Single-Molecule Fluorescence In Situ Hybridization (FISH) of Circular RNA CDR1as.

PubMed

Kocks, Christine; Boltengagen, Anastasiya; Piwecka, Monika; Rybak-Wolf, Agnieszka; Rajewsky, Nikolaus

2018-01-01

Individual mRNA molecules can be imaged in fixed cells by hybridization with multiple, singly labeled oligonucleotide probes, followed by computational identification of fluorescent signals. This approach, called single-molecule RNA fluorescence in situ hybridization (smRNA FISH), allows subcellular localization and absolute quantification of RNA molecules in individual cells. Here, we describe a simple smRNA FISH protocol for two-color imaging of a circular RNA, CDR1as, simultaneously with an unrelated messenger RNA. The protocol can be adapted to circRNAs that coexist with overlapping, noncircular mRNA isoforms produced from the same genetic locus.

Telomeres and NextGen CO-FISH: Directional Genomic Hybridization (Telo-dGH™).

PubMed

McKenna, Miles J; Robinson, Erin; Goodwin, Edwin H; Cornforth, Michael N; Bailey, Susan M

2017-01-01

The cytogenomics-based methodology of Directional Genomic Hybridization (dGH™) emerged from the concept of strand-specific hybridization, first made possible by Chromosome Orientation FISH (CO-FISH), the utility of which was demonstrated in a variety of early applications, often involving telomeres. Similar to standard whole chromosome painting (FISH), dGH™ is capable of identifying inter-chromosomal rearrangements (translocations between chromosomes), but its distinctive strength stems from its ability to detect intra-chromosomal rearrangements (inversions within chromosomes), and to do so at higher resolution than previously possible. dGH™ brings together the strand specificity and directionality of CO-FISH with sophisticated bioinformatics-based oligonucleotide probe design to unique sequences. dGH™ serves not only as a powerful discovery tool-capable of interrogating the entire genome at the megabase level-it can also be used for high-resolution targeted detection of known inversions, a valuable attribute in both research and clinical settings. Detection of chromosomal inversions, particularly small ones, poses a formidable challenge for more traditional cytogenetic approaches, especially when they occur near the ends or telomeric regions. Here, we describe Telo-dGH™, a strand-specific scheme that utilizes dGH™ in combination with telomere CO-FISH to differentiate between terminal exchange events, specifically terminal inversions, and an altogether different form of genetic recombination that often occurs near the telomere, namely sister chromatid exchange (SCE).

Interspecific hybridization causes long-term phylogenetic discordance between nuclear and mitochondrial genomes in freshwater fishes.

PubMed

Wallis, Graham P; Cameron-Christie, Sophia R; Kennedy, Hannah L; Palmer, Gemma; Sanders, Tessa R; Winter, David J

2017-06-01

Classification, phylogeography and the testing of evolutionary hypotheses rely on correct estimation of species phylogeny. Early molecular phylogenies often relied on mtDNA alone, which acts as a single linkage group with one history. Over the last decade, the use of multiple nuclear sequences has often revealed conflict among gene trees. This observation can be attributed to hybridization, lineage sorting, paralogy or selection. Here, we use 54 groups of fishes from 48 studies to estimate the degree of concordance between mitochondrial and nuclear gene trees in two ecological grades of fishes: marine and freshwater. We test the hypothesis that freshwater fish phylogenies should, on average, show more discordance because of their higher propensity for hybridization in the past. In keeping with this idea, concordance between mitochondrial and nuclear gene trees (as measured by proportion of components shared) is on average 50% higher in marine fishes. We discuss why this difference almost certainly results from introgression caused by greater historical hybridization among lineages in freshwater groups, and further emphasize the need to use multiple nuclear genes, and identify conflict among them, in estimation of species phylogeny. © 2017 John Wiley & Sons Ltd.

Detection of a complex translocation using fluorescent in situ hybridization (FISH)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rosen, B.A.; Abuelo, D.N.; Mark, H.F.

1994-09-01

The use of fluorescent in situ hybridization (FISH) allowed the detection of a complex 3-way translocation in a patient with multiple congenital malformations and mental retardation. The patient was a 10-year-old girl with mental retardation, seizures, repaired cleft palate, esotropia, epicanthal folds, broad nasal bridge, upward slanting palpebral fissures, single transverse palmar crease, brachydactyly, hypoplastic nails, ectrodactyly between the third and fourth right toes, and hypoplasia of the left third toe. Chromosome analysis performed at birth was reported as normal. We performed high resolution banding analysis which revealed an apparently balanced translocation between chromosomes 2 and 9. However, because ofmore » her multiple abnormalities, further studies were ordered. Fluorescent in situ hybridization (FISH) using chromosome painting probes revealed a karyotype of 46,XX,t(2;8;9) (2pter{yields}q31::8q21.2{yields}8qter; 8pter{yields}q21.2::2q31{yields}q34::9q34{yields}qter; 9pter{yields}q34::2q34{yields}qter). The 3-way translocation appears to be de novo, as neither parent is a translocation carrier. This case illustrates the importance of using FISH to further investigate cases of apparently balanced translocations in the presence of phenotypic abnormalities and/or mental retardation.« less

Recent advances in rice genome and chromosome structure research by fluorescence in situ hybridization (FISH).

PubMed

Ohmido, Nobuko; Fukui, Kiichi; Kinoshita, Toshiro

2010-01-01

Fluorescence in situ hybridization (FISH) is an effective method for the physical mapping of genes and repetitive DNA sequences on chromosomes. Physical mapping of unique nucleotide sequences on specific rice chromosome regions was performed using a combination of chromosome identification and highly sensitive FISH. Increases in the detection sensitivity of smaller DNA sequences and improvements in spatial resolution have ushered in a new phase in FISH technology. Thus, it is now possible to perform in situ hybridization on somatic chromosomes, pachytene chromosomes, and even on extended DNA fibers (EDFs). Pachytene-FISH allows the integration of genetic linkage maps and quantitative chromosome maps. Visualization methods using FISH can reveal the spatial organization of the centromere, heterochromatin/euchromatin, and the terminal structures of rice chromosomes. Furthermore, EDF-FISH and the DNA combing technique can resolve a spatial distance of 1 kb between adjacent DNA sequences, and the detection of even a 300-bp target is now feasible. The copy numbers of various repetitive sequences and the sizes of various DNA molecules were quantitatively measured using the molecular combing technique. This review describes the significance of these advances in molecular cytology in rice and discusses future applications in plant studies using visualization techniques.

Intragenus (Campylomormyrus) and intergenus hybrids in mormyrid fish: Physiological and histological investigations of the electric organ ontogeny.

PubMed

Kirschbaum, Frank; Nguyen, Linh; Baumgartner, Stephanie; Chi, Hiu Wan Linda; Wolfart, Rene; Elarbani, Khouloud; Eppenstein, Hari; Korniienko, Yevheniia; Guido-Böhm, Lilian; Mamonekene, Victor; Vater, Marianne; Tiedemann, Ralph

2016-10-01

African weakly electric mormyrid fish show a high diversity of their electric organ discharge (EOD) both across and within genera. Thanks to a recently developed technique of artificial reproduction in mormyrid fish, we were able to perform hybridizations between different genera and within one genus (Campylomormyrus). The hybrids of intergenus hybridizations exhibited different degrees of reduced survival related to the phylogenetic distance of the parent species: hybrids of the crosses between C. rhynchophorus and its sister genus Gnathonemus survived and developed normally. Hybrids between C. rhynchophorus and a Mormyrus species (a more basal clade compared to Campylomormyrus s) survived up to 42days and developed many malformations, e.g., at the level of the unpaired fins. Hybrids between C. numenius and Hippopotamyrus pictus (a derived clade, only distantly related to Campylomormyrus) only survived for two days during embryological development. Eight different hybrid combinations among five Campylomormyrus species (C. tamandua, C. compressirostris, C. tshokwe, C. rhynchophorus, C. numenius) were performed. The aim of the hybridizations was to combine species with (1) either caudal or rostral position of the main stalk innervating the electrocytes in the electric organ and (2) short, median or long duration of their EOD. The hybrids, though they are still juveniles, show very interesting features concerning electrocyte geometry as well as EOD form and duration: the caudal position of the stalk is prevailing over the rostral position, and the penetration of the stalk is dominant over the non-penetrating feature (in the Campylomormyrus hybrids); in the hybrid between C. rhynchophorus and Gnathonemus petersii it is the opposite. When crossing species with long and short EODs, it is always the long duration EOD that is expressed in the hybrids. The F1-Hybrids of the cross C. tamandua×C. compressirostris are fertile: viable F2-fish could be obtained with artificial

Design of Hybrid Solar and Wind Energy Harvester for Fishing Boat

NASA Astrophysics Data System (ADS)

Banjarnahor, D. A.; Hanifan, M.; Budi, E. M.

2017-07-01

In southern beach of West Java, Indonesia, there are many villagers live as fishermen. They use small boats for fishing, in one to three days. Therefore, they need a fish preservation system. Fortunately, the area has high potential of solar and wind energy. This paper presents the design of a hybrid solar and wind energy harvester to power a refrigerator in the fishing boat. The refrigerator should keep the fish in 2 - 4 °C. The energy needed is 720 Wh daily. In the area, the daily average wind velocity is 4.27 m/s and the sun irradiation is 672 W/m2. The design combined two 100W solar panels and a 300W wind turbine. The testing showed that the solar panels can harvest 815 - 817 Wh of energy, while the wind turbine can harvest 43 - 62 Wh of energy daily. Therefore, the system can fulfil the energy requirement in fishing boat, although the solar panels were more dominant. To install the wind turbine on the fishing-boat, a computational design had been conducted. The boat hydrostatic dimension was measured to determine its stability condition. To reach a stable equilibrium condition, the wind turbine should be installed no more than 1.7 m of height.

FISH-Based Markers Enable Identification of Chromosomes Derived From Tetraploid Thinopyrum elongatum in Hybrid Lines.

PubMed

Li, Daiyan; Li, Tinghui; Wu, Yanli; Zhang, Xiaohui; Zhu, Wei; Wang, Yi; Zeng, Jian; Xu, Lili; Fan, Xing; Sha, Lina; Zhang, Haiqin; Zhou, Yonghong; Kang, Houyang

2018-01-01

Tetraploid Thinopyrum elongatum , which has superior abiotic stress tolerance characteristics, and exhibits resistance to stripe rust, powdery mildew, and Fusarium head blight, is a wild relative of wheat and a promising source of novel genes for wheat improvement. Currently, a high-resolution Fluorescence in situ hybridization (FISH) karyotype of tetraploid Th. elongatum is not available. To develop chromosome-specific FISH-based markers, the hexaploid Trititrigia 8801 and two accessions of tetraploid Th. elongatum were characterized by different repetitive sequences probes. We found that all E-genome chromosomes could be unambiguously identified using a combination of pSc119.2, pTa535, pTa71, and pTa713 repeats, and the E-genome chromosomes of the wild accessions and the partial amphiploid failed to exhibit any significant variation in the probe hybridization patterns. To verify the validation of these markers, the chromosome constitution of eight wheat- Th. elongatum hybrid derivatives were analyzed. We revealed that these probes could quickly detect wheat and tetraploid Th. elongatum chromosomes in hybrid lines. K16-712-1-2 was a 1E (1D) chromosome substitution line, K16-681-4 was a 2E disomic chromosome addition line, K16-562-3 was a 3E, 4E (3D, 4D) chromosome substitution line, K15-1033-8-2 contained one 4E, two 5E, and one 4ES⋅1DL Robertsonian translocation chromosome, and four other lines carried monosomic 4E, 5E, 6E, and 7E chromosome, respectively. Furthermore, the E-genome specific molecular markers analysis corresponded perfectly with the FISH results. The developed FISH markers will facilitate rapid identification of tetraploid Th. elongatum chromosomes in wheat improvement programs and allow appropriate alien chromosome transfer.

High-resolution mapping reveals hundreds of genetic incompatibilities in hybridizing fish species.

PubMed

Schumer, Molly; Cui, Rongfeng; Powell, Daniel L; Dresner, Rebecca; Rosenthal, Gil G; Andolfatto, Peter

2014-06-04

Hybridization is increasingly being recognized as a common process in both animal and plant species. Negative epistatic interactions between genes from different parental genomes decrease the fitness of hybrids and can limit gene flow between species. However, little is known about the number and genome-wide distribution of genetic incompatibilities separating species. To detect interacting genes, we perform a high-resolution genome scan for linkage disequilibrium between unlinked genomic regions in naturally occurring hybrid populations of swordtail fish. We estimate that hundreds of pairs of genomic regions contribute to reproductive isolation between these species, despite them being recently diverged. Many of these incompatibilities are likely the result of natural or sexual selection on hybrids, since intrinsic isolation is known to be weak. Patterns of genomic divergence at these regions imply that genetic incompatibilities play a significant role in limiting gene flow even in young species.

Influence of environment and mitochondrial heritage on the ecological characteristics of fish in a hybrid zone.

PubMed

Stolzenberg, Nicolas; Nguyen The, Bénédicte; Salducci, Marie Dominique; Cavalli, Laurent

2009-06-18

Ecological characteristics (growth, morphology, reproduction) arise from the interaction between environmental factors and genetics. Genetic analysis of individuals' life history traits might be used to improve our understanding of mechanisms that form and maintain a hybrid zone. A fish hybrid zone was used to characterize the process of natural selection. Data were collected during two reproductive periods (2001 and 2002) and 1117 individuals (nase, Chondrostama nasus nasus, sofie C. toxostoma toxostoma and hybrids) were sampled. Reproductive dates of the two parental species overlapped at sympatric sites. The nase had an earlier reproductive period than the sofie; males had longer reproductive periods for both species. Hybridisation between female nase and male sofie was the most likely. Hybrids had a reproductive period similar to the inherited parental mitochondrial type. Growth and reproductive information from different environments has been synthesised following a bayesian approach of the von Bertalanffy model. Hybrid life history traits appear to link with maternal heritage. Hybrid size from the age of two and size at first maturity appeared to be closer to the size of the maternal origin species (nase or sofie). Median growth rates for hybrids were similar and intermediate between those of the parental species. We observed variable life history traits for hybrids and pure forms in the different parts of the hybrid zone. Geometrical analysis of the hybrid fish shape gave evidence of two main morphologies with a link to maternal heritage. Selective mating seemed to be the underlying process which, with mitochondrial heritage, could explain the evolution of the studied hybrid zone. More generally, we showed the importance of studies on hybrid zones and specifically the study of individuals' ecological characteristics, to improve our understanding of speciation.

Identification and characterization of karyotype in Passiflora hybrids using FISH and GISH.

PubMed

Silva, Gonçalo Santos; Souza, Margarete Magalhães; de Melo, Cláusio Antônio Ferreira; Urdampilleta, Juan Domingo; Forni-Martins, Eliana Regina

2018-04-27

A great interest exists in the production of hybrid plants of the genus Passiflora given the beauty and exotic features of its flowers which have ornamental value. Hybrid paternity confirmation is therefore important for assuring germplasm origin, and is typically carried out by molecular marker segregation. The aim of this study was to karyotypically characterize the chromosome heritance patterns of the progeny resultant from a cross of P. gardneri and P. gibertii using classical cytogenetics, chromosome banding, and molecular cytogenetics. All analyzed genotypes showed the same diploid chromosome number as the genitor species: 2n = 18. Classical and CMA 3 and DAPI staining allowed for chromosome counting and satellite identification (secondary constrictions). Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used to characterize subgenomes by either identifying rDNA-specific genome patterns or parental genomes, respectively. The heritance of chromosomal markers presenting rDNA sites from each parent for genome identification confirmed that all obtained plants were hybrids. These results will improve breeding programs involving the species of this genus. Apart from confirming hybridization, GISH allowed the visualization of recombination between the homeologous chromosome and the introgression of sequences of interest.

Fully Automated Fluorescent in situ Hybridization (FISH) Staining and Digital Analysis of HER2 in Breast Cancer: A Validation Study

PubMed Central

van der Logt, Elise M. J.; Kuperus, Deborah A. J.; van Setten, Jan W.; van den Heuvel, Marius C.; Boers, James. E.; Schuuring, Ed; Kibbelaar, Robby E.

2015-01-01

HER2 assessment is routinely used to select patients with invasive breast cancer that might benefit from HER2-targeted therapy. The aim of this study was to validate a fully automated in situ hybridization (ISH) procedure that combines the automated Leica HER2 fluorescent ISH system for Bond with supervised automated analysis with the Visia imaging D-Sight digital imaging platform. HER2 assessment was performed on 328 formalin-fixed/paraffin-embedded invasive breast cancer tumors on tissue microarrays (TMA) and 100 (50 selected IHC 2+ and 50 random IHC scores) full-sized slides of resections/biopsies obtained for diagnostic purposes previously. For digital analysis slides were pre-screened at 20x and 100x magnification for all fluorescent signals and supervised-automated scoring was performed on at least two pictures (in total at least 20 nuclei were counted) with the D-Sight HER2 FISH analysis module by two observers independently. Results were compared to data obtained previously with the manual Abbott FISH test. The overall agreement with Abbott FISH data among TMA samples and 50 selected IHC 2+ cases was 98.8% (κ = 0.94) and 93.8% (κ = 0.88), respectively. The results of 50 additionally tested unselected IHC cases were concordant with previously obtained IHC and/or FISH data. The combination of the Leica FISH system with the D-Sight digital imaging platform is a feasible method for HER2 assessment in routine clinical practice for patients with invasive breast cancer. PMID:25844540

A Winner Determination Algorithm for Combinatorial Auctions Based on Hybrid Artificial Fish Swarm Algorithm

NASA Astrophysics Data System (ADS)

Zheng, Genrang; Lin, ZhengChun

The problem of winner determination in combinatorial auctions is a hotspot electronic business, and a NP hard problem. A Hybrid Artificial Fish Swarm Algorithm(HAFSA), which is combined with First Suite Heuristic Algorithm (FSHA) and Artificial Fish Swarm Algorithm (AFSA), is proposed to solve the problem after probing it base on the theories of AFSA. Experiment results show that the HAFSA is a rapidly and efficient algorithm for The problem of winner determining. Compared with Ant colony Optimization Algorithm, it has a good performance with broad and prosperous application.

A patient with myelodysplastic syndrome studied by GTG-banding and fluorescent in situ hybridization (FISH).

PubMed

Mark, H F; Mark, Y; Sotomayor, E; Sambandam, S

1998-01-01

Molecular cytogenetics using fluorescent in situ hybridization (FISH) is an extremely useful adjunct technique to conventional cytogenetics via GTG-banding. The present paper illustrates the utility of FISH by describing a patient with myelodysplastic syndrome (MDS) who was initially studied using GTG-banding and whose bone marrow was found to be populated with hyperdiploid cells. FISH was used to delineate the numerical and structural chromosomal abnormalities. It revealed the presence of trisomy 8 and determined that the previously unidentifiable marker chromosome was of chromosome 10 origin. Although trisomy 8 is a frequent finding in MDS, the structural chromosomal abnormality of chromosome 10 as reported here is not a common finding.

Hybrid striped bass feeds based on fish oil, beef tallow, and eicosapentaenoic acid/docosahexaenoic acid supplements: Insight regarding fish oil sparing and demand for -3 long-chain polyunsaturated fatty acids.

