Application of FISH to detect DNA damage. DNA breakage detection-FISH (DBD-FISH).
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DNA Damage↗
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Classification and coding of commercial fishing injuries by work processes: an experience in the Danish fresh market fishing industry.
BACKGROUND: Work-related injuries in commercial fishing are of concern internationally. To better identify the causes of injury, this study coded occupational injuries by working processes in commercial fishing for fresh market fish. METHODS: A classification system of the work processes was developed by participation in fishing vessel trips where observations and video recordings of the work operations on board were collected. Subsequently the system was pilot tested using the Danish Maritime Authority injury reports. RESULTS: The developed classification system contains 17 main categories and up to 13 sub-categories of the work processes for each of the five different types of fishing. A total of 620 injury reports were reviewed and coded. Five percent (n = 33) of these were fatal injuries. The working processes were identified and coded according to the developed classification system for 553 (89%) injury reports: Danish seiner (n = 83), gill-netter (n = 122), beam trawler (n = 71), twin-trawler 2-T (n = 96), single/pair trawler 1-T (n = 181). Sixty-seven (11%) of the reports were unclassifiable due to lack of information. Preparing, shooting, and hauling of the gear and nets accounted for 50% of the injuries; they were most serious type of injuries such as fractures and sprains. Walking about the ship, in particular embarking and disembarking, climbing and descending ladders accounted for nearly one-fifth of the injuries. CONCLUSION: We found that the working processes related to working with the gear and nets vary greatly in the different fishing methods. Coding of the injuries to the specific working processes allows for targeted prevention efforts.
Accidents, Occupational↗
Fish optic nerve oligodendrocytes support axonal regeneration of fish and mammalian retinal ganglion cells.
Segments from adult fish and rat retinae were explanted on myelin-marker expressing oligodendrocytes derived from the regenerating goldfish optic nerve. Fish axons grew in high density and even rat retinal axons regenerated to considerable length on the surface of the fish oligodendrocytes, suggesting that this type of fish glia has axon-growth promoting surface components that exert their influence across species boundaries. One interesting surface component of the fish oligodendrocytes as demonstrated here is the E 587 antigen, which is related to the L1 family of cell adhesion molecules. In long term cocultures of oligodendrocytes and retinal axons, the fish glial cells were found to enwrap rat axons. This suggests that the oligodendrocytes of the regenerating goldfish optic nerve/tract may, despite striking differences, represent the equivalent to mammalian optic nerve oligodendrocytes.
Animals↗
DNA breakage detection-FISH (DBD-FISH) in human spermatozoa: technical variants evidence different structural features.
Non-irradiated and X-irradiated (80 Gy) human spermatozoa were processed for in situ DNA breakage detection-FISH (DBD-FISH) of the whole genome, following two alternative variations of the basic technique. In the first, cells were initially incubated in the alkaline unwinding solution for transformation of DNA breaks into single-stranded DNA (ssDNA) to be hybridized, followed by the lysing solutions for protein removal. In the second, incubation in the lysing solutions was carried out before the denaturation step. The first approach yielded two subpopulations. While most sperm nuclei were faintly labeled and had chromocenters, a small subpopulation was strongly and homogeneously labeled, due to extensive DNA breakage. X-ray exposure increased the surface and mean fluorescence intensity. Otherwise, when the denaturation step was performed after protein extraction, all sperm nuclei yielded strong and dispersed FISH signals. Protein removal allows access of the unwinding solution to the DNA, which has abundant alkali-labile sites, and thus gives rise to large areas of ssDNA that are labeled by FISH. X-ray exposure increased the dispersion of FISH signals but decreased their mean fluorescence intensity. A linear dose-response was generated using the second experimental variant, being 30 Gy the lowest dose for detecting induction of damage by X-rays in mature sperm chromatin. These results indicate that DBD-FISH is not only useful for in situ detection of DNA breakage but also for revealing structural features of chromatin.
DNA↗
Drinks like a fish: zebra fish (Danio rerio) as a behavior genetic model to study alcohol effects.
