphthalene congeners found in the aquatic environment on fish sac fry and to study their effects on xenobiotic metabolizing enzymes (CYP) using a short-term primary culture of fish hepatocytes and liver microsomes. A few days after hatching, rainbow trout sac fry were administered either Hallovax 1014, a mixture of 1,2,3,4,6,7-hexachloronaphthalene and 1,2,3,5,6, 7-hexachloronaphthalene (HxCN-mix), or 1,2,3,4,5,6, 7-heptachloronaphthalene (HpCN) (0.08, 0.8, and 4 microg/sac fry injected into the yolk sac). The exposure was terminated 2 weeks later. The naphthalene preparations did not cause any clinical signs of toxicity or difference in mortality rates between the control and treated groups. Immunohistochemical analysis of CYP1A expression in the treated sac fry revealed that staining was most pronounced in the hepatocytes and thereafter in kidney tubular epithelial cells. Moderate CYP1A staining was also seen in the mucosal epithelium of pyloric caecae and mild staining in the epithelium of olfactory organ. Staining in control sac fry was weak or absent. Exposure of the primary cell culture of trout hepatocytes to a low doses (</=10 ng/ml) of the chlorinated naphthalenes increased significantly CYP1A-associated EROD activity and CYP1A mRNA content, HxCN-mix being the most potent and thereafter HpCN and Hallovax 1014. The higher doses (50-100 ng/ml) of each naphthalene also inhibited EROD activity. However, the content of CYP1A mRNA or the intensity of the CYP1A protein band (58 kDa) recognized by anti-trout CYP1A peptide antibodies were not decreased with increasing polychlorinated naphthalene (PCN) concentration, indicating that the inhibition was not due to reduced protein synthesis. Furthermore, in vitro analyses of the inhibitory potential of PCNs on CYP1A activity with trout liver microsomes suggested that these naphthalene preparations may be CYP1A substrates and act as competitive inhibitors of CYP1A catalyst. Our results demonstrate that highly chlorinated naphthalenes are potent modulators of fish CYP1A enzyme and suggest that hepatocytes and tubular epithelial cells are the cell types that may be vulnerable to their metabolic products for cell injury in fish sac fry.

Animals↗

Fish embryos as teratogenicity screens: a comparison of embryotoxicity between fish and birds.

Early life stage (ELS) toxicity experiments were carried out with zebra fish (Brachydanio rerio) and rainbow trout (Salmo gairdneri) and 10 chemicals used in the rubber industry. Several of these chemicals appeared to be teratogenic. A good correlation (r = 0.95) was found between the 7-day EC50 for zebra fish and the 60-day EC50 for rainbow trout for total embryotoxicity (embryolethality and malformations). The S. gairdneri test appeared to be slightly more sensitive than the test with B. rerio. It is therefore concluded that this short-term test is a good alternative for the long-term test with S. gairdneri. A remarkably good correlation (r = 0.90) was found between the ED50 for chicken embryotoxicity reported in the literature and the EC50 for embryotoxicity for both zebra fish and rainbow trout. This may, among other things, be explained by similarities in embryonic development and the absence of maternal and placental metabolism of the toxicants in tests with eggs of both fish and birds. It may therefore be concluded that both the short-term ELS test with B. rerio and the chicken egg test have the same predictive power for mammalian teratogenicity; i.e., both are suitable screening tests for direct-acting teratogens.

Animals↗

Cytogenetic studies of a series of 43 consecutive secondary myelodysplastic syndromes/acute myeloid leukemias: conventional cytogenetics, FISH, and multiplex FISH.

We report a series of 43 consecutive therapy-related myelodysplastic syndromes (t-MDS) or acute myeloid leukemias (t-AML) observed for 6 years. This series consisted of 26 women and 17 men, ages ranging from 9 to 85 years. These cases were classified into three groups according to the primary diagnosis. Conventional cytogenetic and fluorescent in situ hybridization (FISH)/ multiplex FISH (M-FISH) methods were used to analyze cytogenetic characteristics of secondary MDS/AML. The features of chromosomal abnormalities were linked to the nature of the therapy and protocols used. A considerable proportion of recurrent balanced translocations characterized t-AML secondary to therapy. FISH techniques showed that conventional cytogenetics often underestimated associated translocations; some deletions were in fact derivative chromosomes associated with deletions. After treatment for lymphomas and chronic myeloproliferative diseases, there were more complex unbalanced abnormalities than the control group. Compared to other series, recurrent translocations appeared to be more numerous (25%), probably reflecting an evolution of therapeutic modalities.