PubMed

Bowzer, J; Jackson, C; Trushenski, J

2016-03-01

Previous research suggests that saturated (SFA) and monounsaturated fatty acid (MUFA) rich lipids, including beef tallow, can make utilization or diet-to-tissue transfer of long-chain polyunsaturated fatty acids (LC-PUFA) more efficient. We hypothesized that using beef tallow as an alternative to fish oil may effectively reduce the LC-PUFA demand of hybrid striped bass × and allow for greater fish oil sparing. Accordingly, we evaluated growth performance and tissue fatty acid profiles of juvenile fish (23.7 ± 0.3 g) fed diets containing menhaden fish oil (considered an ideal source of LC-PUFA for this taxon), beef tallow (BEEF ONLY), or beef tallow amended with purified sources of eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) to achieve levels corresponding to 50 or 100% of those observed in the FISH ONLY feed. Diets were randomly assigned to quadruplicate tanks of fish ( = 4; 10 fish/tank), and fish were fed assigned diets to apparent satiation once daily for 10 wk. Survival (98-100%) was equivalent among treatments, but weight gain (117-180%), specific growth rate (1.1-1.5% BW/d), feed intake (1.4-1.8% BW/d), thermal growth coefficient (0.50-0.70), and feed conversion ratio (FCR; 1.1-1.4, DM basis) varied. Except for FCR, no differences were observed between the FISH ONLY and BEEF ONLY treatments, but performance was generally numerically superior among fish fed the diets containing beef tallow supplemented with DHA at the 100% or both EPA and DHA at the 50% or 100% level. Tissue fatty acid composition was significantly distorted in favor among fish fed the beef tallow-based feeds; however, profile distortion was most overt in peripheral tissues. Results suggest that beef tallow may be used as a primary lipid source in practical diets for hybrid striped bass, but performance may be improved by supplementation with LC-PUFA, particularly DHA. Furthermore, our results suggest that -3 LC-PUFA requirements reported for hybrid striped bass may not be

Does interspecies hybridization affect the host specificity of parasites in cyprinid fish?

PubMed

Simková, Andrea; Dávidová, Martina; Papoušek, Ivo; Vetešník, Lukáš

2013-04-12

Host specificity varies among parasite species. Some parasites are strictly host-specific, others show a specificity for congeneric or non-congeneric phylogenetically related host species, whilst some others are non-specific (generalists). Two cyprinids, Cyprinus carpio and Carassius gibelio, plus their respective hybrids were investigated for metazoan parasites. The aim of this study was to analyze whether interspecies hybridization affects host specificity. The different degrees of host specificity within a phylogenetic framework were taken into consideration (i.e. strict specialist, intermediate specialist, and intermediate generalist). Fish were collected during harvesting the pond and identified using meristic traits and molecular markers. Metazoan parasite species were collected. Host specificity of parasites was determined using the following classification: strict specialist, intermediate specialist, intermediate generalist and generalist. Parasite species richness was compared between parental species and their hybrids. The effect of host species on abundance of parasites differing in host specificity was tested. Hybrids harbored more different parasite species but their total parasite abundance was lower in comparison with parental species. Interspecies hybridization affected the host specificity of ecto- and endoparasites. Parasite species exhibiting different degrees of host specificity for C. carpio and C. gibelio were also present in hybrids. The abundance of strict specialists of C. carpio was significantly higher in parental species than in hybrids. Intermediate generalists parasitizing C. carpio and C. gibelio as two phylogenetically closely related host species preferentially infected C. gibelio when compared to C. carpio, based on prevalence and maximum intensity of infection. Hybrids were less infected by intermediate generalists when compared to C. gibelio. This finding does not support strict co-adaptation between host and parasite genotypes

Visualization of sporopollenin-containing pathogenic green micro-alga Prototheca wickerhamii by fluorescent in situ hybridization (FISH).

PubMed

Ueno, Ryohei

2009-04-01

Fluorescent in situ hybridization (FISH) using taxon-specific, rRNA-targeted oligonucleotide probes is one of the most powerful tools for the rapid identification of harmful microorganisms. However, eukaryotic algal cells do not always allow FISH probes to permeate over their cell walls. Members of the pathogenic micro-algal genus Prototheca are characterized by their distinctive cell-wall component, sporopollenin, an extremely tough biopolymer that resists acid and alkaline hydrolysis, enzyme attack, and acetolysis. To our knowledge, there has been no report of the successful permeation by the oligonucleotide probes over the cell walls of unicellular green micro-algae, which contain sporopollenin. The DNA probes passed through the cell wall of Prototheca wickerhamii after treating the algal cells with cetyltrimethylammonium bromide (CTAB). Most cells in the middle logarithmic growth phase culture fluoresced when hybridized with the rRNA-targeted universal probe for eukaryotes, though individual cells included in this culture differed in the level of cell-wall vulnerability to attack by the polysaccharide-degrading enzyme, thus reflecting the different stages of the life cycle. This is the first report regarding the visualization of sporopollenin-containing, green micro-algal cells by FISH.

Visualization of mcr mRNA in a methanogen by fluorescence in situ hybridization with an oligonucleotide probe and two-pass tyramide signal amplification (two-pass TSA-FISH).

PubMed

Kubota, Kengo; Ohashi, Akiyoshi; Imachi, Hiroyuki; Harada, Hideki

2006-09-01

Two-pass tyramide signal amplification-fluorescence in situ hybridization (two-pass TSA-FISH) with a horseradish peroxidase (HRP)-labeled oligonucleotide probe was applied to detect prokaryotic mRNA. In this study, mRNA of a key enzyme for methanogenesis, methyl coenzyme M reductase (mcr), in Methanococcus vannielii was targeted. Applicability of mRNA-targeted probes to in situ hybridization was verified by Clone-FISH. It was observed that sensitivity of two-pass TSA-FISH was significantly higher than that of TSA-FISH, which was further increased by the addition of dextran sulphate in TSA working solution. Signals from two-pass TSA-FISH were more reliable compared to the weak, spotty signals yielded by TSA-FISH.

ASSESSING FISH TISSUE CONTAMINATION ON A REGIONAL SCALE

EPA Science Inventory

The selection of target fish species for assessing the extent of fish tissue contaminants is a critical consideration in regional stream surveys such as the Environmental Monitoring and Assessment Program (EMAP). The ideal species would be widely distributed and common, bioaccumu...

Genome analysis of 7 Kengyilia (Triticeae Poaceae) species with FISH and GISH

USDA-ARS?s Scientific Manuscript database

Genome composition of and genetic relationships among seven Kengyilia species were assessed using a technique of sequential FISH (fluorescence in situ hybridization) and GISH (genomic in situ hybridization). Five of these 7 species, K. kokonorica, K. rigidula, K. hirsula, K. grandiglumis, and K. th...

A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.

PubMed

Zeller, Perrine; Ploux, Olivier; Méjean, Annick

2016-03-01

Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

In vitro formation and thermal transition of novel hybrid fibrils from type I fish scale collagen and type I porcine collagen

NASA Astrophysics Data System (ADS)

Chen, Song; Ikoma, Toshiyuki; Ogawa, Nobuhiro; Migita, Satoshi; Kobayashi, Hisatoshi; Hanagata, Nobutaka

2010-06-01

Novel type I collagen hybrid fibrils were fabricated by neutralizing a mixture of type I fish scale collagen solution and type I porcine collagen solution with a phosphate buffer saline at 28 °C. Their structure was discussed in terms of the volume ratio of fish/porcine collagen solution. Scanning electron and atomic force micrographs showed that the diameter of collagen fibrils derived from the collagen mixture was larger than those derived from each collagen, and all resultant fibrils exhibited a typical D-periodic unit of ~67 nm, irrespective of volume ratio of both collagens. Differential scanning calorimetry revealed only one endothermic peak for the fibrils derived from collagen mixture or from each collagen solution, indicating that the resultant collagen fibrils were hybrids of type I fish scale collagen and type I porcine collagen.

Diagnostic and Prognostic Utility of Fluorescence In situ Hybridization (FISH) Analysis in Acute Myeloid Leukemia.

PubMed

Gonzales, Patrick R; Mikhail, Fady M

2017-12-01

Acute myeloid leukemia (AML) is a hematologic neoplasia consisting of incompletely differentiated hematopoietic cells of the myeloid lineage that proliferate in the bone marrow, blood, and/or other tissues. Clinical implementation of fluorescence in situ hybridization (FISH) in cytogenetic laboratories allows for high-resolution analysis of recurrent structural chromosomal rearrangements specific to AML, especially in AML with normal karyotypes, which comprises approximately 33-50% of AML-positive specimens. Here, we review the use of several FISH probe strategies in the diagnosis of AML. We also review the standards and guidelines currently in place for use by clinical cytogenetic laboratories in the evaluation of AML. Updated standards and guidelines from the WHO, ACMG, and NCCN have further defined clinically significant, recurring cytogenetic anomalies in AML that are detectable by FISH. FISH continues to be a powerful technique in the diagnosis of AML, with higher resolution than conventional cytogenetic analysis, rapid turnaround time, and a considerable diagnostic and prognostic utility.

Combining fluorescent in situ hybridization data with ISS staging improves risk assessment in myeloma: an International Myeloma Working Group collaborative project.

PubMed

Avet-Loiseau, H; Durie, B G M; Cavo, M; Attal, M; Gutierrez, N; Haessler, J; Goldschmidt, H; Hajek, R; Lee, J H; Sezer, O; Barlogie, B; Crowley, J; Fonseca, R; Testoni, N; Ross, F; Rajkumar, S V; Sonneveld, P; Lahuerta, J; Moreau, P; Morgan, G

2013-03-01

The combination of serum β2-microglobulin and albumin levels has been shown to be highly prognostic in myeloma as the International Staging System (ISS). The aim of this study was to assess the independent contributions of ISS stage and cytogenetic abnormalities in predicting outcomes. A retrospective analysis of international studies looking at both ISS and cytogenetic abnormalities was performed in order to assess the potential role of combining ISS stage and cytogenetics to predict survival. This international effort used the International Myeloma Working Group database of 12 137 patients treated worldwide for myeloma at diagnosis, of whom 2309 had cytogenetic studies and 5387 had analyses by fluorescent in situ hybridization (iFISH). Comprehensive analyses used 2642 patients with sufficient iFISH data available. Using the comprehensive iFISH data, combining both t(4;14) and deletion (17p), along with ISS stage, significantly improved the prognostic assessment in terms of progression-free survival and overall survival. The additional impact of patient age and use of high-dose therapy was also demonstrated. In conclusion, the combination of iFISH data with ISS staging significantly improves risk assessment in myeloma.

In situ DNA hybridized chain reaction (FISH-HCR) as a better method for quantification of bacteria and archaea within marine sediment

NASA Astrophysics Data System (ADS)

Buongiorno, J.; Lloyd, K. G.; Shumaker, A.; Schippers, A.; Webster, G.; Weightman, A.; Turner, S.

2015-12-01

Nearly 75% of the Earth's surface is covered by marine sediment that is home to an estimated 2.9 x 1029 microbial cells. A substantial impediment to understanding the abundance and distribution of cells within marine sediment is the lack of a consistent and reliable method for their taxon-specific quantification. Catalyzed reporter fluorescent in situ hybridization (CARD-FISH) provides taxon-specific enumeration, but this process requires passing a large enzyme through cell membranes, decreasing its precision relative to general cell counts using a small DNA stain. In 2015, Yamaguchi et al. developed FISH hybridization chain reaction (FISH-HCR) as an in situ whole cell detection method for environmental microorganisms. FISH-HCR amplifies the fluorescent signal, as does CARD-FISH, but it allows for milder cell permeation methods that might prevent yield loss. To compare FISH-HCR to CARD-FISH, we examined bacteria and archaea cell counts within two sediment cores, Lille Belt (~78 meters deep) and Landsort Deep (90 meters deep), which were retrieved from the Baltic Sea Basin during IODP Expedition 347. Preliminary analysis shows that CARD-FISH counts are below the quantification limit for most depths across both cores. By contrast, quantification of cells was possible with FISH-HCR in all examined depths. When quantification with CARD-FISH was above the limit of detection, counts with FISH-HCR were up to 11 fold higher for Bacteria and 3 fold higher for Archaea from the same sediment sample. Further, FISH-HCR counts follow the trends of on board counts nicely, indicating that FISH-HCR may better reflect the cellular abundance within marine sediment than other quantification methods, including qPCR. Using FISH-HCR, we found that archaeal cell counts were on average greater than bacterial cell counts, but within the same order of magnitude.

Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cui, Yi; Hu, Dehong; Markillie, Lye Meng

Quantitative gene expression analysis in intact single cells can be achieved using single molecule- based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple FISH sub-probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific binding of sub-probes and tissue autofluorescence, limiting its accuracy. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on blinking frequencies of photoswitchable dyes. This fluctuation localization imaging-based FISH (fliFISH) uses blinking frequency patterns, emitted frommore » a transcript bound to multiple sub-probes, which are distinct from blinking patterns emitted from partial or nonspecifically bound sub-probes and autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2, and their ratio (Nkx2-2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct blinking frequency patterns, laying the foundation for reliable single-cell transcriptomics.« less

Lake Ontario benthic prey fish assessment, 2016

USGS Publications Warehouse

Weidel, Brian C.; Walsh, Maureen; Holden, Jeremy P.; Connerton, Michael J.

2017-01-01

Benthic prey fishes are a critical component of the Lake Ontario food web, serving as energy vectors from benthic invertebrates to native and introduced piscivores. Beginning in 1978, Lake Ontario benthic prey fishes were assessed using bottom trawls collected from the lake’s south shore (depth range: 8 – 150 m). Historically, the survey targeted the then dominant species, Slimy Sculpin, however in 2015, the Benthic Prey Fish Survey was cooperatively expanded to a whole-lake survey, to address resource management information needs related to Round Goby, Deepwater Sculpin, and nearshore native fishes. In 2016, 142 trawls were collected at 18 transects, and spanned depths from 6 – 225 m. Trawl catches indicated the benthic and demersal prey fish community was dominated by Round Goby, however the proportional importance of native Deepwater Sculpin is increasing. Species-specific assessments found lake-wide Round Goby density (~600 fish per hectare) was slightly lower in 2016 relative to 2015. Deepwater Sculpin density has generally increased since 2004. In 2016 their estimated density was greater than 100 fish per hectare. Slimy Sculpin density (15 fish/ha) was similar to the past 3 years. Catches of juvenile Slimy Sculpin continue to be low relative to historic catches and the timing of their decline coincides with the proliferation of Round Goby. Additionally, we found a strong negative relationship between trawl catches of Round Goby and near-shore native benthic and demersal fishes such as Trout-perch, Johnny Darter and Spottail Shiner. The introduction of Round Goby and the reappearance of native Deepwater Sculpin have shaped the Lake Ontario benthic prey fish community.

Lake Ontario benthic prey fish assessment, 2015

USGS Publications Warehouse

Weidel, Brian C.; Walsh, Maureen; Holden, Jeremy P.; Connerton, Michael J.

2016-01-01

Benthic prey fishes are a critical component of the Lake Ontario food web, serving as energy vectors from benthic invertebrates to native and introduced piscivores. Since the late 1970’s, Lake Ontario benthic prey fish status was primarily assessed using bottom trawl observations confined to the lake’s south shore, in waters from 8 – 150 m (26 – 492 ft). In 2015, the Benthic Prey Fish Survey was cooperatively adjusted and expanded to address resource management information needs including lake-wide benthic prey fish population dynamics. Effort increased from 55 bottom trawl sites to 135 trawl sites collected in depths from 8 - 225m (26 – 738 ft). The spatial coverage of sampling was also expanded and occurred in all major lake basins. The resulting distribution of tow depths more closely matched the available lake depth distribution. The additional effort illustrated how previous surveys were underestimating lake-wide Deepwater Sculpin, Myoxocephalus thompsonii, abundance by not sampling in areas of highest density. We also found species richness was greater in the new sampling sites relative to the historic sites with 11 new fish species caught in the new sites including juvenile Round Whitefish, Prosopium cylindraceum, and Mottled sculpin, Cottus bairdii. Species-specific assessments found Slimy Sculpin, Cottus cognatus abundance increased slightly in 2015 relative to 2014, while Deepwater Sculpin and Round Goby, Neogobius melanostomus, dramatically increased in 2015, relative to 2014. The cooperative, lake-wide Benthic Prey Fish Survey expanded our understanding of benthic fish population dynamics and habitat use in Lake Ontario. This survey’s data and interpretations influence international resource management decision making, such as informing the Deepwater Sculpin conservation status and assessing the balance between sport fish consumption and prey fish populations. Additionally a significant Lake Ontario event occurred in May 2015 when a single

Fluorescence in-situ hybridization (FISH) as a tool for visualization and enumeration of Campylobacter in broiler ceca

USDA-ARS?s Scientific Manuscript database

Food-borne human pathogens are typically detected and enumerated by either cultural methods or PCR-based approaches. Fluorescence in-situ hybridization (FISH) is a standard microscopy tool for microbial ecology but has not been widely used for food safety applications despite important advantages o...

Detection of early developmental stages of Myxobolus cerebralis in fish and tubificid oligochaete hosts by in situ hybridization.

PubMed

Antonio, D B; El-Matbouli, M; Hedrick, R P

1999-11-01

The myxosporean and actinosporean spores of Myxobolus cerebralis develop through many stages in their respective hosts, salmonid fishes and a tubificid oligochaete. Using a modified, non-radioactive in situ hybridization protocol, the parasite, which exhibits radically different structural forms during its development in each host, could be specifically detected in paraffin-embedded tissues of both fish and oligochaetes. Our study aims to demonstrate the application of the technique for detection of early stages of M. cerebralis in both hosts.

Methods for assessing fish populations

Treesearch

Kevin L. Pope; Steve E. Lochmann; Michael K. Young

2010-01-01

Fisheries managers are likely to assess fish populations at some point during the fisheries management process. Managers that follow the fisheries management process (see Chapter 5) might find their knowledge base insufficient during the steps of problem identification or management action and must assess a population before appropriate actions can be taken. Managers...

Fish stomach contents in benthic macroinvertebrate assemblage assessments.

PubMed

Tupinambás, T H; Pompeu, P S; Gandini, C V; Hughes, R M; Callisto, M

2015-01-01

The choice of sampling gears to assess benthic macroinvertebrate communities depends on environmental characteristics, study objectives, and cost effectiveness. Because of the high foraging capacity and diverse habitats and behaviors of benthophagous fishes, their stomach contents may offer a useful sampling tool in studies of benthic macroinvertebrates, especially in large, deep, fast rivers that are difficult to sample with traditional sediment sampling gear. Our objective was to compare the benthic macroinvertebrate communities sampled from sediments with those sampled from fish stomachs. We collected benthic macroinvertebrates and fish from three different habitat types (backwater, beach, riffle) in the wet season, drying season, and dry season along a single reach of the Grande River (Paraná River Basin, southeast Brazil). We sampled sediments through use of a Petersen dredge (total of 216 grabs) and used gill nets to sample fish (total of 36 samples). We analyzed the stomach contents of three commonly occurring benthophagous fish species (Eigenmannia virescens, Iheringichthys labrosus, Leporinus amblyrhynchus). Chironomids dominated in both sampling methods. Macroinvertebrate taxonomic composition and abundances from fish stomachs differed from those from sediment samples, but less so from riffles than from backwater and beach habitats. Macroinvertebrate taxa from E. virescens stomachs were more strongly correlated with sediment samples from all three habitats than were those from the other two species. The species accumulation curves and higher mean dispersion values, compared with with sediment samples suggest that E. virescens is more efficient than sediment samples and the other fish studied at collecting benthic taxa. We conclude that by analyzing the stomach contents of benthophagous fishes it is possible to assess important characteristics of benthic communities (dispersion, taxonomic composition and diversity). This is especially true for studies

Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera

NASA Astrophysics Data System (ADS)

Zhang, Qing-Chun; Liu, Qing; Kang, Zhen-Jun; Yu, Ren-Cheng; Yan, Tian; Zhou, Ming-Jiang

2015-09-01

Large-scale green tides have occurred consecutively since 2007 in the Yellow Sea (YS), China. The dominant causative species of the green tides has been identified as Ulva prolifera. The origin of green tides in the YS has been traced back to the Subei Shoal based on the results of remote-sensing, numerical simulations and field investigations. However, it is difficult to study the early development of green tides in the Subei Shoal because of the mixture of multiple green algae and the morphological diversity of U. prolifera when under variable environmental conditions. In this study, a rapid and accurate fluorescence in situ hybridization (FISH) method was developed to detect U. prolifera from the community of green algae targeting the 5S rDNA spacer region of U. prolifera. Two specific probes, 5S-1 and 5S-2, were designed based on the sequences of the 5S rDNA spacer regions of U. prolifera, Ulva linza and Ulva flexuosa. Specificity of the FISH method was tested using the six species of green algae commonly occurring in the Subei Shoal, including U. prolifera, U. linza, U. flexuosa, Ulva compressa, Ulva pertusa and Blidingia sp. The results showed that only U. prolifera could be labeled with both probes. Probe 5S-1, which showed a much higher labeling efficiency on U. prolifera, was ultimately selected as the probe for the FISH detection. The sample preparation method was optimized, particularly for the mature green algae, by the addition of cellulase and proteinase K in the pre-hybridization solution. Labeling efficiency with the probe 5S-1 reached 96% on average under the optimized conditions. The successful development of the FISH method has been applied to qualitative and quantitative analysis of field samples collected from the YS, and the results indicate a potential use in future green algae studies.