Zebra fish may be an ideal vertebrate model system for numerous human diseases with which the genetics and biological mechanisms of the disease may be studied. Zebra fish has been successfully used in developmental genetics, and recently, neurobiologists have also started to study this species. A potentially interesting target disease amenable for analysis with zebra fish is drug addiction, e.g. alcoholism. Although genetic tools to manipulate the genome of zebra fish are available, appropriate phenotypical testing methods are often lacking. In this paper, we describe basic behavioral tests to investigate the acute effects of alcohol on zebra fish. These behavioral paradigms will be useful for the genetic and biological analysis of acute and chronic drug effects as well as addiction. In addition to presenting findings for the acute effects of alcohol, we briefly describe our strategy for generating and screening mutants. We hope that our pilot work will facilitate the future development of behavioral tests and the use of zebra fish in the genetic analysis of the biological effects of drugs of abuse.
Aggression↗
The acute toxicity of surfactants on fish cells, Daphnia magna and fish-a comparative study.
There is a need to replace acute toxicity tests on fish (LC(50)) with more cost effective assays. The main objective of this study was to explore whether gill epithelial cells, hepatocytes and Daphnia magna could be used to predict acute toxicity of surfactants on fish. The acute toxicity of 10 synthetic surfactants (anionic, cationic, nonionic and zwitterionic) and two resin acids were determined on hepatocytes and gill epithelial cells from rainbow trout (Oncorhynchus mykiss), on Daphnia magna and on fish. Cell viability was measured with the fluorescent viability probe calcein-AM, immobilization was determined for Daphnia and 24-hr LC(50) for rainbow trout. The EC(50) values for the cellular tests were clearly higher than the corresponding values for Daphnia and fish, indicating that the cellular tests with the endpoint used are less sensitive than whole organisms. A combination of the EC(50) values for Daphnia and freshly isolated gill epithelial cells in suspension showed, however, a good correlation with acute toxicity on fish (r(2)=0.91 and slope=1.09). The combination seems to be a promising in vitro alternative to acute toxicity tests on fish (LC(50)), but a more exhaustive comparison, including a broad spectrum of chemicals should be made before the predictive value of the combined in vitro test can be evaluated.
Animal Testing Alternatives↗
[Infestation of larval trematodes from fresh-water fish and brackish-water fish in River Hyungsan, Kyungpook Province, Korea]
In order to determine the infestation patterns for metacercaria of digenetic trematodes, several kinds of fresh-water and brackish-water fish were caught in River Hyungsan, Kyungpook Province, Korea and examined from March to October in 1983. They were collected by netting, fishig with rod and line, or using the bait of crushed oil cake in transparent plastic bowls, and were dissected into the flesh, scales and fins with tail, and then examined for the presence and infestation density of larval trematodes. Of the 11 kinds of fish examined, the encysted larvae of Exorchis oviformis were found all of them, Metagonimus yokogawai from 9 kinds, Cyathocotyle orientalis, Metacercaria hasegawai and Metorchis orientalis from 6 kinds, and Clonorchis sinensis from 4 kinds in the present study. In the intensity of infestation with Clonorchis sinensis, the Korean shiner, Gnathopogon atromaculatus Nichols et Pope, was the most heavily infected species, the average number of metacercariae per gram of fish flesh being 45.5, followed by the striped shiner, Puntungia herzi Herzenstein 11.6, and Southern top-mouthed minnow, Pseudorasbora parva (Temminck et Shlegel), 9.7. The intensity of infestation with the Metagonimus larvae in 8 species of fresh-water fish was relatively low, and the average number per gram of flesh varied from 0.23 to 9.1. In comparison, the brackish-water fish were more heavily infected and the mean number of cysts per gram of flesh in the sweetfish, Plecoglossus altivelis, was 32.9. The results of this study generally indicate that Clonorchis sinensis and Metagonimus yokogawai infection may occur by consuming raw fresh-water fish caught in the river Hyungsan, Kyungpook Province, Korea.
Journal Article↗
Feeding ecology and food composition of the black carp Mylopharyngodon piceus and the grass carp Ctenopharyngodon idella inhabiting the fish pond of Al-Abbassa fish hatchery with emphasis given to vector snails.