Adolescent↗

Preliminary data on polybrominated diphenyl ethers (PBDEs) in farmed fish tissues (Salmo salar) and fish feed in Southern Chile.

Polybrominated diphenyl ethers (PBDEs) have become an issue of global concern. Recent studies have shown that farmed salmon can accumulate high levels of brominated compounds in their tissues and consequently there is a growing concern on its industrial and public health impacts. Little information is found in the international literature on PBDEs in the biotic compartment of the Southern Hemisphere. This paper reports the levels of several PBDE congeners found in the tissues of farmed fish from five different farming areas of Southern Chile. PBDEs were analyzed by HRGC-MS. More analytical data were obtained by analyzing these same pollutants in fish feed. Our results indicate a general trend of PBDE levels averaging 1.46 ng g(-1) wet weight (wwt). The observed congeneric distribution that resulted was quite similar to data previously reported in the open literature. PBDE profiles were found to be dominated by BDE 47. No correlation was observed between levels found in the tissues and the lipid content in such tissues, although a high correlation with the fish feed data was observed indicating that this could probably be the main PDBE entry source into fish, although other sources cannot be excluded. Even though the samples were obtained from different geographical areas, they presented fairly similar profiles, indicating a potential common source. We concluded that PBDE levels in the farmed Chilean salmon are in the low average range of values published in the open literature.

Animals↗

Determination of DNA ploidy by fluorescence in situ hybridization (FISH) in hydatidiform moles: evaluation of FISH on isolated nuclei.

In the past 20 years, the diagnosis of hydatidiform moles has become more difficult because of the widespread use of early uterine evacuation. Differentiating hydropic degeneration, partial, and complete moles is important because of their different prognosis. However, clinical diagnosis is less obvious, and the pathologist has to separate the different entities on the basis of very subtle morphologic criteria. In difficult cases, ploidy may be determined by various methods, including fluorescence in situ hybridization (FISH) on routine histological sections from paraffin-embedded specimens. However, FISH analysis is often difficult because of the presence of numerous truncated nuclei. In this context, we have tested the advantages of FISH on isolated nuclei, a well-known variant of the technique that might be more sensitive. We reviewed 24 cases of products of abortion: hydropic degenerations, complete hydatidiform moles, partial moles, and nonmolar triploidies. After histological review, FISH on isolated nuclei proved conclusive in all cases. The results could be easily interpreted thanks to the reduced number of truncated nuclei. The percentage of cells with 2 signals was always >70% in the diploid cases and >60% in the triploid cases. In conclusion, this sensitive technique seems to be a valuable tool for the diagnosis of moles.

Cell Nucleus↗

Prey catching in the archer fish: does the fish use a learned correction for refraction?

An answer to the question of how the archer fish hits an aerial insect, despite the refraction of light at the surface of the water has not yet been found. The aims of the present studies are to find out: (1) whether the fish applies a learned correction with the virtual image as a point of reference; (2) whether deprivation of practice in squirting affects performance. For the first aim the accuracy of squirts was measured in 30 subjects. Contrary to suggestions from the literature, elevation failures were prominent but the frequencies of over- and under-squirting did not differ, which does not support the idea that the fishes applied a learned correction for refraction by using feedback from the efficacy of squirts. For the second aim, five experimental subjects were deprived of practice, whereas six control subjects got daily practice, during 6 months. The only significant difference, found thereafter, was that during the first session experimental subjects aimed more often before squirting than control subjects did, but hitting was not affected. A number of subjects developed abnormal mandibles which inevitably led to squirting too high. Our findings do not support the hypothesis that the archer fish uses learned corrections for refraction.

Journal Article↗

Preliminary study on puffer fish proteome-species identification of puffer fish by two-dimensional electrophoresis.