Lake Ontario benthic prey fish assessment, 2014

USGS Publications Warehouse

Weidel, Brian C.; Walsh, Maureen

2015-01-01

Benthic prey fishes are an important component of the Lake Ontario fish community and serve as vectors that move energy from benthic invertebrates into native and introduced sport fishes. Since the 1970’s, the USGS Lake Ontario Biological Station has assessed benthic fish populations and community dynamics with bottom trawls at depths ranging from 8 m out to depths of 150-225 m along the south and eastern shores of Lake Ontario. From the late 1970’s through the early 2000’s the benthic fish community was dominated by Slimy Sculpin Cottus cognatus, but in 2004 non-native Round Goby Neogobius melanostomus abundance increased and, since then Round Goby have generally been the dominant benthic species. Over the past 10 years the native Deepwater Sculpin Myoxocephalus thompsonii, once considered absent from the lake, have increased. Presently their lake-wide biomass density is equal to, or larger than, Slimy Sculpin. Species-specific assessments found Slimy and Deepwater Sculpin abundance increased slightly in 2014 relative to 2013, while changes in Round Goby abundance differed between spring and fall survey. Recent survey modifications have increased our understanding of benthic prey fish abundance and behavior in Lake Ontario. For instance, increasing the maximum tow depth to 225 m in 2014 improved our understanding of Deepwater Sculpin distribution in this rarely sampled lake habitat.

Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells

PubMed Central

Barakat, Tahsin Stefan; Gribnau, Joost

2014-01-01

Fluorescent in situ hybridization (FISH) is a molecular technique which enables the detection of nucleic acids in cells. DNA FISH is often used in cytogenetics and cancer diagnostics, and can detect aberrations of the genome, which often has important clinical implications. RNA FISH can be used to detect RNA molecules in cells and has provided important insights in regulation of gene expression. Combining DNA and RNA FISH within the same cell is technically challenging, as conditions suitable for DNA FISH might be too harsh for fragile, single stranded RNA molecules. We here present an easily applicable protocol which enables the combined, simultaneous detection of Xist RNA and DNA encoded by the X chromosomes. This combined DNA-RNA FISH protocol can likely be applied to other systems where both RNA and DNA need to be detected. PMID:24961515

HER2 Gene Amplification Testing by Fluorescent In Situ Hybridization (FISH): Comparison of the ASCO-College of American Pathologists Guidelines With FISH Scores Used for Enrollment in Breast Cancer International Research Group Clinical Trials

PubMed Central

Sauter, Guido; Buyse, Marc; Fourmanoir, Hélène; Quinaux, Emmanuel; Tsao-Wei, Denice D.; Eiermann, Wolfgang; Robert, Nicholas; Pienkowski, Tadeusz; Crown, John; Martin, Miguel; Valero, Vicente; Mackey, John R.; Bee, Valerie; Ma, Yanling; Villalobos, Ivonne; Campeau, Anaamika; Mirlacher, Martina; Lindsay, Mary-Ann; Slamon, Dennis J.

2016-01-01

Purpose ASCO and the College of American Pathologists (ASCO-CAP) recently recommended further changes to the evaluation of human epidermal growth factor receptor 2 gene (HER2) amplification by fluorescent in situ hybridization (FISH). We retrospectively assessed the impact of these new guidelines by using annotated Breast Cancer International Research Group (BCIRG) -005, BCIRG-006, and BCIRG-007 clinical trials data for which we have detailed outcomes. Patients and Methods The HER2 FISH status of BCIRG-005/006/007 patients with breast cancers was re-evaluated according to current ASCO-CAP guidelines, which designates five different groups according to HER2 FISH ratio and average HER2 gene copy number per tumor cell: group 1 (in situ hybridization [ISH]–positive): HER2-to-chromosome 17 centromere ratio ≥ 2.0, average HER2 copies ≥ 4.0; group 2 (ISH-positive): ratio ≥ 2.0, copies < 4.0; group 3 (ISH-positive): ratio < 2.0, copies ≥ 6.0; group 4 (ISH-equivocal): ratio < 2.0, copies ≥ 4.0 and < 6.0; and group 5 (ISH-negative): ratio < 2.0, copies < 4.0. We assessed correlations with HER2 protein, clinical outcomes by disease-free survival (DFS) and overall survival (OS) and benefit from trastuzumab therapy (hazard ratio [HR]). Results Among 10,468 patients with breast cancers who were successfully screened for trial entry, 40.8% were in ASCO-CAP ISH group 1, 0.7% in group 2; 0.5% in group 3, 4.1% in group 4, and 53.9% in group 5. Distributions were similar in screened compared with accrued subpopulations. Among accrued patients, FISH group 1 breast cancers were strongly correlated with immunohistochemistry 3+ status (P < .0001), whereas groups 2, 3, 4, and 5 were not; however, groups 2, 4 and, 5 were strongly correlated with immunohistochemistry 0/1+ status (all P < .0001), whereas group 3 was not. Among patients accrued to BCIRG-005, group 4 was not associated with significantly worse DFS or OS compared with group 5. Among patients accrued to BCIRG-006

HER2 Gene Amplification Testing by Fluorescent In Situ Hybridization (FISH): Comparison of the ASCO-College of American Pathologists Guidelines With FISH Scores Used for Enrollment in Breast Cancer International Research Group Clinical Trials.

PubMed

Press, Michael F; Sauter, Guido; Buyse, Marc; Fourmanoir, Hélène; Quinaux, Emmanuel; Tsao-Wei, Denice D; Eiermann, Wolfgang; Robert, Nicholas; Pienkowski, Tadeusz; Crown, John; Martin, Miguel; Valero, Vicente; Mackey, John R; Bee, Valerie; Ma, Yanling; Villalobos, Ivonne; Campeau, Anaamika; Mirlacher, Martina; Lindsay, Mary-Ann; Slamon, Dennis J

2016-10-10

Purpose ASCO and the College of American Pathologists (ASCO-CAP) recently recommended further changes to the evaluation of human epidermal growth factor receptor 2 gene (HER2) amplification by fluorescent in situ hybridization (FISH). We retrospectively assessed the impact of these new guidelines by using annotated Breast Cancer International Research Group (BCIRG) -005, BCIRG-006, and BCIRG-007 clinical trials data for which we have detailed outcomes. Patients and Methods The HER2 FISH status of BCIRG-005/006/007 patients with breast cancers was re-evaluated according to current ASCO-CAP guidelines, which designates five different groups according to HER2 FISH ratio and average HER2 gene copy number per tumor cell: group 1 (in situ hybridization [ISH]-positive): HER2-to-chromosome 17 centromere ratio ≥ 2.0, average HER2 copies ≥ 4.0; group 2 (ISH-positive): ratio ≥ 2.0, copies < 4.0; group 3 (ISH-positive): ratio < 2.0, copies ≥ 6.0; group 4 (ISH-equivocal): ratio < 2.0, copies ≥ 4.0 and < 6.0; and group 5 (ISH-negative): ratio < 2.0, copies < 4.0. We assessed correlations with HER2 protein, clinical outcomes by disease-free survival (DFS) and overall survival (OS) and benefit from trastuzumab therapy (hazard ratio [HR]). Results Among 10,468 patients with breast cancers who were successfully screened for trial entry, 40.8% were in ASCO-CAP ISH group 1, 0.7% in group 2; 0.5% in group 3, 4.1% in group 4, and 53.9% in group 5. Distributions were similar in screened compared with accrued subpopulations. Among accrued patients, FISH group 1 breast cancers were strongly correlated with immunohistochemistry 3+ status (P < .0001), whereas groups 2, 3, 4, and 5 were not; however, groups 2, 4 and, 5 were strongly correlated with immunohistochemistry 0/1+ status (all P < .0001), whereas group 3 was not. Among patients accrued to BCIRG-005, group 4 was not associated with significantly worse DFS or OS compared with group 5. Among patients accrued to BCIRG-006, only

Trade-off between morphological convergence and opportunistic diet behavior in fish hybrid zone

PubMed Central

Corse, Emmanuel; Costedoat, Caroline; Pech, Nicolas; Chappaz, Rémi; Grey, Jonathan; Gilles, André

2009-01-01

Background The invasive Chondrostoma nasus nasus has colonized part of the distribution area of the protected endemic species Chondrostoma toxostoma toxostoma. This hybrid zone is a complex system where multiple effects such as inter-species competition, bi-directional introgression, strong environmental pressure and so on are combined. Why do sympatric Chondrostoma fish present a unidirectional change in body shape? Is this the result of inter-species interactions and/or a response to environmental effects or the result of trade-offs? Studies focusing on the understanding of a trade-off between multiple parameters are still rare. Although this has previously been done for Cichlid species flock and for Darwin finches, where mouth or beak morphology were coupled to diet and genetic identification, no similar studies have been done for a fish hybrid zone in a river. We tested the correlation between morphology (body and mouth morphology), diet (stable carbon and nitrogen isotopes) and genomic combinations in different allopatric and sympatric populations for a global data set of 1330 specimens. To separate the species interaction effect from the environmental effect in sympatry, we distinguished two data sets: the first one was obtained from a highly regulated part of the river and the second was obtained from specimens coming from the less regulated part. Results The distribution of the hybrid combinations was different in the two part of the sympatric zone, whereas all the specimens presented similar overall changes in body shape and in mouth morphology. Sympatric specimens were also characterized by a larger diet behavior variance than reference populations, characteristic of an opportunistic diet. No correlation was established between the body shape (or mouth deformation) and the stable isotope signature. Conclusion The Durance River is an untamed Mediterranean river despite the presence of numerous dams that split the river from upstream to downstream. The

Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

DOE PAGES

Cui, Yi; Hu, Dehong; Markillie, Lye Meng; ...

2017-10-04

Here, quantitative gene expression analysis in intact single cells can be achieved using single molecule-based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple oligonucleotide probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific probe binding and tissue autofluorescence, especially when only a small number of probes can be fitted to the target transcript. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on on-off duty cycles of photoswitchable dyes.more » This fluctuation localization imaging-based FISH (fliFISH) uses on-time fractions (measured over a series of exposures) collected from transcripts bound to as low as 8 probes, which are distinct from on-time fractions collected from nonspecifically bound probes or autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2 and their ratio ( Nkx2- 2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct on-time fractions, laying the foundation for reliable single-cell transcriptomics.« less

Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cui, Yi; Hu, Dehong; Markillie, Lye Meng

Here, quantitative gene expression analysis in intact single cells can be achieved using single molecule-based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple oligonucleotide probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific probe binding and tissue autofluorescence, especially when only a small number of probes can be fitted to the target transcript. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on on-off duty cycles of photoswitchable dyes.more » This fluctuation localization imaging-based FISH (fliFISH) uses on-time fractions (measured over a series of exposures) collected from transcripts bound to as low as 8 probes, which are distinct from on-time fractions collected from nonspecifically bound probes or autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2 and their ratio ( Nkx2- 2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct on-time fractions, laying the foundation for reliable single-cell transcriptomics.« less

Concurrent assessment of fish and habitat in warmwater streams in Wyoming

USGS Publications Warehouse

Quist, M.C.; Hubert, W.A.; Rahel, F.J.

2006-01-01

Fisheries research and management in North America have focused largely on sport fishes, but native non-game fishes have attracted increased attention due to their declines. The Warmwater Stream Assessment (WSA) was developed to evaluate simultaneously both fish and habitat in Wyoming streams by a process that includes three major components: (1) stream-reach selection and accumulation of existing information, (2) fish and habitat sampling and (3) summarisation and evaluation of fish and habitat information. Fish are sampled by electric fishing or seining and habitat is measured at reach and channel-unit (i.e. pool, run, riffle, side channel, or backwater) scales. Fish and habitat data are subsequently summarised using a data-matrix approach. Hierarchical decision trees are used to assess critical habitat requirements for each fish species expected or found in the reach. Combined measurements of available habitat and the ecology of individual species contribute to the evaluation of the observed fish assemblage. The WSA incorporates knowledge of the fish assemblage and habitat features to enable inferences of factors likely influencing both the fish assemblage and their habitat. The WSA was developed for warmwater streams in Wyoming, but its philosophy, process and conceptual basis may be applied to environmental assessments in other geographical areas. ?? 2006 Blackwell Publishing Ltd.

Assessing consumer awareness about mercury in fish.

PubMed

Ratnapradipa, Dhitinut; Quilliam, Daniela; Wier, Lauren M; Migliore, Beverly; Dundulis, William

2009-12-01

The study discussed in this article was conducted to determine if a retail-based educational campaign would be an effective tool to inform consumers about mercury in fish. In 2005, the Rhode Island (RI) Department of Health, in conjunction with the RI Food Dealers Association, conducted surveys in eight supermarkets to assess consumers' knowledge and awareness of mercury in fish. A presurvey was administered to 523 respondents as they prepared to exit the store June 17-19. During the week of July 11-17, a "Fish Week" educational program was conducted. An identical postsurvey to evaluate the effectiveness of "Fish Week" was administered to 556 customers exiting participating supermarkets on July 21-24. A significant (p < .05) increase occurred in the number of respondents who identified brochures as an information source about mercury in fish. Small, but significant, decreases in awareness and knowledge about mercury in fish issues, however, were unexpected findings that were likely due to factors discussed.

FISH-Flow, a protocol for the concurrent detection of mRNA and protein in single cells using fluorescence in situ hybridization and flow cytometry

PubMed Central

Arrigucci, Riccardo; Bushkin, Yuri; Radford, Felix; Lakehal, Karim; Vir, Pooja; Pine, Richard; Martin, December; Sugarman, Jeffrey; Zhao, Yanlin; Yap, George S; Lardizabal, Alfred A; Tyagi, Sanjay; Gennaro, Maria Laura

2017-01-01

We describe a flow-cytometry-based protocol for intracellular mRNA measurements in nonadherent mammalian cells using fluorescence in situ hybridization (FISH) probes. The method, which we call FISH-Flow, allows for high-throughput multiparametric measurements of gene expression, a task that was not feasible with earlier, microscopy-based approaches. The FISH-Flow protocol involves cell fixation, permeabilization and hybridization with a set of fluorescently labeled oligonucleotide probes. In this protocol, surface and intracellular protein markers can also be stained with fluorescently labeled antibodies for simultaneous protein and mRNA measurement. Moreover, a semiautomated, single-tube version of the protocol can be performed with a commercially available cell-wash device that reduces cell loss, operator time and interoperator variability. It takes ~30 h to perform this protocol. An example of FISH-Flow measurements of cytokine mRNA induction by ex vivo stimulation of primed T cells with specific antigens is described. PMID:28518171

Increased incidence of micronuclei assessed with the micronucleus assay and the fluorescence in situ hybridization (FISH) technique in peripheral blood lymphocytes of nurses exposed to nitrous oxide.

PubMed

Lewińska, D; Stepnik, M; Krajewski, W; Arkusz, J; Stańczyk, M; Wrońska-Nofer, T

2005-03-07

It has been postulated that exposure to nitrous oxide and halogenated anaesthetics is associated with various adverse health effects such as neurological and reproductive abnormalities or impairment of hepatic functions. In spite of the quite well known genotoxic effects of exposure to nitrous oxide in vivo, the mechanisms of these effects are still not clear. The aim of this study was to assess the frequency of micronuclei and to identify the type of chromosomal damage (clastogenic or aneugenic) in peripheral blood lymphocytes of operating-room nurses exposed to nitrous oxide. The study group comprised 46 women working at departments where the concentration of nitrous oxide ranged from 14 to 2308 mg/m3. The control population was composed of 28 women employed in the same hospitals but in non-surgical departments. The clastogenic/aneugenic effect of nitrous oxide was evaluated in lymphocytes using the standard micronucleus (MN) assay in combination with the fluorescence in situ hybridization (FISH) technique with pancentromeric probes. The results show a significant increase of the MN frequency in lymphocytes of exposed nurses compared with the control group (4.36+/-2.23 versus 9.02+/-4.67). The multiple regression analysis revealed a statistically significant relationship (p=0.0009) between MN frequency and exposure status, indicating that the level of exposure was the main factor affecting chromosomal damage. As assessed by FISH analysis, the overall frequencies of centromere-positive MN in the control and exposed groups were 43 and 49%, respectively. The increase observed in the exposed group may suggest a slight, statistically insignificant pro-aneugenic effect of exposure to nitrous oxide.

Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice

PubMed Central

Pathak, Rupak; Koturbash, Igor; Hauer-Jensen, Martin

2017-01-01

Ionizing radiation (IR) induces numerous stable and unstable chromosomal aberrations. Unstable aberrations, where chromosome morphology is substantially compromised, can easily be identified by conventional chromosome staining techniques. However, detection of stable aberrations, which involve exchange or translocation of genetic materials without considerable modification in the chromosome morphology, requires sophisticated chromosome painting techniques that rely on in situ hybridization of fluorescently labeled DNA probes, a chromosome painting technique popularly known as fluorescence in situ hybridization (FISH). FISH probes can be specific for whole chromosome/s or precise sub-region on chromosome/s. The method not only allows visualization of stable aberrations, but it can also allow detection of the chromosome/s or specific DNA sequence/s involved in a particular aberration formation. A variety of chromosome painting techniques are available in cytogenetics; here two highly sensitive methods, multiple fluorescence in situ hybridization (mFISH) and spectral karyotyping (SKY), are discussed to identify inter-chromosomal stable aberrations that form in the bone marrow cells of mice after exposure to total body irradiation. Although both techniques rely on fluorescent labeled DNA probes, the method of detection and the process of image acquisition of the fluorescent signals are different. These two techniques have been used in various research areas, such as radiation biology, cancer cytogenetics, retrospective radiation biodosimetry, clinical cytogenetics, evolutionary cytogenetics, and comparative cytogenetics. PMID:28117817

A phylogeographic investigation of the hybrid origin of a species of swordtail fish from Mexico.