Stomach contents of M. piceus and C. idella collected from earthen fish ponds at Al-Abbassa fish hatchery during different seasons were examined. Food items were analyzed by three methods; the numerical (N%) percentage of point assessment (P%) and frequency of occurrence (F%). A comparison of the different methods emphasized the importance of snails as the major food resources in the diet of M. piceus during all seasons. During winter, stomachs of the most fish samples collected from fish ponds were empty or had traces of food. The index of relative importance (I.R.I) indicated that snails were the major food category in the diet of M. piceus during summer, autumn and spring represented by 7331, 9696 and 11670 respectively. Plants were the main food item in the diet of C. idella during summer (4604), meanwhile they came 3rd in relative importance in the diet during autumn and spring. Artificial fish food came as the 2nd food item in the diet of M. piceus during summer (1398.9) and autumn (5896.3) but it completely absent in spring. It is considered the main food item in diet of C. idella during autumn (10384.3) and spring (6293.5) and being 3rd in summer (1395.5). The results showed that snails reached highest level in the diet of M. piceus during spring (about 80% of all diet, decreased during summer (about 70%) and reached lowest proportion in autumn (50%). Plants exhibited marked seasonal variation in abundance with maximum level during summer (about 41% of all diet decreased during spring (27%) and autumn (22%). Artificial fish food was the highest in the diet of M. piceus during autumn (39%), decreased in summer (22%) until disappeared during spring. It reached highest level in diet of C. idella during autumn (49%), decreased in spring (38%) and reached lowest proportion during summer (21 %).
Animals↗
A comparative analysis of somatolactin-related immunoreactivity in the pituitaries of four neopterygian fishes and one chondrostean fish: an immunohistochemical study.
An antiserum to cod somatolactin (SL) was used for immunohistochemical screening for the pars intermedia of two teleosts (Oreochromis mossambicus and Gymothorax meleagris), two holostean fishes (Lepisosteus osseus and Amia calva), and a chondrostean fish (Acipenser fulvescens) for SL-immunopositive (SL-IR) cells. As expected, a subset of the epithelial cells in the pars intermedia of O. mossambicus (tilapia) was immunopositive for SL, and the remainder of the epithelial cells was immunopositive for alpha-MSH-specific antiserum (alpha-MSH-IR). SL-IR was not detected in any of the epithelial cells in the pars intermedia of the moray eel G. meleagris. To determine whether SL-IR could be detected in nonteleost fishes, immunohistochemical analyses were done on the pituitaries of two holostean fishes and one chondrostean fish. In the pars intermedia of the gar, L. osseus, a subset of cells was immunopositive for alpha-MSH only. However, in the pars intermedia of the bowfin, A. calva, all of the epithelial cells indicated the presence of both SL and alpha-MSH. Finally, no SL-positive cells were detected in the pars intermedia of the sturgeon, A. fulvescens.
Animals↗
Formation of N-nitroso-N-methylurea in various samples of smoked/dried fish, fish sauce, seafoods, and ethnic fermented/pickled vegetables following incubation with nitrite under acidic conditions.
In continuation of our previous studies on N-nitroso-N-methylurea (NMU) formation in cured meats following incubation with nitrite at gastric pH, we extended the investigation to other foods mentioned in the title of this paper. The main objective was to determine whether these foods have the potential to form NMU at pH's that can be found in the human stomach. This was done by nitrosating an aliquot (5 g for fish sauce, 10 g for the others) of each with 7.25 microM to 1.59 mM levels of sodium nitrite for 2 h at room temperature at pH 0.8--1.5 and measuring the amounts of NMU formed. Of the samples tested, fish sauce formed 2--712 ng of NMU, followed in decreasing order by herring (<0.3--688 ng); dried anchovy, shrimp, and other fishes (<0.3--134 ng); crab and lobster paté (<0.3--342 ng); sardines (6--59 ng); oysters and mussels (11--31 ng); dried squid (3--47 ng); kimchi (7--107 ng); and Japanese pickled radish (<0.3--72 ng). Incorporation of 200-2000 ppm of ascorbic acid in the fish sauce and other foods, prior to nitrosation, appreciably inhibited such NMU formation. Although previous researchers in China reported NMU formation in nitrosated samples of fish sauce, this is the first reported formation of NMU upon nitrosation of the other foods mentioned above, and the first reported inhibition of such formation by added ascorbic acid.