The aims of this work were to determine the differential characterization of the urea soluble protein components of puffer fish species and to establish a preliminary proteomic database using an immobilized pH gradient two-dimensional electrophoresis (2DE) technique. The puffer fish muscle proteins resolved into 171-260 spots in the 2DE gels, with a pI range of 3-10 and molecular mass range of 7.4-205.0 kDa, following Comassie blue staining. Puffer fish muscle proteins fell in the region with pI values of 3.5-7.0, and molecular masses of 7.4-45.0 kDa were well-resolved and were good for species comparison. The more acidic proteins of lower molecular masses showed species specific characteristics. Therefore, the species of puffer fish can be differentiated from the comparison of the characteristic 2DE protein patterns.

Animals↗

A comparison of techniques for preparing fish fillet for ICP-AES multielemental analysis and the microwave digestion of whole fish.

Four catfish fillet homogenate treatments before multielemental metal analysis by simultaneous inductively coupled plasma/atomic emission spectroscopy were compared in triplicate. These treatments were: nitric acid wet-ashing by Parr bomb digestion; nitric acid wet-ashing by microwave digestion; tetramethylammonium hydroxide/nitric acid wet digestion; and dry-ashing. The tetramethylammonium hydroxide/nitric acid method was imprecise (coefficients of variation > 20%). The dry-ashing method was fast and sensitive but had low recoveries of 50% for spiked Pb and Al and was not as precise as the Parr bomb or microwave treatments. The Parr bomb method was the most precise method but was less sensitive than the microwave method which had nearly the same precision. The microwave method was then adapted to homogenates of small whole fish < or = 3 cm in length. The whole fish homogenate required more vigorous digestion conditions, and addition of more acid after the evaporative step because of the presence of less oxidizable and acid-soluble components than fillet. The whole fish homogenate was also more heterogeneous than catfish fillet. A quality assurance protocol to demonstrate homogenate uniformity is essential. The use of a non-specialized microwave oven system allowed precise results for fillet and whole fish homogenates.

Animals↗

Studying meiosis: a review of FISH and M-FISH techniques used in the analysis of meiotic processes in humans.

It is well known that chromosome in situ hybridization allows the unequivocal identification of targeted human somatic chromosomes. Different fluorescent in situ hybridization (FISH) techniques have been developed throughout the years and, following the mitotic studies, meiotic analyses have been performed using these different techniques. The introduction of M-FISH techniques to the analysis of meiotic cells has allowed the study of meiotic processes for every individual human chromosome. In this paper, we review the different FISH and M-FISH techniques that have been used on human meiotic cells in both men and women.

Diploidy↗

Hydrodynamics of unsteady fish swimming and the effects of body size: comparing the flow fields of fish larvae and adults.

Zebra danios (Brachydanio rerio) swim in a burst-and-coast mode. Most swimming bouts consist of a single tail flick and a coasting phase, during which the fish keeps its body straight. When visualising the flow in a horizontal section through the wake, the effects of the flow regime become apparent in the structure of the wake. In a two-dimensional, medio-frontal view of the flow, larvae and adults shed two vortices at the tail during the burst phase. These vortices resemble a cross section through a large-core vortex ring: two vortex cores packed close together with the central flow directed away from the fish. This flow pattern can be observed in larvae (body length approximately 4 mm) at Reynolds numbers below 100 as well as in adult fish (body length approximately 35 mm) at Reynolds numbers above 1000. Larval vortices differ from those of adult zebra danios mainly in their relatively wider vortex cores (higher ratio of core radius to ring radius) and their lower vortex circulation. Both effects result from the increased importance of viscosity on larval flows. During the coasting phase, larval and adult flows again differ because of the changing importance of viscosity. The high viscosity of the water causes large vortical flows adjacent to the larva's body. These regions of high vorticity represent the huge body of water dragged along by the larva, and they cause the larva to stop almost immediately after thrust generation ceases. No such areas of high vorticity are visible adjacent to adult zebra danios performing a comparable swimming manoeuvre. The rapid decrease in vortex circulation and the severe reduction in the coasting distance due to viscous drag contribute to the high cost that larvae - unlike adult fish - face when using a burst-and-coast swimming style.

Animals↗