PubMed

Jones, Julia C; Perez-Sato, Juan-Antonio; Meyer, Axel

2012-06-01

Hybrid speciation may contribute significantly to generating biodiversity, but only a few well-documented examples for it exist so far that do not involve polyploidization as a mechanism. The swordtail fish, Xiphophorus clemenciae, shows common hallmarks of a hybrid origin and still overlaps in its current geographic distribution with its putative ancestral species (Xiphophorus hellerii and Xiphophorus maculatus). Xiphophorus clemenciae provides an ideal system for investigating the possible continued genetic interactions between a hybrid and its parental species. Here, we use microsatellite and mitochondrial markers to investigate the population structure of these species of swordtails and search for signs of recent hybridization. Individuals were sampled from 21 localities across the known range of X. clemenciae- the Isthmus of Tehuantepec (IT) Mexico, and several environmental parameters that might represent barriers to dispersal were recorded. The hybridization event that gave rise to X. clemenciae appears to be rather ancient, and a single origin is likely. We find negligible evidence for ongoing hybridization and introgression between the putative ancestral species, because they now occupy distinct ecological niches, and a common haplotype is shared by most populations of X. clemenciae. The population structure within these species shows an isolation-by-distance (IBD) pattern and genetic differentiation between most populations is significant and high. We infer that tectonic evolution in the Isthmus has greatly restricted gene flow between the southern and central IT populations of X. clemenciae and X. helleriii and provide preliminary information to aid in conservation management of this geographically restricted hybrid species, X. clemenciae. © 2012 Blackwell Publishing Ltd.

Fluorescence in situ hybridization (CARD-FISH) of microorganisms in hydrocarbon contaminated aquifer sediment samples.

PubMed

Tischer, Karolin; Zeder, Michael; Klug, Rebecca; Pernthaler, Jakob; Schattenhofer, Martha; Harms, Hauke; Wendeberg, Annelie

2012-12-01

Groundwater ecosystems are the most important sources of drinking water worldwide but they are threatened by contamination and overexploitation. Petroleum spills account for the most common source of contamination and the high carbon load results in anoxia and steep geochemical gradients. Microbes play a major role in the transformation of petroleum hydrocarbons into less toxic substances. To investigate microbial populations at the single cell level, fluorescence in situ hybridization (FISH) is now a well-established technique. Recently, however, catalyzed reporter deposition (CARD)-FISH has been introduced for the detection of microbes from oligotrophic environments. Nevertheless, petroleum contaminated aquifers present a worst case scenario for FISH techniques due to the combination of high background fluorescence of hydrocarbons and the presence of small microbial cells caused by the low turnover rates characteristic of groundwater ecosystems. It is therefore not surprising that studies of microorganisms from such sites are mostly based on cultivation techniques, fingerprinting, and amplicon sequencing. However, to reveal the population dynamics and interspecies relationships of the key participants of contaminant degradation, FISH is an indispensable tool. In this study, a protocol for FISH was developed in combination with cell quantification using an automated counting microscope. The protocol includes the separation and purification of microbial cells from sediment particles, cell permeabilization and, finally, CARD-FISH in a microwave oven. As a proof of principle, the distribution of Archaea and Bacteria was shown in 60 sediment samples taken across the contaminant plume of an aquifer (Leuna, Germany), which has been heavily contaminated with several ten-thousand tonnes of petroleum hydrocarbons since World War II. Copyright © 2012 Elsevier GmbH. All rights reserved.

A UK NEQAS ICC and ISH multicentre study using the Kreatech Poseidon HER2 FISH probe: intersite variation can be rigorously controlled using FISH.

PubMed

Bartlett, John M S; Campbell, Fiona M; Ibrahim, Merdol; Thomas, Jeremy; Wencyk, Pete; Ellis, Ian; Kay, Elaine; Connolly, Yvonne; O'Grady, Anthony; Barnett, Sarah; Starczynski, Jane; Cunningham, Paul; Miller, Keith

2010-02-01

To assess a new HER2 fluorescence in situ hybridization (FISH) test and report on multicentre intrasite and intersite variation. HER2 results were scored from 45 breast cancers in eight laboratories using the Kreatech Poseidon HER2 FISH probe (Kreatech Diagnostics, Amsterdam, the Netherlands). Overall, 80.9% of cores were successfully analysed. Mean intrasite variation for HER2 ratio assessment was low (4.74%). Intersite variation in ratio was in line with previous reports (11.9+/-0.8%) for both reference and non-reference laboratories; only one laboratory displayed significantly higher intersite variation (P=0.009) than the remaining seven laboratories. The overall incidence of misclassification of cores was <1.3%, demonstrating an excellent level of concordance (>98.7%) across all eight laboratories, irrespective of whether they were 'reference' or 'routine diagnostic' laboratories. The Kreatech Poseidon HER2 FISH test is robust and reproducible. Highly quantitatively reproducible FISH results were obtained from eight 'diagnostic' and 'reference' laboratories; however, continued quality assessments are essential to good performance.

Abnormalities of hybrid grouper (Epinephelus fuscoguttatus x Epinephelus lanceolatus) in Situbondo

NASA Astrophysics Data System (ADS)

Triastuti, J.; Pursetyo, K. T.; Monica, A.; Lutfiyah, L.; Budi, D. S.

2018-04-01

Grouper is one of consumption fish which is demanded excessively by local consumers and foreign consumers. Hybridization of grouper has been performed considerably that produce the good genetic quality of hybrid variants. One of grouper fish which has good genetic in its growth is kertang grouper fish. Nowadays many hatcheries performing hybridization between kertang grouper fish and tiger grouper fish, however observation of the hybrid abnormality has not been performed yet. Abnormality is able to increase since genetic causes, so that observation of abnormality occurrence in cantang hybrid grouper fish in Situbondo, JawaTimur, Indonesia in May – July in 3 times grading of juvenile stadia was performed. Results showed abnormalities were observed on mouth and operculum, branched of neural arch, fusion of neural arch, fusion of posterior truncus vertebrae, fusion of caudal vertebrae, fusion of anterior truncus vertebrae.

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

PubMed Central

Ronander, Elena; Bengtsson, Dominique C.; Joergensen, Louise; Jensen, Anja T. R.; Arnot, David E.

2012-01-01

Adhesion of Plasmodium falciparum infected erythrocytes (IE) to human endothelial receptors during malaria infections is mediated by expression of PfEMP1 protein variants encoded by the var genes. The haploid P. falciparum genome harbors approximately 60 different var genes of which only one has been believed to be transcribed per cell at a time during the blood stage of the infection. How such mutually exclusive regulation of var gene transcription is achieved is unclear, as is the identification of individual var genes or sub-groups of var genes associated with different receptors and the consequence of differential binding on the clinical outcome of P. falciparum infections. Recently, the mutually exclusive transcription paradigm has been called into doubt by transcription assays based on individual P. falciparum transcript identification in single infected erythrocytic cells using RNA fluorescent in situ hybridization (FISH) analysis of var gene transcription by the parasite in individual nuclei of P. falciparum IE1. Here, we present a detailed protocol for carrying out the RNA-FISH methodology for analysis of var gene transcription in single-nuclei of P. falciparum infected human erythrocytes. The method is based on the use of digoxigenin- and biotin- labeled antisense RNA probes using the TSA Plus Fluorescence Palette System2 (Perkin Elmer), microscopic analyses and freshly selected P. falciparum IE. The in situ hybridization method can be used to monitor transcription and regulation of a variety of genes expressed during the different stages of the P. falciparum life cycle and is adaptable to other malaria parasite species and other organisms and cell types. PMID:23070076

Responses to crizotinib in patients with ALK-positive lung adenocarcinoma who tested immunohistochemistry (IHC)-positive and fluorescence in situ hybridization (FISH)-negative.

PubMed

Ma, Di; Wang, Zheng; Yang, Lin; Mu, Xinlin; Wang, Yan; Zhao, Xinming; Li, Junling; Lin, Dongmei

2016-09-27

Although the Ventana immunohistochemistry (IHC) platform for detecting anaplastic lymphoma kinase gene (ALK) (D5F3) expression was recently approved by the US Food and Drugs Administration (FDA), fluorescence in situ hybridization (FISH) is still the "gold-standard" method recommended by the US National Comprehensive Cancer Network (NCCN) guideline for NSCLC. We evaluated 6 ALK-positive lung adenocarcinoma patients who tested Ventana IHC-positive and FISH-negative and assessed their clinical responses to the ALK tyrosine kinase inhibitor (TKI) crizotinib. Histologic and cytologic specimens from the 6 patients were stained with Ventana anti-ALK(D5F3) rabbit monoclonal primary antibody using the OptiView™ DAB IHC detection kit and OptiView™ amplification kit on a Ventana BenchMark XT processor. In addition, they were also tested by FISH, qRT-PCR, next-generation sequencing (NGS), and RNAscope ISH analysis. All patients received crizotinib treatment and their follow-up clinical data were recorded. The objective response rate achieved with crizotinib therapy was 66.7% (4/6 partial responses and 2/6 stable disease). One patient in whom a new fusion type (EML4->EXOC6B->ALK fusion) was identified obtained a partial response. These findings indicate that patients with ALK-positive lung adenocarcinoma who test Ventana IHC-positive and FISH-negative may still respond to crizotinib therapy.

Effectiveness of fishing gears to assess fish assemblage size structure in small lake ecosystems

Treesearch

T. A. Clement; K. Pangle; D. G. Uzarski; B. A. Murry

2014-01-01

Measurement of fish body-size distributions is increasingly used as a management tool to assess fishery status. However, the effects of gear selection on observed fish size structure has not received sufficient attention. Four different gear types (experimental gill nets, fine mesh bag seine, and two different sized mesh trap nets), which are commonly employed in the...

Utility and impact of early t(15;17) identification by Fluorescence In Situ Hybridization (FISH) in clinical decision making for patients in Acute Promyelocytic Leukemia (APL).

PubMed

Kolhe, R; Mangaonkar, A; Mansour, J; Clemmons, A; Shaw, J; Dupont, B; Walczak, L; Mondal, A; Rojiani, A; Jillella, A; Kota, V

2015-08-01

Acute Promyelocytic Leukemia (APL) is a curable malignancy with studies showing above 90% survival. However, population-based studies looking at survival suggest that approximately 30% of patients with APL die during induction. Early demonstration of t(15;17) will lead to accurate decision making regarding treatment. The aim of this project was to validate earlier time frames for the Abbott Molecular Vysis LSI promyelocytic leukemia (PML)/ retinoic acid receptor alpha (RARA) fluorescence in situ hybridization (FISH) probe (ASR 6-16 h). Twenty patients (15 APL cases and five non-APL cases) were selected for validating various hybridization times for the FISH probe. Expected normal signal pattern was two red and two green signals (2R2G), and the most common expected abnormal signal pattern was two fusion (yellow) signals, one red and one green (2F1R1G) and/or one fusion, one red and one green (1F1R1G). The specificity of the probe ranged from 84% at 2 h, 86% at 4 h, 84% at 6 h, and 87% for overnight hybridization. The sensitivity increased from 79% at 2 h, 80% at 4 h, 81% at 6 h to 87% for overnight hybridization. Based on the validation studies, we recommend reading of FISH results at the 4-h incubation mark for a preliminary diagnosis and confirmation with overnight hybridization. © 2015 John Wiley & Sons Ltd.

A moving target—incorporating knowledge of the spatial ecology of fish into the assessment and management of freshwater fish populations

USGS Publications Warehouse

Cooke, Steven J.; Martins, Eduardo G; Struthers, Daniel P; Gutowsky, Lee F G; Powers, Michael H.; Doka, Susan E.; Dettmers, John M.; Crook, David A; Lucas, Martyn C.; Holbrook, Christopher; Krueger, Charles C.

2016-01-01

Freshwater fish move vertically and horizontally through the aquatic landscape for a variety of reasons, such as to find and exploit patchy resources or to locate essential habitats (e.g., for spawning). Inherent challenges exist with the assessment of fish populations because they are moving targets. We submit that quantifying and describing the spatial ecology of fish and their habitat is an important component of freshwater fishery assessment and management. With a growing number of tools available for studying the spatial ecology of fishes (e.g., telemetry, population genetics, hydroacoustics, otolith microchemistry, stable isotope analysis), new knowledge can now be generated and incorporated into biological assessment and fishery management. For example, knowing when, where, and how to deploy assessment gears is essential to inform, refine, or calibrate assessment protocols. Such information is also useful for quantifying or avoiding bycatch of imperiled species. Knowledge of habitat connectivity and usage can identify critically important migration corridors and habitats and can be used to improve our understanding of variables that influence spatial structuring of fish populations. Similarly, demographic processes are partly driven by the behavior of fish and mediated by environmental drivers. Information on these processes is critical to the development and application of realistic population dynamics models. Collectively, biological assessment, when informed by knowledge of spatial ecology, can provide managers with the ability to understand how and when fish and their habitats may be exposed to different threats. Naturally, this knowledge helps to better evaluate or develop strategies to protect the long-term viability of fishery production. Failure to understand the spatial ecology of fishes and to incorporate spatiotemporal data can bias population assessments and forecasts and potentially lead to ineffective or counterproductive management actions.

A moving target--incorporating knowledge of the spatial ecology of fish into the assessment and management of freshwater fish populations.

PubMed

Cooke, Steven J; Martins, Eduardo G; Struthers, Daniel P; Gutowsky, Lee F G; Power, Michael; Doka, Susan E; Dettmers, John M; Crook, David A; Lucas, Martyn C; Holbrook, Christopher M; Krueger, Charles C

2016-04-01

Freshwater fish move vertically and horizontally through the aquatic landscape for a variety of reasons, such as to find and exploit patchy resources or to locate essential habitats (e.g., for spawning). Inherent challenges exist with the assessment of fish populations because they are moving targets. We submit that quantifying and describing the spatial ecology of fish and their habitat is an important component of freshwater fishery assessment and management. With a growing number of tools available for studying the spatial ecology of fishes (e.g., telemetry, population genetics, hydroacoustics, otolith microchemistry, stable isotope analysis), new knowledge can now be generated and incorporated into biological assessment and fishery management. For example, knowing when, where, and how to deploy assessment gears is essential to inform, refine, or calibrate assessment protocols. Such information is also useful for quantifying or avoiding bycatch of imperiled species. Knowledge of habitat connectivity and usage can identify critically important migration corridors and habitats and can be used to improve our understanding of variables that influence spatial structuring of fish populations. Similarly, demographic processes are partly driven by the behavior of fish and mediated by environmental drivers. Information on these processes is critical to the development and application of realistic population dynamics models. Collectively, biological assessment, when informed by knowledge of spatial ecology, can provide managers with the ability to understand how and when fish and their habitats may be exposed to different threats. Naturally, this knowledge helps to better evaluate or develop strategies to protect the long-term viability of fishery production. Failure to understand the spatial ecology of fishes and to incorporate spatiotemporal data can bias population assessments and forecasts and potentially lead to ineffective or counterproductive management actions.

Logging while fishing: An alternate method to cut and thread fishing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tollefsen, E.; Crary, S.; Flores, B.

1996-12-31

New technology has been introduced to allow completion of the wireline logging program after the tool string has become lodged in the wellbore. Charges associated with extracting a stuck tool are substantial. These charges result from the nonproductive time during the fishing trip, an associated wiper trip, and re-logging the well. The ability to continue the logging program while retrieving the logging string from the wellbore is needed. Logging While Fishing (LWF) is a hybrid of existing technologies combined with a new sub capable of severing a cable remotely. This new method is comprised of cut and thread fishing, drillpipemore » conveyed logging, and bridled tool techniques. Utilizing these techniques it is possible to complete wireline logging operations while removing a stuck tool from the wellbore. Completing logging operations using this hybrid method will save operating companies time and money. Other benefits, depending on the situation, include reduced fishing time and an increased level of safety. This application has been demonstrated on jobs in the Gulf of Mexico, North Sea, Venezuela, and Southeast Asia.« less

Analysis of HER2 status in breast carcinoma by fully automated HER2 fluorescence in situ hybridization (FISH): comparison of two immunohistochemical tests and manual FISH.

PubMed

Yoon, Nara; Do, In-Gu; Cho, Eun Yoon

2014-09-01

Easy and accurate HER2 testing is essential when considering the prognostic and predictive significance of HER2 in breast cancer. The use of a fully automated, quantitative FISH assay would be helpful to detect HER2 amplification in breast cancer tissue specimens with reduced inter-laboratory variability. We compared the concordance of HER2 status as assessed by an automated FISH staining system to manual FISH testing. Using 60 formalin-fixed paraffin-embedded breast carcinoma specimens, we assessed HER2 immunoexpression with two antibodies (DAKO HercepTest and CB11). In addition, HER2 status was evaluated with automated FISH using the Leica FISH System for BOND and a manual FISH using the Abbott PathVysion DNA Probe Kit. All but one specimen were successfully stained using both FISH methods. When the data were divided into two groups according to HER2/CEP17 ratio, positive and negative, the results from both the automated and manual FISH techniques were identical for all 59 evaluable specimens. The HER2 and CEP17 copy numbers and HER2/CEP17 ratio showed great agreement between both FISH methods. The automated FISH technique was interpretable with signal intensity similar to those of the manual FISH technique. In contrast with manual FISH, the automated FISH technique showed well-preserved architecture due to low membrane digestion. HER2 immunohistochemistry and FISH results showed substantial significant agreement (κ = 1.0, p < 0.001). HER2 status can be reliably determined using a fully automated HER2 FISH system with high concordance to the well-established manual FISH method. Because of stable signal intensity and high staining quality, the automated FISH technique may be more appropriate than manual FISH for routine applications. © 2013 APMIS. Published by John Wiley & Sons Ltd.

Reagent and labor cost optimization through automation of fluorescence in situ hybridization (FISH) with the VP 2000: an Italian case study.

PubMed

Zanatta, Lucia; Valori, Laura; Cappelletto, Eleonora; Pozzebon, Maria Elena; Pavan, Elisabetta; Dei Tos, Angelo Paolo; Merkle, Dennis

2015-02-01

In the modern molecular diagnostic laboratory, cost considerations are of paramount importance. Automation of complex molecular assays not only allows a laboratory to accommodate higher test volumes and throughput but also has a considerable impact on the cost of testing from the perspective of reagent costs, as well as hands-on time for skilled laboratory personnel. The following study tracked the cost of labor (hands-on time) and reagents for fluorescence in situ hybridization (FISH) testing in a routine, high-volume pathology and cytogenetics laboratory in Treviso, Italy, over a 2-y period (2011-2013). The laboratory automated FISH testing with the VP 2000 Processor, a deparaffinization, pretreatment, and special staining instrument produced by Abbott Molecular, and compared hands-on time and reagent costs to manual FISH testing. The results indicated significant cost and time saving when automating FISH with VP 2000 when more than six FISH tests were run per week. At 12 FISH assays per week, an approximate total cost reduction of 55% was observed. When running 46 FISH specimens per week, the cost saving increased to 89% versus manual testing. The results demonstrate that the VP 2000 processor can significantly reduce the cost of FISH testing in diagnostic laboratories. © 2014 Society for Laboratory Automation and Screening.