Animals↗
Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish.
Striped jack nervous necrosis virus (SJNNV), a nodavirus, is the causative agent of viral nervous necrosis (VNN) in larval striped jack fish. In the present study, the SJNNV coat protein gene was sequenced and compared with that of four known insect nodaviruses and with four other fish nodaviruses causing VNN. The SJNNV coat protein gene was 1410 bases in length and contained a single ORF of 1023 bases encoding a protein of 340 amino acids. The sequence similarities between the coat protein gene of SJNNV and four insect nodaviruses were 28.6% or less at the nucleotide level and 10.6% or less at the amino acid level. A portion of the coat protein gene from four additional fish VNN viruses was amplified by PCR using primers designed for SJNNV and the amplified fragments (870-876 bases) were sequenced. The sequence similarities among SJNNV and the four VNN viruses were 75.8% or greater at the nucleotide level and 80.9% or greater at the amino acid level. In the fish nodaviruses a highly conserved region of 134 amino acids with sequence similarity of 92.5% or greater was detected. This conserved sequence was not found in the coat protein of insect nodaviruses. These results indicate that the fish nodaviruses that cause VNN are closely related to each other but are quite different from insect nodaviruses.
Amino Acid Sequence↗
[Evaluation of degree of freshness and value of fish and fish product t intake based on histamine and trimethylamine analysis].
The results of histamine and trimethylamine determination in three fish species (cod, herring, scomber) of various freshness are presented. In fresh fish the histamine contents were s follow: cod 0.00; scomber 1.00; and herring 0.50 mg/100 g. During storage time at room temperature (+18 degrees C), the value of the freshness indicators increased after 24 and 48 hours of storage. The highest increase was observed in scomber (21.30 and 34.00 mg/100 g), the lowest in cod (2.00 and 4.00 mg/100g). The trimethylamine value increased in different order than the histamine in the fish species. The highest TMA amount was observed in cod flesh--the lowest in scomber. According to these findings the freshness of some ready fish products was determined. The histamine level never exceeded 6 mg/100 g, always being below the "safety level" (= 10 mg/100g). The value of TMA never exceeded the limits considered for fresh fish products--15 mg N per 100 mg sample.
Animals↗
Biochemical and cellular effects of fish and fish oils.
The renewed interest in fish and fish oil has led to a re-examination of the biochemical and physiological basis of the preventive and therapeutic effects of fish and its components. Fatty acids are produced by a complex synthetic process. Unsaturated fatty acid derived from dietary sources are required for eicosanoid formation and are an integral part of cell wall and mitochondrial membranes. The beneficial role of fish oils is believed to stem from their being rich sources of omega-3 fatty acids. The dietary requirement of these fatty acids is a controversial issue but their essentiality for human health is being increasingly demonstrated and recognized. This review focusses on general aspects of lipid metabolism, biochemistry and physiological role of omega-3 fatty acids, and the unique characteristics of fatty acids found in fish. A subsequent review article will deal with metabolic and clinical aspects.
Animals↗
[Nutritional value of fish flours made in laboratory from fishes of the Nosy-Bé area].
Several types of fish meals (principally from no commercializable Carangues) are made in a laboratory. They are tested for general composition, amino-acid composition, vitamin and mineral compositions. Two fish meals from Carangues are very interesting: they are white muscle meal (rich in proteins of good quality, poor in lipids and slightly rich in minerals) and the meal from fish without head and guts (not so rich in proteins but slightly richer in minerals). These fish meals have a variable interest regarding mineral and vitamin contents. All of them have a lack of methionin and cystin which is comparable to the defficiency mentioned in other fish meals by various authors.
Amino Acids↗
[Extractability of lipids from fish and fish products].
The completness of the lipids removal from the fish muscles and fish products was investigated by making extraction in a filtering separating funnel (FSF) formerly proposed for determining lipids in oil-bearing seeds and cereals. The new modified procedure was contrasted against the standard one (accepted by the State Standards) used in separating and quantitative determination of the lipids content in fish by using the Socklet apparatus for extraction. The proposed variant of extraction was found to secure a higher yield of the lipids. The pattern of analysis used in determining the lipids content in vegetable products may well be used also in investigating animal products, such as, fish and fish products.