Enhancing fish Underwater Visual Census to move forward assessment of fish assemblages: An application in three Mediterranean Marine Protected Areas

PubMed Central

Thiriet, Pierre; Di Franco, Antonio; Francour, Patrice

2017-01-01

Monitoring fish assemblages is needed to assess whether Marine Protected Areas (MPAs) are meeting their conservation and fisheries management goals, as it allows one to track the progress of recovery of exploited species and associated communities. Underwater Visual Census techniques (UVC) are used to monitor fish assemblages in MPAs. UVCs should be adapted to fish abundance, body-size and behaviour, which can strongly affect fish detectability. In Mediterranean subtidal habitats, however, UVC strip transects of one surface area (25x5 m2) are commonly used to survey the whole fish assemblage, from large shy fish to small crypto-benthic fish. Most high trophic level predators (HTLPs) are large shy fish which rarely swim close to divers and, consequently, their abundance may be under-estimated with commonly used transects. Here, we propose an improvement to traditional transect surveys to better account for differences in behaviour among and within species. First, we compared the effectiveness of combining two transect surface areas (large: 35x20 m2; medium: 25x5 m2) in quantifying large, shy fish within and outside Mediterranean MPAs. We identified species-specific body-size thresholds defining a smaller and a larger size class better sampled by medium and large transects respectively. Combining large and medium transects provided more accurate biomass and species richness estimates for large, shy species than using medium transects alone. We thus combined the new approach with two other transect surface areas commonly used to survey crypto-benthic (10x1 m2) and necto-benthic (25x5 m2) species in order to assess how effectively MPAs protection the whole fish assemblage. We verified that MPAs offer significant protection for HTLPs, their response in terms of biomass and density increase in MPAs was always higher in magnitude than other functional groups. Inside MPAs, the contribution of HTLP reached >25% of total fish biomass, against < 2% outside MPAs. Surveys with

Enhancing fish Underwater Visual Census to move forward assessment of fish assemblages: An application in three Mediterranean Marine Protected Areas.

PubMed

Prato, Giulia; Thiriet, Pierre; Di Franco, Antonio; Francour, Patrice

2017-01-01

Monitoring fish assemblages is needed to assess whether Marine Protected Areas (MPAs) are meeting their conservation and fisheries management goals, as it allows one to track the progress of recovery of exploited species and associated communities. Underwater Visual Census techniques (UVC) are used to monitor fish assemblages in MPAs. UVCs should be adapted to fish abundance, body-size and behaviour, which can strongly affect fish detectability. In Mediterranean subtidal habitats, however, UVC strip transects of one surface area (25x5 m2) are commonly used to survey the whole fish assemblage, from large shy fish to small crypto-benthic fish. Most high trophic level predators (HTLPs) are large shy fish which rarely swim close to divers and, consequently, their abundance may be under-estimated with commonly used transects. Here, we propose an improvement to traditional transect surveys to better account for differences in behaviour among and within species. First, we compared the effectiveness of combining two transect surface areas (large: 35x20 m2; medium: 25x5 m2) in quantifying large, shy fish within and outside Mediterranean MPAs. We identified species-specific body-size thresholds defining a smaller and a larger size class better sampled by medium and large transects respectively. Combining large and medium transects provided more accurate biomass and species richness estimates for large, shy species than using medium transects alone. We thus combined the new approach with two other transect surface areas commonly used to survey crypto-benthic (10x1 m2) and necto-benthic (25x5 m2) species in order to assess how effectively MPAs protection the whole fish assemblage. We verified that MPAs offer significant protection for HTLPs, their response in terms of biomass and density increase in MPAs was always higher in magnitude than other functional groups. Inside MPAs, the contribution of HTLP reached >25% of total fish biomass, against < 2% outside MPAs. Surveys with

Benthic prey fish assessment, Lake Ontario 2013

USGS Publications Warehouse

Weidel, Brian C.; Walsh, Maureen; Connerton, Michael J.

2014-01-01

The 2013 benthic fish assessment was delayed and shortened as a result of the U.S. Government shutdown, however the assessment collected 51 of the 62 planned bottom trawls. Over the past 34 years, Slimy Sculpin abundance in Lake Ontario has fluctuated, but ultimately decreased by two orders of magnitude, with a substantial decline occurring in the past 10 years. The 2013 Slimy Sculpin mean bottom trawl catch density (0.001 ind.·m-2, s.d.= 0.0017, n = 52) and mean biomass density (0.015 g·m-2 , s.d.= 0.038, n = 52) were the lowest recorded in the 27 years of sampling using the original bottom trawl design. From 2011-2013, the Slimy Sculpin density and biomass density has decreased by approximately 50% each year. Spring bottom trawl catches illustrate Slimy Sculpin and Round Goby Neogobius melanostoma winter habitat overlaps for as much as 7 months out of a year, providing opportunities for competition and predation. Invasive species, salmonid piscivory, and declines in native benthic invertebrates are likely all important drivers of Slimy Sculpin population dynamics in Lake Ontario. Deepwater Sculpin Myoxocephalus thompsonii, considered rare or absent from Lake Ontario for 30 years, have generally increased over the past eight years. For the first time since they were caught in this assessment, Deepwater Sculpin density and biomass density estimates declined from the previous year. The 2013 abundance and density estimates for trawls covering the standard depths from 60m to 150m was 0.0001 fish per square meter and 0.0028 grams per square meter. In 2013, very few small (< 80 mm) Deepwater Sculpin were caught and most sculpin were at sites of 150 meters or greater, which is in contrast to previous years when juvenile fish were caught around 80-100 meters. The reduced effort and late seasonal timing of the 2013 assessment make it difficult to have high confidence in declines observed in 2013, however observed Alewife Alosa psuedoharengus abundance increases and

Fluorescent in situ hybridization (FISH) on corneal impression cytology specimens (CICS): study of epithelial cell survival after keratoplasty.

PubMed

Catanese, Muriel; Popovici, Cornel; Proust, Hélène; Hoffart, Louis; Matonti, Frédéric; Cochereau, Isabelle; Conrath, John; Gabison, Eric E

2011-02-22

To assess corneal epithelial cell survival after keratoplasty. Corneal impression cytology (CIC) was performed on sex-mismatched corneal transplants. Fluorescent in situ hybridization (FISH) with sex chromosome-specific probes was performed to identify epithelial cell mosaicism and therefore allocate the donor or recipient origin of the cells. Twenty-four samples of corneal epithelial cells derived from 21 transplanted patients were analyzed. All patients received post-operative treatment using dexamethasone eye drops, with progressive tapering over 18 months, and nine patients also received 2% cyclosporine eye drops. Out of the 24 samples reaching quality criteria, sex mosaicism was found in 13, demonstrating the presence of donor-derived cells at the center of the graft for at least 211 days post keratoplasty. Kaplan-Meier analysis established a median survival of donor corneal epithelial cells of 385 days. Although not statistically significant, the disappearance of donor cells seemed to be delayed and the average number of persistent cells appeared to be greater when 2% cyclosporine was used topically as an additional immunosuppressive therapy. The combination of corneal impressions and FISH analysis is a valuable tool with negligible side effects to investigate the presence of epithelial cell mosaicism in sex-mismatched donor transplants. Epithelial cells survived at the center of the graft with a median survival of more than one year, suggesting slower epithelial turnover than previously described.

Clinical significance of detecting circulating tumor cells in colorectal cancer using subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH)

PubMed Central

Shen, Zhen; Jing, Yan; Lu, Haibo; Li, Heng; Yang, Xiaoye; Cui, Xiangbin; Li, Yuqing; Lou, Zheng; Liu, Peng; Zhang, Cun; Zhang, Wei

2017-01-01

Circulating tumor cells (CTC) are useful in early detection of colorectal cancer. This study described a newly developed platform, integrated subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH), to assess CTCs in colorectal cancer. CTCs were detected by SE-iFISH in 40 of 44 preoperative colorectal cancer patients, and yielded a sensitivity of 90.9%, which was significantly higher than CellSearch system (90.9% vs. 43.2%, P=0.033). No significant association was found between tumor stage, survival and preoperative CTC number. CTCs were detected in 10 colorectal cancer patients one week after surgery; seven patients with decreased CTC numbers (compared with preoperative CTC number) were free of recurrence; whereas two of the three patients with increased CTC numbers had tumor recurrence. Moreover, CTCs were detected in 34 colorectal cancer patients three months after surgery; patients with CTC<2 at three months after surgery had significantly longer Progression Free Survival than those with CTC>=2 (P=0.019); patients with decreased CTC number (compared with preoperative CTC number) had significantly longer Progression Free Survival than those with increased CTC number (P=0.003). In conclusion, CTCs could be detected in various stages of colorectal cancer using SE-iFISH. Dynamic monitoring of CTC numbers could predict recurrence and prognosis. PMID:28423493

Identification of the centromeric repeat in the threespine stickleback fish (Gasterosteus aculeatus).

PubMed

Cech, Jennifer N; Peichel, Catherine L

2015-12-01

Centromere sequences exist as gaps in many genome assemblies due to their repetitive nature. Here we take an unbiased approach utilizing centromere protein A (CENP-A) chomatin immunoprecipitation followed by high-throughput sequencing to identify the centromeric repeat sequence in the threespine stickleback fish (Gasterosteus aculeatus). A 186-bp, AT-rich repeat was validated as centromeric using both fluorescence in situ hybridization (FISH) and immunofluorescence combined with FISH (IF-FISH) on interphase nuclei and metaphase spreads. This repeat hybridizes strongly to the centromere on all chromosomes, with the exception of weak hybridization to the Y chromosome. Together, our work provides the first validated sequence information for the threespine stickleback centromere.

Assessing potential health risks to fish and humans using mercury concentrations in inland fish from across western Canada and the United States

USGS Publications Warehouse

Lepak, Jesse M.; Hooten, Mevin B.; Eagles-Smith, Collin A.; Tate, Michael T.; Lutz, Michelle A.; Ackerman, Joshua T.; Willacker, James J.; Jackson, Allyson K.; Evers, David C.; Wiener, James G.; Pritz, Colleen Flanagan; Davis, Jay

2016-01-01

Fish represent high quality protein and nutrient sources, but Hg contamination is ubiquitous in aquatic ecosystems and can pose health risks to fish and their consumers. Potential health risks posed to fish and humans by Hg contamination in fish were assessed in western Canada and the United States. A large compilation of inland fish Hg concentrations was evaluated in terms of potential health risk to the fish themselves, health risk to predatory fish that consume Hg contaminated fish, and to humans that consume Hg contaminated fish. The probability that a fish collected from a given location would exceed a Hg concentration benchmark relevant to a health risk was calculated. These exceedance probabilities and their associated uncertainties were characterized for fish of multiple size classes at multiple health-relevant benchmarks. The approach was novel and allowed for the assessment of the potential for deleterious health effects in fish and humans associated with Hg contamination in fish across this broad study area. Exceedance probabilities were relatively common at low Hg concentration benchmarks, particularly for fish in larger size classes. Specifically, median exceedances for the largest size classes of fish evaluated at the lowest Hg concentration benchmarks were 0.73 (potential health risks to fish themselves), 0.90 (potential health risk to predatory fish that consume Hg contaminated fish), and 0.97 (potential for restricted fish consumption by humans), but diminished to essentially zero at the highest benchmarks and smallest fish size classes. Exceedances of benchmarks are likely to have deleterious health effects on fish and limit recommended amounts of fish humans consume in western Canada and the United States. Results presented here are not intended to subvert or replace local fish Hg data or consumption advice, but provide a basis for identifying areas of potential health risk and developing more focused future research and monitoring efforts.

Can environmental impact assessments alone conserve freshwater fish biota? Review of the Chilean experience

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lacy, Shaw Nozaki, E-mail: shaw.lacy@gmail.com; Departmento de Ecosistemas y Medio Ambiente, Pontificia Universidad Católica de Chile, Vicuña Mackenna 4860, Macul; Centro Interdisciplinario de Cambio Global, Vicuña Mackenna 4860, Macul

Chile was one of many countries that initiated environmental impact assessments in the 1990s, and has relied on their use for species conservation and territorial planning without the use of larger-scale environmental and ecological planning. The capacity of Chile's environmental impact assessment system (SEIA) to evaluate resident freshwater fishes and the potential impacts of water projects and aquaculture activities – two categories of projects that create direct threats to freshwater fishes – are assessed. Of the 3997 such submissions to the SEIA, only 0.6% conducted any freshwater fish assessment, and only 0.1% conducted any quantitative assessment of expected impacts frommore » the associated project. The small number of assessments was characterized by poor study design, inconsistent sampling methodology, and species misidentification. Traditional assessments failed to include freshwater fish ecology in the general assessment framework. The new strategic environmental evaluation system only underscores the need for vastly improved field sampling protocols and assessment methodologies.« less

LARGE RIVER ASSESSMENT METHODS FOR BENTHIC MACROINVERTEBRATES AND FISH

EPA Science Inventory

Multiple projects are currently underway to increase our understanding of the varying results of different sampling methods and designs used for the biological assessment and monitoring of large (boatable) rivers. Studies include methods used to assess fish, benthic macroinverte...

Using posts to an online social network to assess fishing effort

USGS Publications Warehouse

Martin, Dustin R.; Chizinski, Christopher J.; Eskridge, Kent M.; Pope, Kevin L

2014-01-01

Fisheries management has evolved from reservoir to watershed management, creating a need to simultaneously gather information within and across interacting reservoirs. However, costs to gather information on the fishing effort on multiple reservoirs using traditional creel methodology are often prohibitive. Angler posts about reservoirs online provide a unique medium to test hypotheses on the distribution of fishing pressure. We show that the activity on an online fishing social network is related to fishing effort and can be used to facilitate management goals. We searched the Nebraska Fish and Game Association Fishing Forum for all references from April 2009 to December 2010 to 19 reservoirs that comprise the Salt Valley regional fishery in southeastern Nebraska. The number of posts was positively related to monthly fishing effort on a regional scale, with individual reservoirs having the most annual posts also having the most annual fishing effort. Furthermore, this relationship held temporally. Online fishing social networks provide the potential to assess effort on larger spatial scales than currently feasible.

Fish pass assessment by remote control: a novel framework for quantifying the hydraulics at fish pass entrances

NASA Astrophysics Data System (ADS)

Kriechbaumer, Thomas; Blackburn, Kim; Gill, Andrew; Breckon, Toby; Everard, Nick; Wright, Ros; Rivas Casado, Monica

2014-05-01

Fragmentation of aquatic habitats can lead to the extinction of migratory fish species with severe negative consequences at the ecosystem level and thus opposes the target of good ecological status of rivers defined in the EU Water Framework Directive (WFD). In the UK, the implementation of the EU WFD requires investments in fish pass facilities of estimated 532 million GBP (i.e. 639 million Euros) until 2027 to ensure fish passage at around 3,000 barriers considered critical. Hundreds of passes have been installed in the past. However, monitoring studies of fish passes around the world indicate that on average less than half of the fish attempting to pass such facilities are actually successful. There is a need for frameworks that allow the rapid identification of facilities that are biologically effective and those that require enhancement. Although there are many environmental characteristics that can affect fish passage success, past research suggests that variations in hydrodynamic conditions, reflected in water velocities, velocity gradients and turbulences, are the major cues that fish use to seek migration pathways in rivers. This paper presents the first steps taken in the development of a framework for the rapid field-based quantification of the hydraulic conditions downstream of fish passes and the assessment of the attractivity of fish passes for salmonids and coarse fish in UK rivers. For this purpose, a small-sized remote control platform carrying an acoustic Doppler current profiler (ADCP), a GPS unit, a stereo camera and an inertial measurement unit has been developed. The large amount of data on water velocities and depths measured by the ADCP within relatively short time is used to quantify the spatial and temporal distribution of water velocities. By matching these hydraulic features with known preferences of migratory fish, it is attempted to identify likely migration routes and aggregation areas at barriers as well as hydraulic features that

Evaluation of HER-2/neu status in breast cancer specimens using immunohistochemistry (IHC) & fluorescence in-situ hybridization (FISH) assay.

PubMed

Goud, Kalal Iravathy; Dayakar, Seetha; Vijayalaxmi, Kolanupaka; Babu, Saidam Jangu; Reddy, P Vijay Anand

2012-03-01

Fluorescence in situ hybridization (FISH) is increasingly being recognized as the most accurate and predictive test for HER 2/neu gene amplification and response to therapy in breast cancer. In the present study we investigated HER-2/neu gene amplification by FISH in breast carcinoma tissue specimens and compared the results with that of immunohistochemical (IHC) analysis. A total of 90 breast carcinoma tissue samples were used for immunohistochemical (IHC) and FISH analysis. IHC was performed by using mouse monoclonal antibody to the intracellular domain of HER-2/neu protein. Each slide was scored in a blinded fashion by two pathologists according to the manufacturer's recommended criteria. FISH analysis was performed on paraffin embedded breast tumour tissue sections. The polysomy for centromere 17 (Spec green signal) was read as green signals less than 4 as moderate polysomy, and more than 4 as highly polysomy. Thirty of the 90 patients had negative results by IHC and FISH. Of the 28 patients with the score of 2+ by IHC, 20 were FISH positive for HER-2/neu gene amplification, three were FISH negative and five patients showed equivocal (1.8-2.2) results by FISH. These five cases were retested for IHC and FISH on different paraffin embedded tissue blocks, and all five were found positive for HER-2/neu gene amplification. Twenty five patients with the score of 3+ by IHC were FISH positive for HER-2/neu gene amplification (>2.2). Seven cases with the score of 3+ by IHC were FISH negative for HER-2/neu gene amplification (>2.2), and showed polysomy of chromosome number 17 high polysomy > 4. Our results indicated that HER-2/neu status by FISH should be performed in all cases of breast tumour with a 2+ score by IHC. Cases demonstrating a 3+ score by IHC may be subjected to FISH to rule out polysomy of chromosome 17 which could be falsely interpreted as HER-2/neu overexpression by IHC analysis. There is also a need for establishing a clinically validated cut-off value

Evaluation of HER-2/neu status in breast cancer specimens using immunohistochemistry (IHC) & fluorescence in-situ hybridization (FISH) assay

PubMed Central

Goud, Kalal Iravathy; Dayakar, Seetha; Vijayalaxmi, Kolanupaka; Babu, Saidam Jangu; Vijay, Anand Reddy P.

2012-01-01

Background & objectives: Fluorescence in situ hybridization (FISH) is increasingly being recognized as the most accurate and predictive test for HER2/neu gene amplification and response to therapy in breast cancer. In the present study we investigated HER-2/neu gene amplification by FISH in breast carcinoma tissue specimens and compared the results with that of immunohistochemical (IHC) analysis. Methods: A total of 90 breast carcinoma tissue samples were used for immunohistochemical (IHC) and FISH analysis. IHC was performed by using mouse monoclonal antibody to the intracellular domain of HER-2/neu protein. Each slide was scored in a blinded fashion by two pathologists according to the manufacturer's recommended criteria. FISH analysis was performed on paraffin embedded breast tumour tissue sections. The polysomy for centromere 17 (Spec green signal) was read as green signals less than 4 as moderate polysomy, and more than 4 as highly polysomy. Results: Thirty of the 90 patients had negative results by IHC and FISH. Of the 28 patients with the score of 2+ by IHC, 20 were FISH positive for HER-2/neu gene amplification, three were FISH negative and five patients showed equivocal (1.8-2.2) results by FISH. These five cases were retested for IHC and FISH on different paraffin embedded tissue blocks, and all five were found positive for HER-2/neu gene amplification. Twenty five patients with the score of 3+ by IHC were FISH positive for HER-2/neu gene amplification (>2.2). Seven cases with the score of 3+ by IHC were FISH negative for HER-2/neu gene amplification (>2.2), and showed polysomy of chromosome number 17 high polysomy > 4. Interpretation & conclusions: Our results indicated that HER-2/neu status by FISH should be performed in all cases of breast tumour with a 2+ score by IHC. Cases demonstrating a 3+ score by IHC may be subjected to FISH to rule out polysomy of chromosome 17 which could be falsely interpreted as HER-2/neu overexpression by IHC analysis

Population cage experiments with a vertebrate: The temporal demography and cytonuclear genetics of hybridization on Gambusia fishes

USGS Publications Warehouse

Scribner, Kim T.; Avise, John C.

1994-01-01

The dynamics of mitochondrial and multilocus nuclear genotypic frequencies were monitored for 2 yr in experimental populations established with equal numbers of two poeciliid fishes (Gambusia affinis and Gambusia holbrooki) that hybridize naturally in the southeastern United States. In replicated "small-pool" populations (experiment I), 1018 sampled individuals at six time periods revealed an initial flush of hybridization, followed by a rapid decline in frequencies of G. affinis nuclear and mitochondrial alleles over 64 wk. Decay of gametic and cytonuclear disequilibria differed from expectations under random mating as well as under a model of assortative mating involving empirically estimated mating propensities. In two replicate "large-pond" populations (experiment II), 841 sampled individuals across four reproductive cohorts revealed lower initial frequencies of F1 hybrids than in experiment I, but again G. holbrooki alleles achieved high frequencies over four generations (72 wk). Thus, evolution within experimental Gambusia hybrid populations can be extremely rapid, resulting in consistent loss of G. affinis nuclear and cytoplasmic alleles. Concordance in results between experiments and across genetic markers suggests strong directional selection favoring G. holbrooki genotypes. Results are interpreted in light of previous reports of genotype-specific differences in life-history traits, reproductive ecology, patterns of recruitment, and size-specific mortality, and in the context of patterns of introgression previously studied indirectly from spatial observations on cytonuclear genotypes in natural Gambusia populations.

Potential for bias in using hybrids between common carp (Cyprinus carpio) and goldfish (Carassius auratus) in endocrine studies: a first report of hybrids in Lake Mead, Nevada, U.S.A

USGS Publications Warehouse

Goodbred, Steven L.; Patino, Reynaldo; Orsak, Erik; Sharma, Prakash; Ruessler, Shane

2013-01-01

During a 2008 study to assess endocrine and reproductive health of common carp (Cyprinus carpio) in Lake Mead, Nevada (U.S.A.) we identified two fish, one male and one female, as hybrids with goldfish (Carassius auratus) based on morphology, lateral line scale count, and lack of anterior barbels. Gross examination of the female hybrid ovaries indicated presence of vitellogenic ovarian follicles; whereas histological evaluation of the male hybrid testes showed lobule-like structures with open lumens but without germ cells, suggesting it was sterile. Because common carp/goldfish hybrids are more susceptible to gonadal tumors and may have different endocrine profiles than common carp, researchers using common carp as a model for endocrine/reproductive studies should be aware of the possible presence of hybrids.

A high-resolution cat radiation hybrid and integrated FISH mapping resource for phylogenomic studies across Felidae.

PubMed

Davis, Brian W; Raudsepp, Terje; Pearks Wilkerson, Alison J; Agarwala, Richa; Schäffer, Alejandro A; Houck, Marlys; Chowdhary, Bhanu P; Murphy, William J

2009-04-01

We describe the construction of a high-resolution radiation hybrid (RH) map of the domestic cat genome, which includes 2662 markers, translating to an estimated average intermarker distance of 939 kilobases (kb). Targeted marker selection utilized the recent feline 1.9x genome assembly, concentrating on regions of low marker density on feline autosomes and the X chromosome, in addition to regions flanking interspecies chromosomal breakpoints. Average gap (breakpoint) size between cat-human ordered conserved segments is less than 900 kb. The map was used for a fine-scale comparison of conserved syntenic blocks with the human and canine genomes. Corroborative fluorescence in situ hybridization (FISH) data were generated using 129 domestic cat BAC clones as probes, providing independent confirmation of the long-range correctness of the map. Cross-species hybridization of BAC probes on divergent felids from the genera Profelis (serval) and Panthera (snow leopard) provides further evidence for karyotypic conservation within felids, and demonstrates the utility of such probes for future studies of chromosome evolution within the cat family and in related carnivores. The integrated map constitutes a comprehensive framework for identifying genes controlling feline phenotypes of interest, and to aid in assembly of a higher coverage feline genome sequence.

A High-Resolution Cat Radiation Hybrid and Integrated FISH Mapping Resource for Phylogenomic Studies across Felidae

PubMed Central

Davis, Brian W.; Raudsepp, Terje; Wilkerson, Alison J. Pearks; Agarwala, Richa; Schäffer, Alejandro A.; Houck, Marlys; Ryder, Oliver A.; Chowdhdary, Bhanu P.; Murphy, William J.

2008-01-01

We describe the construction of a high-resolution radiation hybrid (RH) map of the domestic cat genome, which includes 2,662 markers, translating to an estimated average intermarker distance of 939 kilobases (Kb). Targeted marker selection utilized the recent feline 1.9x genome assembly, concentrating on regions of low marker density on feline autosomes and the X chromosome, in addition to regions flanking interspecies chromosomal breakpoints. Average gap (breakpoint) size between cat-human ordered conserved segments is less than 900 Kb. The map was used for a fine-scale comparison of conserved syntenic blocks with the human and canine genomes. Corroborative fluorescence in situ hybridization (FISH) data were generated using 129 domestic cat BAC-clones as probes, providing independent confirmation of the long-range correctness of the map. Cross-species hybridization of BAC probes on divergent felids from the genera Profelis (serval) and Panthera (snow leopard) provides further evidence for karyotypic conservation within felids, and demonstrates the utility of such probes for future studies of chromosome evolution within the cat family and in related carnivores. The integrated map constitutes a comprehensive framework for identifying genes controlling feline phenotypes of interest, and to aid in assembly of a higher coverage feline genome sequence. PMID:18951970

Identification of diazotrophic microorganisms in marine sediment via fluorescence in situ hybridization coupled to nanoscale secondary ion mass spectrometry (FISH-NanoSIMS).

PubMed

Dekas, Anne E; Orphan, Victoria J

2011-01-01

Growing appreciation for the biogeochemical significance of uncultured microorganisms is changing the focus of environmental microbiology. Techniques designed to investigate microbial metabolism in situ are increasingly popular, from mRNA-targeted fluorescence in situ hybridization (FISH) to the "-omics" revolution, including metagenomics, transcriptomics, and proteomics. Recently, the coupling of FISH with nanometer-scale secondary ion mass spectrometry (NanoSIMS) has taken this movement in a new direction, allowing single-cell metabolic analysis of uncultured microbial phylogenic groups. The main advantage of FISH-NanoSIMS over previous noncultivation-based techniques to probe metabolism is its ability to directly link 16S rRNA phylogenetic identity to metabolic function. In the following chapter, we describe the procedures necessary to identify nitrogen-fixing microbes within marine sediment via FISH-NanoSIMS, using our work on nitrogen fixation by uncultured deep-sea methane-consuming archaea as a case study. Copyright © 2011 Elsevier Inc. All rights reserved.

Fish Alternatives in Environmental Risk Assessment: Overview of the Current Landscape

EPA Science Inventory

The need for alternative testing strategies has recently expanded into the realm of environmental risk assessment leading to the development of new alternatives to standard aquatic vertebrate testing such as the OECD 203 acute fish toxicity test. The fish embryo test (FET) is one...

Techno-economic assessment of a hybrid solar receiver and combustor

NASA Astrophysics Data System (ADS)

Lim, Jin Han; Nathan, Graham; Dally, Bassam; Chinnici, Alfonso

2016-05-01

A techno-economic analysis is performed to compare two different configurations of hybrid solar thermal systems with fossil fuel backup to provide continuous electricity output. The assessment compares a Hybrid Solar Receiver Combustor (HSRC), in which the functions of a solar cavity receiver and a combustor are integrated into a single device with a reference conventional solar thermal system using a regular solar cavity receiver with a backup boiler, termed the Solar Gas Hybrid (SGH). The benefits of the integration is assessed by varying the size of the storage capacity and heliostat field while maintaining the same overall thermal input to the power block.

Environmental risk assessment of alkylphenols from offshore produced water on fish reproduction.

PubMed

Beyer, Jonny; Myhre, Lars Petter; Sundt, Rolf C; Meier, Sonnich; Tollefsen, Knut-Erik; Vabø, Rune; Klungsøyr, Jarle; Sanni, Steinar

2012-04-01

Concern has been raised over whether environmental release of alkylphenols (AP) in produced water (PW) discharges from the offshore oil industry could impose a risk to the reproduction of fish stocks in the North Sea. An environmental risk assessment (ERA) was performed to determine if environmental exposure to PW APs in North Sea fish populations is likely to be high enough to give effects on reproduction endpoints. The DREAM (Dose related Risk and Effect Assessment Model) software was used in the study and the inputs to the ERA model included PW discharge data, fate information of PW plumes, fish distribution information, as well as uptake and elimination information of PW APs. Toxicodynamic data from effect studies with Atlantic cod (Gadus morhua) exposed to APs were used to establish a conservative environmental risk threshold value for AP concentration in seawater. By using the DREAM software to 1) identify the areas of highest potential risk and 2) integrate fish movement and uptake/elimination rates of APs for the chosen areas we found that the environmental exposure of fish to APs from PW is most likely too low to affect reproduction in wild populations of fish in the North Sea. The implications related to risk management of offshore PW and uncertainties in the risk assessment performed are discussed. Copyright © 2011 Elsevier Ltd. All rights reserved.

Cortisol Release in Response to UVB Exposure in Xiphophorus Fish

PubMed Central

Contreras, Adam J.; Boswell, Mikki; Downs, Kevin P.; Pasquali, Amanda; Walter, Ronald B.

2014-01-01

Xiphophorus fishes are comprised of 26 known species. Interspecies hybridization between select species has been utilized to produce experimental models to study melanoma development. Xiphophorus melanoma induction protocols utilize ultraviolet light (UVB) to induce DNA damage and associated downstream tumorigenesis. However, the impact of induced stress caused by the UVB treatment of the experimental animals undergoing tumor induction protocols has not been assessed. Stress is an adaptive physiological response to excessive or unpredictable environmental stimuli. The stress response in fishes may be measured by assay of cortisol released into the water. Here, we present results from investigations of stress response during experimental treatment and UVB exposure in X. maculatus Jp 163 B, X. couchianus, and F1 interspecies hybrids produced from the mating X. maculatus Jp 163 B x X. couchianus. Overall, cortisol release rates for males and females after UVB exposure showed no statistical differences. At lower UVB doses (8 and 16 kJ/m2), X. couchianus exhibited 2 fold higher levels of DNA damage then either X. maculatus or the F1 hybrid. However, based on cortisol release rates, none of the fish types tested induced a primary stress response at the UVB lower doses (8 and 16 kJ/m2). In contrast, at a very high UVB dose (32 kJ/m2) both X. maculatus and the F1 hybrid showed a 5 fold increase in cortisol release rate. To determine the effect of pigmentation on UVB induced stress, wild type and albino X. hellerii were exposed to UVB (32 kJ/m2). Albino X. hellerii exhibited 3.7 fold increase in cortisol release while wild type X. hellerii did not exhibit a significant cortisol response to UVB. Overall, the data suggest the rather low UVB doses often employed in tumour induction protocols do not induce a primary stress response in Xiphophorus fishes. PMID:24625568

Assessment of the potential of hybrid vehicles: Summary

NASA Technical Reports Server (NTRS)

Surber, F. T.

1980-01-01

The potential of hybrid vehicles as a replacement of the conventional gasoline or diesel fueled internal combustion engine vehicle within the next 20 to 30 years, was assessed. Hybrid vehicle designs and applications which are technically and economically viable were studied to determine if reductions in petroleum usage were large enough to warrant major expenditures of research and development funds. Critical technical areas where research and development can be most usefully concentrated were identified.

Computerized analysis of cytology and fluorescence in situ hybridization (FISH) in induced sputum for lung cancer detection.

PubMed

Guber, Alexander; Greif, Joel; Rona, Roni; Fireman, Elizabeth; Madi, Lea; Kaplan, Tal; Yemini, Zipi; Gottfried, Maya; Katz, Ruth L; Daniely, Michal

2010-10-25

Lung cancer results from a multistep process, whereby genetic and epigenetic alterations lead to a malignant phenotype. Somatic mutations, deletions, and amplifications can be detected in the tumor itself, but they can also be found in histologically normal bronchial epithelium as a result of field cancerization. The present feasibility study describes a computer-assisted analysis of induced sputum employing morphology and fluorescence in situ hybridization (target-FISH), using 2 biomarkers located at chromosomes 3p22.1 and 10q22.3. Induced sputum samples were collected using a standardized protocol from 12 patients with lung cancer and from 15 healthy, nonsmoking controls. We used an automated scanning system that allows consecutive scans of morphology and FISH of the same slide. Cells derived for the lower airways were analyzed for the presence of genetic alterations in the 3p22.1 and 10q22.3 loci. The cutoff for a positive diagnosis was defined as >4% of cells showing genetic alterations. Eleven of 12 lung cancer patients and 12 of 15 controls were identified correctly, giving an overall sensitivity and specificity of 91.66% and 80%, respectively. This study describes a new technology for detecting lung cancer noninvasively in induced sputum via a combination of morphology and FISH analysis (target-FISH) using computer-assisted technology. This approach may potentially be utilized for mass screening of high-risk populations. © 2010 American Cancer Society.

Production of F1 interspecies hybrid offspring with cryopreserved sperm from a live-bearing fish, the swordtail Xiphophorus helleri.

PubMed

Yang, Huiping; Hazlewood, Leona; Heater, Sheila J; Guerrero, Paula A; Walter, Ronald B; Tiersch, Terrence R

2007-03-01

Despite study of sperm cryopreservation in more than 200 fish species, production of broods from cryopreserved sperm in live-bearing fish has not been demonstrated. This has not been due to a lack of effort, but instead is a result of the unique morphology, biology, and biochemistry of reproduction in viviparous fishes. For example, sperm of Xiphophorus helleri have a cylindrical nucleus, can swim for days after being activated, have glycolytic capabilities, and can reside in the female reproduction tract for months before fertilization. These traits are not found in fishes with external fertilization. The long-standing research use of the genus Xiphophorus has led to development of over 60 pedigreed lines among the 26 species maintained around the world. These species and lines serve as contemporary models in medical research, although they must be maintained as live populations. Previous attempts at establishing sperm cryopreservation protocols for Xiphophorus have not produced live young. To address this we have been studying the parameters surrounding cryobiology of Xiphophorus sperm and applying this information to an improved understanding of internal fertilization and reproduction. Here we report the first successful fertilization and offspring production by cryopreserved sperm in any live-bearing fish. This claim is supported by our use of artificial insemination between two species that yield distinct hybrid offspring to verify paternity via cryopreserved sperm. We provide a practical approach for preservation of valuable genetic resources from live-bearing fish species, a group that is rapidly being lost due to destruction of native habitats.

Food wastes as fish feeds for polyculture of low-trophic-level fish: bioaccumulation and health risk assessments of heavy metals in the cultured fish.

PubMed

Cheng, Zhang; Lam, Cheung-Lung; Mo, Wing-Yin; Nie, Xiang-Ping; Choi, Wai-Ming; Man, Yu-Bon; Wong, Ming-Hung

2016-04-01

The major purpose of this study was to use different types of food wastes which serve as the major sources of protein to replace the fish meal used in fish feeds to produce quality fish. Two types of food waste-based feed pellets FW A (with cereals) and FW B (with cereals and meat products) and the commercial feed Jinfeng® were used to culture fingerlings of three low-trophic-level fish species: bighead carp, grass carp, and mud carp (in the ratio of 1:3:1) for 1 year period in the Sha Tau Kok Organic Farm in Hong Kong. Heavy metal concentrations in all of the fish species fed with food waste pellets and commercial pellets in Sha Tau Kok fish ponds were all below the local and international maximum permissible levels in food. Health risk assessments indicated that human consumption of the fish fed with food waste feed pellets was safe for the Hong Kong residents. The present results revealed that recycling of food waste for cultivating low-trophic-level fish (mainly herbivores and detritus feeders) is feasible, and at the same time will ease the disposal pressure of food waste, a common problem of densely populated cities like Hong Kong.

5-bp Classical Satellite DNA Loci from Chromosome-1 Instability in Cervical Neoplasia Detected by DNA Breakage Detection/Fluorescence in Situ Hybridization (DBD-FISH).

PubMed

Cortés-Gutiérrez, Elva I; Ortíz-Hernández, Brenda L; Dávila-Rodríguez, Martha I; Cerda-Flores, Ricardo M; Fernández, José Luis; López-Fernández, Carmen; Gosálvez, Jaime

2013-02-19

We aimed to evaluate the association between the progressive stages of cervical neoplasia and DNA damage in 5-bp classical satellite DNA sequences from chromosome-1 in cervical epithelium and in peripheral blood lymphocytes using DNA breakage detection/fluorescence in situ hybridization (DBD-FISH). A hospital-based unmatched case-control study was conducted in 2011 with a sample of 30 women grouped according to disease stage and selected according to histological diagnosis; 10 with low-grade squamous intraepithelial lesions (LG-SIL), 10 with high-grade SIL (HG-SIL), and 10 with no cervical lesions, from the Unidad Medica de Alta Especialidad of The Mexican Social Security Institute, IMSS, Mexico. Specific chromosome damage levels in 5-bp classical satellite DNA sequences from chromosome-1 were evaluated in cervical epithelium and peripheral blood lymphocytes using the DBD-FISH technique. Whole-genome DNA hybridization was used as a reference for the level of damage. Results of Kruskal-Wallis test showed a significant increase according to neoplastic development in both tissues. The instability of 5-bp classical satellite DNA sequences from chromosome-1 was evidenced using chromosome-orientation FISH. In conclusion, we suggest that the progression to malignant transformation involves an increase in the instability of 5-bp classical satellite DNA sequences from chromosome-1.

Predictive models for fish assemblages in eastern USA streams: implications for assessing biodiversity

USGS Publications Warehouse

Meador, Michael R.; Carlisle, Daren M.

2009-01-01

Management and conservation of aquatic systems require the ability to assess biological conditions and identify changes in biodiversity. Predictive models for fish assemblages were constructed to assess biological condition and changes in biodiversity for streams sampled in the eastern United States as part of the U.S. Geological Survey's National Water Quality Assessment Program. Separate predictive models were developed for northern and southern regions. Reference sites were designated using land cover and local professional judgment. Taxonomic completeness was quantified based on the ratio of the number of observed native fish species expected to occur to the number of expected native fish species. Models for both regions accurately predicted fish species composition at reference sites with relatively high precision and low bias. In general, species that occurred less frequently than expected (decreasers) tended to prefer riffle areas and larger substrates, such as gravel and cobble, whereas increaser species (occurring more frequently than expected) tended to prefer pools, backwater areas, and vegetated and sand substrates. In the north, the percentage of species identified as increasers and the percentage identified as decreasers were equal, whereas in the south nearly two-thirds of the species examined were identified as decreasers. Predictive models of fish species can provide a standardized indicator for consistent assessments of biological condition at varying spatial scales and critical information for an improved understanding of fish species that are potentially at risk of loss with changing water quality conditions.

Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

PubMed

Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

2015-06-01

In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis.

Clarithromycin resistance of Helicobacter pylori strains isolated from children' gastric antrum and fundus as assessed by fluorescent in-situ hybridization and culture on four-sector agar plates.