Animals↗
Metal levels in fish from the Savannah River: potential hazards to fish and other receptors.
Fish are ideal indicators of heavy metal contamination in aquatic systems because they occupy different trophic levels and are different sizes and ages. In this paper, we report concentrations of arsenic, cadmium, chromium, copper, lead, manganese, strontium(88) and mercury in the muscle of 11 species of fish from the Savannah River near the Savannah River Site. We test the hypotheses that there are no locational, species, or trophic-level differences in contaminant levels. There were significant species differences for all metals; higher-trophic-level fish generally had higher levels of arsenic, chromium, and copper. There were relatively few locational differences, and where there were such differences, they were small. The relationships between body weight and contaminant levels were generally positive, except for strontium, where there was a negative correlation for bowfin (Amia cal va), bass (Micropterus salmoides), yellow perch (Perca flavescens), and shellcracker (Lepomis microlophus) and no relationship for the other species. The levels of most metals were similar to, or lower than, those for the United States generally, and the levels of metals in fish from the Savannah River do not appear to pose a health threat to the fish themselves or to higher-order consumers, based on levels known to cause effects.
Animals↗
Heavy metal concentrations in marine fishes collected from fish culture sites in Hong Kong.
The levels of six heavy metals (Cd, Cr, Cu, Ni, Pb, and Zn) in different tissues of three species of cultured marine fishes (Epinephelus areolatus, Lutjanus russelli, and Sparus sarba) collected from three fish culture sites in Hong Kong were evaluated. Metal pollution problems in the fish culture sites were serious, as reflected by the high metal concentrations recorded in sea water, sediments, and the biomonitor Perna viridis. In general, tissues of all three species contained high concentrations of Zn and Cu, but much lower concentrations of Ni, Pb, Cd, and Cr. Similar pattern of heavy metal concentrations was observed in sea water, sediment, and P. viridis. Metal concentrations in various tissues varied greatly among species and among fish culture sites. Different tissues showed different capacity for accumulating heavy metals. Gonad of all three species contained high concentrations of Zn. On the other hand, liver seemed to be the primary organ for Cu accumulation. Overall, metal concentrations in the tissues of culture marine fishes were much lower than those in P. viridis. Despite high metal levels in sea water and sediments, concentrations of Cd, Cr, and Pb in edible tissues, including muscle and skin, did not exceed permissible levels recommended by the Hong Kong Government for human consumption.
Animals↗
Science, policy, stakeholders, and fish consumption advisories: developing a fish fact sheet for the Savannah River.
In recent years there has been a startling rise in the issuance of fish consumption advisories. Unfortunately, compliance by the public is often low. Low compliance can be due to a number of factors, including confusion over the meaning of advisories, conflicting advisories issued by different agencies, controversies involving health benefits versus the risks from consuming fish, and an unwillingness to act on the advisories because of personal beliefs. In some places, such as along the Savannah River, one state (South Carolina) had issued a consumption advisory while the other (Georgia) had not, although at present, both states now issue consumption advisories for the Savannah River. Herein we report on the development of a fish fact sheet to address the confusing and conflicting information available to the public about consuming fish from the Savannah River. The process involved interviewing fishers to ascertain fishing and consumption patterns, evaluating contaminant levels and exposure pathways, discussing common grounds for the provision of information, and consensus-building among different regulatory agencies (US Environmental Protection Agency, South Carolina Department of Health and Environmental Control, Georgia Department of Natural Resources) and the Department of Energy. Consensus, a key ingredient in solving many different types of "commons" problems, was aided by an outside organization, the Consortium for Risk Evaluation with Stakeholder Participation (CRESP). The initial role for CRESP was to offer scientific data as a basis for groups with different assumptions about risks to reach agreement on a regulatory response action. The process was an example of how credible science can be used to implement management and policies and provide a basis for consensus-building on difficult risk communication issues. The paper provides several lessons for improving the risk process from stakeholder conflicts, through risk assessment, to risk management. It also suggests that consensus-building and risk communication are continuing processes that involve assimilation of new information on contaminants and food-chain processes, state and federal law, public policy, and public response.
Animals↗