PubMed

Caristo, Elisa; Parola, Andrea; Rapa, Anna; Vivenza, Daniela; Raselli, Barbara; Dondi, Elena; Boldorini, Renzo; Oderda, Giuseppina

2008-12-01

To assess validity of culture on four-sector agar plates and fluorescent in-situ hybridization (FISH) test, and clarithromycin resistance rate in Helicobacter pylori strains isolated from children in the last 10 years. In the last 5 years, gastric biopsy specimens from antrum and fundus were taken from 89 consecutive children (median age 9 years) with H. pylori gastritis and from 21 controls. Culture was performed on 176 gastric biopsies (89 from antrum, 87 from fundus) on four-sector agar plates, and FISH test with DNA ProbeMix. After its validity was evaluated, FISH test was applied on additional 119 biopsies from 68 children (68 from the antrum, 51 from the fundus) stored in the Pathology archive in the previous 5 years. Culture was positive in 157 of 176 biopsies (sensitivity: 89.2%, 95% confidence interval (CI) 85-94). In 33 of 89 children (37%) resistant strains were found in one or both gastric sites. FISH test was positive in 148 of 176 biopsies from infected children (sensitivity 84.1%, 95%CI 79-89) and in none of 42 biopsies from controls (specificity 100%). When applied on archive biopsies, FISH test was positive in 96 of 119 (80.7%, 95%CI 74-88). Total children harboring resistant strains in the last 10 years, as assessed by FISH test, were 66 of 157 (42%). Mixed infection with both sensitive and resistant strains were found in 40 children (25%) and in 12 of them resistant strains were in the fundus only. Culture on four-sector agar plates and FISH test had a high sensitivity and specificity and showed co-presence of sensitive and resistant strains. In one-third of children with mixed infection, the resistant strains were in the fundus only. Clarithromycin resistance should be assessed in biopsies both from the antrum and the fundus, utilizing antral biopsies only can underestimate its prevalence.

Reef Fish Survey Techniques: Assessing the Potential for Standardizing Methodologies.

PubMed

Caldwell, Zachary R; Zgliczynski, Brian J; Williams, Gareth J; Sandin, Stuart A

2016-01-01

Dramatic changes in populations of fishes living on coral reefs have been documented globally and, in response, the research community has initiated efforts to assess and monitor reef fish assemblages. A variety of visual census techniques are employed, however results are often incomparable due to differential methodological performance. Although comparability of data may promote improved assessment of fish populations, and thus management of often critically important nearshore fisheries, to date no standardized and agreed-upon survey method has emerged. This study describes the use of methods across the research community and identifies potential drivers of method selection. An online survey was distributed to researchers from academic, governmental, and non-governmental organizations internationally. Although many methods were identified, 89% of survey-based projects employed one of three methods-belt transect, stationary point count, and some variation of the timed swim method. The selection of survey method was independent of the research design (i.e., assessment goal) and region of study, but was related to the researcher's home institution. While some researchers expressed willingness to modify their current survey protocols to more standardized protocols (76%), their willingness decreased when methodologies were tied to long-term datasets spanning five or more years. Willingness to modify current methodologies was also less common among academic researchers than resource managers. By understanding both the current application of methods and the reported motivations for method selection, we hope to focus discussions towards increasing the comparability of quantitative reef fish survey data.

Dealing With Uncertainty When Assessing Fish Passage Through Culvert Road Crossings

NASA Astrophysics Data System (ADS)

Anderson, Gregory B.; Freeman, Mary C.; Freeman, Byron J.; Straight, Carrie A.; Hagler, Megan M.; Peterson, James T.

2012-09-01

Assessing the passage of aquatic organisms through culvert road crossings has become increasingly common in efforts to restore stream habitat. Several federal and state agencies and local stakeholders have adopted assessment approaches based on literature-derived criteria for culvert impassability. However, criteria differ and are typically specific to larger-bodied fishes. In an analysis to prioritize culverts for remediation to benefit imperiled, small-bodied fishes in the Upper Coosa River system in the southeastern United States, we assessed the sensitivity of prioritization to the use of differing but plausible criteria for culvert impassability. Using measurements at 256 road crossings, we assessed culvert impassability using four alternative criteria sets represented in Bayesian belief networks. Two criteria sets scored culverts as either passable or impassable based on alternative thresholds of culvert characteristics (outlet elevation, baseflow water velocity). Two additional criteria sets incorporated uncertainty concerning ability of small-bodied fishes to pass through culverts and estimated a probability of culvert impassability. To prioritize culverts for remediation, we combined estimated culvert impassability with culvert position in the stream network relative to other barriers to compute prospective gain in connected stream habitat for the target fish species. Although four culverts ranked highly for remediation regardless of which criteria were used to assess impassability, other culverts differed widely in priority depending on criteria. Our results emphasize the value of explicitly incorporating uncertainty into criteria underlying remediation decisions. Comparing outcomes among alternative, plausible criteria may also help to identify research most needed to narrow management uncertainty.

Dealing with uncertainty when assessing fish passage through culvert road crossings.

PubMed

Anderson, Gregory B; Freeman, Mary C; Freeman, Byron J; Straight, Carrie A; Hagler, Megan M; Peterson, James T

2012-09-01

Assessing the passage of aquatic organisms through culvert road crossings has become increasingly common in efforts to restore stream habitat. Several federal and state agencies and local stakeholders have adopted assessment approaches based on literature-derived criteria for culvert impassability. However, criteria differ and are typically specific to larger-bodied fishes. In an analysis to prioritize culverts for remediation to benefit imperiled, small-bodied fishes in the Upper Coosa River system in the southeastern United States, we assessed the sensitivity of prioritization to the use of differing but plausible criteria for culvert impassability. Using measurements at 256 road crossings, we assessed culvert impassability using four alternative criteria sets represented in Bayesian belief networks. Two criteria sets scored culverts as either passable or impassable based on alternative thresholds of culvert characteristics (outlet elevation, baseflow water velocity). Two additional criteria sets incorporated uncertainty concerning ability of small-bodied fishes to pass through culverts and estimated a probability of culvert impassability. To prioritize culverts for remediation, we combined estimated culvert impassability with culvert position in the stream network relative to other barriers to compute prospective gain in connected stream habitat for the target fish species. Although four culverts ranked highly for remediation regardless of which criteria were used to assess impassability, other culverts differed widely in priority depending on criteria. Our results emphasize the value of explicitly incorporating uncertainty into criteria underlying remediation decisions. Comparing outcomes among alternative, plausible criteria may also help to identify research most needed to narrow management uncertainty.

Dealing with uncertainty when assessing fish passage through culvert road crossings

USGS Publications Warehouse

Anderson, Gregory B.; Freeman, Mary C.; Freeman, Byron J.; Straight, Carrie A.; Hagler, Megan M.; Peterson, James T.

2012-01-01

Assessing the passage of aquatic organisms through culvert road crossings has become increasingly common in efforts to restore stream habitat. Several federal and state agencies and local stakeholders have adopted assessment approaches based on literature-derived criteria for culvert impassability. However, criteria differ and are typically specific to larger-bodied fishes. In an analysis to prioritize culverts for remediation to benefit imperiled, small-bodied fishes in the Upper Coosa River system in the southeastern United States, we assessed the sensitivity of prioritization to the use of differing but plausible criteria for culvert impassability. Using measurements at 256 road crossings, we assessed culvert impassability using four alternative criteria sets represented in Bayesian belief networks. Two criteria sets scored culverts as either passable or impassable based on alternative thresholds of culvert characteristics (outlet elevation, baseflow water velocity). Two additional criteria sets incorporated uncertainty concerning ability of small-bodied fishes to pass through culverts and estimated a probability of culvert impassability. To prioritize culverts for remediation, we combined estimated culvert impassability with culvert position in the stream network relative to other barriers to compute prospective gain in connected stream habitat for the target fish species. Although four culverts ranked highly for remediation regardless of which criteria were used to assess impassability, other culverts differed widely in priority depending on criteria. Our results emphasize the value of explicitly incorporating uncertainty into criteria underlying remediation decisions. Comparing outcomes among alternative, plausible criteria may also help to identify research most needed to narrow management uncertainty.

Optimization of peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) for the detection of bacteria: The effect of pH, dextran sulfate and probe concentration.

PubMed

Rocha, Rui; Santos, Rita S; Madureira, Pedro; Almeida, Carina; Azevedo, Nuno F

2016-05-20

Fluorescence in situ hybridization (FISH) is a molecular technique widely used for the detection and characterization of microbial populations. FISH is affected by a wide variety of abiotic and biotic variables and the way they interact with each other. This is translated into a wide variability of FISH procedures found in the literature. The aim of this work is to systematically study the effects of pH, dextran sulfate and probe concentration in the FISH protocol, using a general peptide nucleic acid (PNA) probe for the Eubacteria domain. For this, response surface methodology was used to optimize these 3 PNA-FISH parameters for Gram-negative (Escherichia coli and Pseudomonas fluorescens) and Gram-positive species (Listeria innocua, Staphylococcus epidermidis and Bacillus cereus). The obtained results show that a probe concentration higher than 300nM is favorable for both groups. Interestingly, a clear distinction between the two groups regarding the optimal pH and dextran sulfate concentration was found: a high pH (approx. 10), combined with lower dextran sulfate concentration (approx. 2% [w/v]) for Gram-negative species and near-neutral pH (approx. 8), together with higher dextran sulfate concentrations (approx. 10% [w/v]) for Gram-positive species. This behavior seems to result from an interplay between pH and dextran sulfate and their ability to influence probe concentration and diffusion towards the rRNA target. This study shows that, for an optimum hybridization protocol, dextran sulfate and pH should be adjusted according to the target bacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

Bottom trawl assessment of Lake Ontario prey fishes

USGS Publications Warehouse

Weidel, Brian C.; Connerton, Michael J.; Holden, Jeremy

2018-01-01

Managing Lake Ontario fisheries in an ecosystem-context requires prey fish community and population data. Since 1978, multiple annual bottom trawl surveys have quantified prey fish dynamics to inform management relative to published Fish Community Objectives. In 2017, two whole-lake surveys collected 341 bottom trawls (spring: 204, fall: 137), at depths from 8-225m, and captured 751,350 fish from 29 species. Alewife were 90% of the total fish catch while Deepwater Sculpin, Round Goby, and Rainbow Smelt comprised the majority of the remaining total catch (3.8, 3.1, and 1.1% respectively). The adult Alewife abundance index for US waters increased in 2017 relative to 2016, however the index for Canadian waters declined. Adult Alewife condition, assessed by the predicted weight of a 165 mm fish (6.5 inches), declined in 2017 from record high values observed in spring 2016. Spring 2017 Alewife condition was slightly less than the 10-year average, but the fall value was well below the 10-year average, likely due to increased Age-1 Alewife abundance. The Age-1 Alewife abundance index was the highest observed in 40 years, and 8-times higher than the previous year. The Age-1 index estimates Alewife reproductive success the preceding year. The warm summer and winter of 2016 likely contributed to the large year class. In contrast the relatively cool 2017 spring and cold winter may result in a lower than average 2017 year class. Abundance indices for Rainbow Smelt, Cisco, and Emerald Shiner either declined or remained at low levels in 2017. Pelagic prey fish diversity continues to be low since a single species, Alewife, dominates the catch. Deepwater Sculpin were the most abundant benthic prey fish in 2017 because Round Goby abundance declined sharply from 2016. Slimy Sculpin density continued to decline and the 2017 biomass index for US waters was the lowest ever observed. Prior to Round Goby proliferation, juvenile Slimy Sculpin comprised ~10% of the Slimy Sculpin catch, but

Common Fluorescence In Situ Hybridization Applications in Cytology.

PubMed

Savic, Spasenija; Bubendorf, Lukas

2016-12-01

- Fluorescence in situ hybridization (FISH) is a well-established method for detection of genomic aberrations in diagnostic, prognostic, and predictive marker testing. - To review common applications of FISH in cytology. - The published literature was reviewed. - Cytology is particularly well suited for all kinds of FISH applications, which is highlighted in respiratory tract cytology with an increasing demand for predictive FISH testing in lung cancer. Fluorescence in situ hybridization is the gold standard for detection of predictive anaplastic lymphoma kinase gene (ALK) rearrangements, and the same evaluation criteria as in histology apply to cytology. Several other gene rearrangements, including ROS proto-oncogene 1 receptor tyrosine kinase (ROS1), are becoming clinically important and share the same underlining cytogenetic mechanisms with ALK. MET amplification is one of the most common mechanisms of acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors and can be targeted by crizotinib. As genomic aberrations are a hallmark of malignant cells, FISH is a valuable objective ancillary diagnostic tool. In urinary tract cytology, atypical urothelial cells equivocal for malignancy are a common diagnostic dilemma and multitarget FISH can help clarify such cells. Diagnosis of malignant mesothelioma remains one of the most challenging fields in effusion cytology, and ancillary FISH is useful in establishing the diagnosis. Fluorescence in situ hybridization is a morphology-based technique, and the prerequisite for reliable FISH results is a targeted evaluation of the cells in question (eg, cancer or atypical cells). Cytopathologists and cytotechnicians should therefore be involved in molecular testing in order to select the best material and to provide their morphologic expertise.

FISH Finder: a high-throughput tool for analyzing FISH images

PubMed Central

Shirley, James W.; Ty, Sereyvathana; Takebayashi, Shin-ichiro; Liu, Xiuwen; Gilbert, David M.

2011-01-01

Motivation: Fluorescence in situ hybridization (FISH) is used to study the organization and the positioning of specific DNA sequences within the cell nucleus. Analyzing the data from FISH images is a tedious process that invokes an element of subjectivity. Automated FISH image analysis offers savings in time as well as gaining the benefit of objective data analysis. While several FISH image analysis software tools have been developed, they often use a threshold-based segmentation algorithm for nucleus segmentation. As fluorescence signal intensities can vary significantly from experiment to experiment, from cell to cell, and within a cell, threshold-based segmentation is inflexible and often insufficient for automatic image analysis, leading to additional manual segmentation and potential subjective bias. To overcome these problems, we developed a graphical software tool called FISH Finder to automatically analyze FISH images that vary significantly. By posing the nucleus segmentation as a classification problem, compound Bayesian classifier is employed so that contextual information is utilized, resulting in reliable classification and boundary extraction. This makes it possible to analyze FISH images efficiently and objectively without adjustment of input parameters. Additionally, FISH Finder was designed to analyze the distances between differentially stained FISH probes. Availability: FISH Finder is a standalone MATLAB application and platform independent software. The program is freely available from: http://code.google.com/p/fishfinder/downloads/list Contact: gilbert@bio.fsu.edu PMID:21310746

The campaign to DNA barcode all fishes, FISH-BOL.

PubMed

Ward, R D; Hanner, R; Hebert, P D N

2009-02-01

FISH-BOL, the Fish Barcode of Life campaign, is an international research collaboration that is assembling a standardized reference DNA sequence library for all fishes. Analysis is targeting a 648 base pair region of the mitochondrial cytochrome c oxidase I (COI) gene. More than 5000 species have already been DNA barcoded, with an average of five specimens per species, typically vouchers with authoritative identifications. The barcode sequence from any fish, fillet, fin, egg or larva can be matched against these reference sequences using BOLD; the Barcode of Life Data System (http://www.barcodinglife.org). The benefits of barcoding fishes include facilitating species identification, highlighting cases of range expansion for known species, flagging previously overlooked species and enabling identifications where traditional methods cannot be applied. Results thus far indicate that barcodes separate c. 98 and 93% of already described marine and freshwater fish species, respectively. Several specimens with divergent barcode sequences have been confirmed by integrative taxonomic analysis as new species. Past concerns in relation to the use of fish barcoding for species discrimination are discussed. These include hybridization, recent radiations, regional differentiation in barcode sequences and nuclear copies of the barcode region. However, current results indicate these issues are of little concern for the great majority of specimens.

Determination of the ruminant origin of bone particles using fluorescence in situ hybridization (FISH).

PubMed

Lecrenier, M C; Ledoux, Q; Berben, G; Fumière, O; Saegerman, C; Baeten, V; Veys, P

2014-07-17

Molecular biology techniques such as PCR constitute powerful tools for the determination of the taxonomic origin of bones. DNA degradation and contamination by exogenous DNA, however, jeopardise bone identification. Despite the vast array of techniques used to decontaminate bone fragments, the isolation and determination of bone DNA content are still problematic. Within the framework of the eradication of transmissible spongiform encephalopathies (including BSE, commonly known as "mad cow disease"), a fluorescence in situ hybridization (FISH) protocol was developed. Results from the described study showed that this method can be applied directly to bones without a demineralisation step and that it allows the identification of bovine and ruminant bones even after severe processing. The results also showed that the method is independent of exogenous contamination and that it is therefore entirely appropriate for this application.

Monosomy 3 by FISH in uveal melanoma: variability in techniques and results.

PubMed

Aronow, Mary; Sun, Yang; Saunthararajah, Yogen; Biscotti, Charles; Tubbs, Raymond; Triozzi, Pierre; Singh, Arun D

2012-09-01

Tumor monosomy 3 confers a poor prognosis in patients with uveal melanoma. We critically review the techniques used for fluorescence in situ hybridization (FISH) detection of monosomy 3 in order to assess variability in practice patterns and to explain differences in results. Significant variability that has likely affected reported results was found in tissue sampling methods, selection of FISH probes, number of cells counted, and the cut-off point used to determine monosomy 3 status. Clinical parameters and specific techniques employed to report FISH results should be specified so as to allow meta-analysis of published studies. FISH-based detection of monosomy 3 in uveal melanoma has not been performed in a standardized manner, which limits conclusions regarding its clinical utility. FISH is a widely available, versatile technology, and when performed optimally has the potential to be a valuable tool for determining the prognosis of uveal melanoma. Copyright © 2012 Elsevier Inc. All rights reserved.

Probabilistic framework for assessing the arsenic exposure risk from cooked fish consumption.

PubMed

Ling, Min-Pei; Wu, Chiu-Hua; Chen, Szu-Chieh; Chen, Wei-Yu; Chio, Chia-Pin; Cheng, Yi-Hsien; Liao, Chung-Min

2014-12-01

Geogenic arsenic (As) contamination of groundwater is a major ecological and human health problem in southwestern and northeastern coastal areas of Taiwan. Here, we present a probabilistic framework for assessing the human health risks from consuming raw and cooked fish that were cultured in groundwater As-contaminated ponds in Taiwan by linking a physiologically based pharmacokinetics model and a Weibull dose-response model. Results indicate that As levels in baked, fried, and grilled fish were higher than those of raw fish. Frying resulted in the greatest increase in As concentration, followed by grilling, with baking affecting the As concentration the least. Simulation results show that, following consumption of baked As-contaminated fish, the health risk to humans is <10(-6) excess bladder cancer risk level for lifetime exposure; as the incidence ratios of liver and lung cancers are generally acceptable at risk ranging from 10(-6) to 10(-4), the consumption of baked As-contaminated fish is unlikely to pose a significant risk to human health. However, contaminated fish cooked by frying resulted in significant health risks, showing the highest cumulative incidence ratios of liver cancer. We also show that males have higher cumulative incidence ratio of liver cancer than females. We found that although cooking resulted in an increase for As levels in As-contaminated fish, the risk to human health of consuming baked fish is nevertheless acceptable. We suggest the adoption of baking as a cooking method and warn against frying As-contaminated fish. We conclude that the concentration of contaminants after cooking should be taken into consideration when assessing the risk to human health.

Towards Integration of Environmental and Health Impact Assessments for Wild Capture Fishing and Farmed Fish with Particular Reference to Public Health and Occupational Health Dimensions

PubMed Central

Watterson, Andrew; Little, David; Young, James A.; Boyd, Kathleen; Azim, Ekram; Murray, Francis

2008-01-01

The paper offers a review and commentary, with particular reference to the production of fish from wild capture fisheries and aquaculture, on neglected aspects of health impact assessments which are viewed by a range of international and national health bodies and development agencies as valuable and necessary project tools. Assessments sometimes include environmental health impact assessments but rarely include specific occupational health and safety impact assessments especially integrated into a wider public health assessment. This is in contrast to the extensive application of environmental impact assessments to fishing and the comparatively large body of research now generated on the public health effects of eating fish. The value of expanding and applying the broader assessments would be considerable because in 2004 the United Nations Food and Agriculture Organization reports there were 41,408,000 people in the total ‘fishing’ sector including 11,289,000 in aquaculture. The paper explores some of the complex interactions that occur with regard to fishing activities and proposes the wider adoption of health impact assessment tools in these neglected sectors through an integrated public health impact assessment tool. PMID:19190356

Fish bioaccumulation and biomarkers in environmental risk assessment: a review.

PubMed

van der Oost, Ron; Beyer, Jonny; Vermeulen, Nico P E

2003-02-01

In this review, a wide array of bioaccumulation markers and biomarkers, used to demonstrate exposure to and effects of environmental contaminants, has been discussed in relation to their feasibility in environmental risk assessment (ERA). Fish bioaccumulation markers may be applied in order to elucidate the aquatic behavior of environmental contaminants, as bioconcentrators to identify certain substances with low water levels and to assess exposure of aquatic organisms. Since it is virtually impossible to predict the fate of xenobiotic substances with simple partitioning models, the complexity of bioaccumulation should be considered, including toxicokinetics, metabolism, biota-sediment accumulation factors (BSAFs), organ-specific bioaccumulation and bound residues. Since it remains hard to accurately predict bioaccumulation in fish, even with highly sophisticated models, analyses of tissue levels are required. The most promising fish bioaccumulation markers are body burdens of persistent organic pollutants, like PCBs and DDTs. Since PCDD and PCDF levels in fish tissues are very low as compared with the sediment levels, their value as bioaccumulation markers remains questionable. Easily biodegradable compounds, such as PAHs and chlorinated phenols, do not tend to accumulate in fish tissues in quantities that reflect the exposure. Semipermeable membrane devices (SPMDs) have been successfully used to mimic bioaccumulation of hydrophobic organic substances in aquatic organisms. In order to assess exposure to or effects of environmental pollutants on aquatic ecosystems, the following suite of fish biomarkers may be examined: biotransformation enzymes (phase I and II), oxidative stress parameters, biotransformation products, stress proteins, metallothioneins (MTs), MXR proteins, hematological parameters, immunological parameters, reproductive and endocrine parameters, genotoxic parameters, neuromuscular parameters, physiological, histological and morphological parameters

An assessment of fish assemblage structure in a large river

USGS Publications Warehouse

Kiraly, Ian A.; Coghlan, S.M.; Zydlewski, Joseph D.; Hayes, D.

2015-01-01

The Penobscot River drains the largest watershed in Maine and once provided spawning and rearing habitats to 11 species of diadromous fishes. The construction of dams blocked migrations of these fishes and likely changed the structure and function of fish assemblages throughout the river. The proposed removal of two main-stem dams, improved upstream fish passage at a third dam, and construction of a fish bypass on a dam obstructing a major tributary is anticipated to increase passage of and improve habitat connectivity for both diadromous and resident fishes. We captured 61 837 fish of 35 species in the Penobscot River and major tributaries, through 114 km of boat electrofishing. Patterns of fish assemblage structure did not change considerably during our sampling; relatively few species contributed to seasonal and annual variability within the main-stem river, including smallmouth bass Micropterus dolomieu, white sucker Catostomus commersonii, pumpkinseed Lepomis gibbosus, and golden shiner Notemigonus crysoleucas. However, distinct fish assemblages were present among river sections bounded by dams. Many diadromous species were restricted to tidal waters downriver of the Veazie Dam; Fundulus species were also abundant within the tidal river section. Smallmouth bass and pumpkinseed were most prevalent within the Veazie Dam impoundment and the free-flowing river section immediately upriver, suggesting the importance of both types of habitat that supports multiple life stages of these species. Further upriver, brown bullhead Ameiurus nebulosus, yellow perch Perca flavescens, chain pickerel Esox niger, and cyprinid species were more prevalent than within any other river section. Our findings describe baseline spatial patterns of fish assemblages in the Penobscot River in relation to dams with which to compare assessments after dam removal occurs.

[Development of an original computer program FISHMet: use for molecular cytogenetic diagnosis and genome mapping by fluorescent in situ hybridization (FISH)].

PubMed

Iurov, Iu B; Khazatskiĭ, I A; Akindinov, V A; Dovgilov, L V; Kobrinskiĭ, B A; Vorsanova, S G

2000-08-01

Original software FISHMet has been developed and tried for improving the efficiency of diagnosis of hereditary diseases caused by chromosome aberrations and for chromosome mapping by fluorescent in situ hybridization (FISH) method. The program allows creation and analysis of pseudocolor chromosome images and hybridization signals in the Windows 95 system, allows computer analysis and editing of the results of pseudocolor hybridization in situ, including successive imposition of initial black-and-white images created using fluorescent filters (blue, green, and red), and editing of each image individually or of a summary pseudocolor image in BMP, TIFF, and JPEG formats. Components of image computer analysis system (LOMO, Leitz Ortoplan, and Axioplan fluorescent microscopes, COHU 4910 and Sanyo VCB-3512P CCD cameras, Miro-Video, Scion LG-3 and VG-5 image capture maps, and Pentium 100 and Pentium 200 computers) and specialized software for image capture and visualization (Scion Image PC and Video-Cup) have been used with good results in the study.

Large-scale assessment of Mediterranean marine protected areas effects on fish assemblages.

PubMed

Guidetti, Paolo; Baiata, Pasquale; Ballesteros, Enric; Di Franco, Antonio; Hereu, Bernat; Macpherson, Enrique; Micheli, Fiorenza; Pais, Antonio; Panzalis, Pieraugusto; Rosenberg, Andrew A; Zabala, Mikel; Sala, Enric

2014-01-01

Marine protected areas (MPAs) were acknowledged globally as effective tools to mitigate the threats to oceans caused by fishing. Several studies assessed the effectiveness of individual MPAs in protecting fish assemblages, but regional assessments of multiple MPAs are scarce. Moreover, empirical evidence on the role of MPAs in contrasting the propagation of non-indigenous-species (NIS) and thermophilic species (ThS) is missing. We simultaneously investigated here the role of MPAs in reversing the effects of overfishing and in limiting the spread of NIS and ThS. The Mediterranean Sea was selected as study area as it is a region where 1) MPAs are numerous, 2) fishing has affected species and ecosystems, and 3) the arrival of NIS and the northward expansion of ThS took place. Fish surveys were done in well-enforced no-take MPAs (HP), partially-protected MPAs (IP) and fished areas (F) at 30 locations across the Mediterranean. Significantly higher fish biomass was found in HP compared to IP MPAs and F. Along a recovery trajectory from F to HP MPAs, IP were similar to F, showing that just well enforced MPAs triggers an effective recovery. Within HP MPAs, trophic structure of fish assemblages resembled a top-heavy biomass pyramid. Although the functional structure of fish assemblages was consistent among HP MPAs, species driving the recovery in HP MPAs differed among locations: this suggests that the recovery trajectories in HP MPAs are likely to be functionally similar (i.e., represented by predictable changes in trophic groups, especially fish predators), but the specific composition of the resulting assemblages may depend on local conditions. Our study did not show any effect of MPAs on NIS and ThS. These results may help provide more robust expectations, at proper regional scale, about the effects of new MPAs that may be established in the Mediterranean Sea and other ecoregions worldwide.

Responses of hybrid striped bass to waterborne and dietary copper in freshwater and saltwater

USGS Publications Warehouse

Bielmyer, G.K.; Gatlin, D.; Isely, J.J.; Tomasso, J.; Klaine, S.J.

2005-01-01

Mechanisms of copper toxicity and consequences of exposure vary due to uptake route and ionoregulatory status. The goal of this research was to develop a model fish system to assess the influence of different Cu exposure routes (waterborne or dietary) on bioavailability, uptake, and effects in hybrid striped bass (Morone chrysops×Morone saxatilis) acclimated to fresh- or saltwater. Initially, hybrid striped bass were exposed to dietary Cu concentrations of 571, 785, and 1013 μg Cu/g, along with a control (∼ 5 μg Cu/g), for 14 days in saltwater. Intestinal and liver Cu accumulated in a dose-dependent manner in fish exposed to increasing levels of dietary Cu. Chronic (42 days) experiments were then conducted to determine sub-lethal effects of aqueous, dietary, and combined aqueous and dietary Cu exposures to both freshwater- and saltwater-acclimated hybrid striped bass. Growth and Cu accumulation in the gill, intestine, and liver were measured. Although no significant effects were observed in fish exposed to waterborne Cu, those exposed through the diet accumulated significant liver and intestinal Cu but showed no significant change in growth. Overall, these results suggest that at the levels tested, exposure to elevated waterborne Cu did not cause significant long-term tissue Cu accumulation, whereas dietary Cu exposure caused significant liver and intestinal Cu accumulation in hybrid striped bass which was comparable in both freshwater and saltwater (15 g/L).

Optimized RNA ISH, RNA FISH and protein-RNA double labeling (IF/FISH) in Drosophila ovaries

PubMed Central

Zimmerman, Sandra G; Peters, Nathaniel C; Altaras, Ariel E; Berg, Celeste A

2014-01-01

In situ hybridization (ISH) is a powerful technique for detecting nucleic acids in cells and tissues. Here we describe three ISH procedures that are optimized for Drosophila ovaries: whole-mount, digoxigenin-labeled RNA ISH; RNA fluorescent ISH (FISH); and protein immunofluorescence (IF)–RNA FISH double labeling (IF/FISH). Each procedure balances conflicting requirements for permeabilization, fixation and preservation of antigenicity to detect RNA and protein expression with high resolution and sensitivity. The ISH protocol uses alkaline phosphatase–conjugated digoxigenin antibodies followed by a color reaction, whereas FISH detection involves tyramide signal amplification (TSA). To simultaneously preserve antigens for protein detection and enable RNA probe penetration for IF/FISH, we perform IF before FISH and use xylenes and detergents to permeabilize the tissue rather than proteinase K, which can damage the antigens. ISH and FISH take 3 d to perform, whereas IF/FISH takes 5 d. Probe generation takes 1 or 2 d to perform. PMID:24113787

SURROGATE SPECIES IN ASSESSING CONTAMINANT RISK FOR ENDANGERED FISHES

EPA Science Inventory

Rainbow trout, fathead minnows, and sheepshead minnows were tested as surrogate species to assess contaminant risk for 17 endangered fishes and one toad species. Acute toxicity tests were conducted with carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin in accord...

Threshold-dependent sample sizes for selenium assessment with stream fish tissue

USGS Publications Warehouse

Hitt, Nathaniel P.; Smith, David R.

2015-01-01

Natural resource managers are developing assessments of selenium (Se) contamination in freshwater ecosystems based on fish tissue concentrations. We evaluated the effects of sample size (i.e., number of fish per site) on the probability of correctly detecting mean whole-body Se values above a range of potential management thresholds. We modeled Se concentrations as gamma distributions with shape and scale parameters fitting an empirical mean-to-variance relationship in data from southwestern West Virginia, USA (63 collections, 382 individuals). We used parametric bootstrapping techniques to calculate statistical power as the probability of detecting true mean concentrations up to 3 mg Se/kg above management thresholds ranging from 4 to 8 mg Se/kg. Sample sizes required to achieve 80% power varied as a function of management thresholds and Type I error tolerance (α). Higher thresholds required more samples than lower thresholds because populations were more heterogeneous at higher mean Se levels. For instance, to assess a management threshold of 4 mg Se/kg, a sample of eight fish could detect an increase of approximately 1 mg Se/kg with 80% power (given α = 0.05), but this sample size would be unable to detect such an increase from a management threshold of 8 mg Se/kg with more than a coin-flip probability. Increasing α decreased sample size requirements to detect above-threshold mean Se concentrations with 80% power. For instance, at an α-level of 0.05, an 8-fish sample could detect an increase of approximately 2 units above a threshold of 8 mg Se/kg with 80% power, but when α was relaxed to 0.2, this sample size was more sensitive to increasing mean Se concentrations, allowing detection of an increase of approximately 1.2 units with equivalent power. Combining individuals into 2- and 4-fish composite samples for laboratory analysis did not decrease power because the reduced number of laboratory samples was compensated for by increased

Determination of the ruminant origin of bone particles using fluorescence in situ hybridization (FISH)

PubMed Central

Lecrenier, M. C.; Ledoux, Q.; Berben, G.; Fumière, O.; Saegerman, C.; Baeten, V.; Veys, P.

2014-01-01

Molecular biology techniques such as PCR constitute powerful tools for the determination of the taxonomic origin of bones. DNA degradation and contamination by exogenous DNA, however, jeopardise bone identification. Despite the vast array of techniques used to decontaminate bone fragments, the isolation and determination of bone DNA content are still problematic. Within the framework of the eradication of transmissible spongiform encephalopathies (including BSE, commonly known as “mad cow disease”), a fluorescence in situ hybridization (FISH) protocol was developed. Results from the described study showed that this method can be applied directly to bones without a demineralisation step and that it allows the identification of bovine and ruminant bones even after severe processing. The results also showed that the method is independent of exogenous contamination and that it is therefore entirely appropriate for this application. PMID:25034259

Fish Passage Assessment: Big Canyon Creek Watershed, Technical Report 2004.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Christian, Richard

2004-02-01

This report presents the results of the fish passage assessment as outlined as part of the Protect and Restore the Big Canyon Creek Watershed project as detailed in the CY2003 Statement of Work (SOW). As part of the Northwest Power Planning Council's Columbia Basin Fish and Wildlife Program (FWP), this project is one of Bonneville Power Administration's (BPA) many efforts at off-site mitigation for damage to salmon and steelhead runs, their migration, and wildlife habitat caused by the construction and operation of federal hydroelectric dams on the Columbia River and its tributaries. The proposed restoration activities within the Big Canyonmore » Creek watershed follow the watershed restoration approach mandated by the Fisheries and Watershed Program. Nez Perce Tribal Fisheries/Watershed Program vision focuses on protecting, restoring, and enhancing watersheds and treaty resources within the ceded territory of the Nez Perce Tribe under the Treaty of 1855 with the United States Federal Government. The program uses a holistic approach, which encompasses entire watersheds, ridge top to ridge top, emphasizing all cultural aspects. We strive toward maximizing historic ecosystem productive health, for the restoration of anadromous and resident fish populations. The Nez Perce Tribal Fisheries/Watershed Program (NPTFWP) sponsors the Protect and Restore the Big Canyon Creek Watershed project. The NPTFWP has the authority to allocate funds under the provisions set forth in their contract with BPA. In the state of Idaho vast numbers of relatively small obstructions, such as road culverts, block thousands of miles of habitat suitable for a variety of fish species. To date, most agencies and land managers have not had sufficient, quantifiable data to adequately address these barrier sites. The ultimate objective of this comprehensive inventory and assessment was to identify all barrier crossings within the watershed. The barriers were then prioritized according to the amount

FISH-in-CHIPS: A Microfluidic Platform for Molecular Typing of Cancer Cells.

PubMed

Perez-Toralla, Karla; Mottet, Guillaume; Tulukcuoglu-Guneri, Ezgi; Champ, Jérôme; Bidard, François-Clément; Pierga, Jean-Yves; Klijanienko, Jerzy; Draskovic, Irena; Malaquin, Laurent; Viovy, Jean-Louis; Descroix, Stéphanie

2017-01-01

Microfluidics offer powerful tools for the control, manipulation, and analysis of cells, in particular for the assessment of cell malignancy or the study of cell subpopulations. However, implementing complex biological protocols on chip remains a challenge. Sample preparation is often performed off chip using multiple manually performed steps, and protocols usually include different dehydration and drying steps that are not always compatible with a microfluidic format.Here, we report the implementation of a Fluorescence in situ Hybridization (FISH) protocol for the molecular typing of cancer cells in a simple and low-cost device. The geometry of the chip allows integrating the sample preparation steps to efficiently assess the genomic content of individual cells using a minute amount of sample. The FISH protocol can be fully automated, thus enabling its use in routine clinical practice.

Correlation of IHC and FISH for ALK gene rearrangement in non-small cell lung carcinoma: IHC score algorithm for FISH.

PubMed

Yi, Eunhee S; Boland, Jennifer M; Maleszewski, Joseph J; Roden, Anja C; Oliveira, Andre M; Aubry, Marie-Christine; Erickson-Johnson, Michele R; Caron, Bolette L; Li, Yan; Tang, Hui; Stoddard, Shawn; Wampfler, Jason; Kulig, Kimary; Yang, Ping

2011-03-01

Accurate, cost-effective methods for testing anaplastic lymphoma kinase gene rearrangement (ALK+) are needed to select patients with non-small cell lung carcinoma for ALK-inhibitor therapy. Fluorescent in situ hybridization (FISH) is used to detect ALK+, but it is expensive and not routinely available. We explored the potential of an immunohistochemistry (IHC) scoring system as an affordable, accessible approach. One hundred one samples were obtained from an enriched cohort of never-smokers with adenocarcinoma from the Mayo Clinic Lung Cancer Cohort. IHC was performed using the ALK1 monoclonal antibody with ADVANCE detection system (Dako) and FISH with dual-color, break-apart probe (Abbott Molecular) on formalin-fixed, paraffin-embedded tissue. Cases were assessed as IHC score 0 (no staining; n = 69), 1+ (faint cytoplasmic staining, n = 21), 2+ (moderate, smooth cytoplasmic staining; n = 3), or 3+ (intense, granular cytoplasmic staining in ≥10% of tumor cells; n = 8). All IHC 3+ cases were FISH+, whereas 1 of 3 IHC 2+ and 1 of 21 IHC 1+ cases were FISH+. All 69 IHC 0 cases were FISH-. Considering FISH a gold-standard reference in this study, sensitivity and specificity of IHC were 90 and 97.8%, respectively, when 2+ and 3+ were regarded as IHC positive and 0 and 1+ as IHC negative. IHC scoring correlates with FISH and may be a useful algorithm in testing ALK+ by FISH in non-small cell lung carcinoma, similar to human epidermal growth factor-2 testing in breast cancer. Further study is needed to validate this approach.

Assessing the natural and anthropogenic influences on basin-wide fish species richness.

PubMed

Cheng, Su-Ting; Herricks, Edwin E; Tsai, Wen-Ping; Chang, Fi-John

2016-12-01

Theory predicts that the number of fish species increases with river size in natural free-flowing rivers, but the relationship is lost under intensive exploitation of water resources associated with dams and/or landscape developments. In this paper, we aim to identify orthomorphic issues that disrupt theoretical species patterns based on a multi-year, basin-wide assessment in the Danshuei River Watershed of Taiwan. We hypothesize that multiple human-induced modifications fragment habitat areas leading to decreases of local fish species richness. We integrally relate natural and anthropogenic influences on fish species richness by a multiple linear regression model that is driven by a combination of factors including river network structure controls, water quality alterations of habitat, and disruption of channel connectivity with major discontinuities in habitat caused by dams. We found that stream order is a major forcing factor representing natural influence on fish species richness. In addition to stream order, we identified dams, dissolved oxygen deficiency (DO), and excessive total phosphorus (TP) as major anthropogenic influences on the richness of fish species. Our results showed that anthropogenic influences were operating at various spatial scales that inherently regulate the physical, chemical, and biological condition of fish habitats. Moreover, our probability-based risk assessment revealed causes of species richness reduction and opportunities for mitigation. Risks of species richness reduction caused by dams were determined by the position of dams and the contribution of tributaries in the drainage network. Risks associated with TP and DO were higher in human-activity-intensified downstream reaches. Our methodology provides a structural framework for assessing changes in basin-wide fish species richness under the mixed natural and human-modified river network and habitat conditions. Based on our analysis results, we recommend that a focus on landscape

A protocol for identifying suitable biomarkers to assess fish health: A systematic review

PubMed Central

2017-01-01

Background Biomarkers have been used extensively to provide the connection between external levels of contaminant exposure, internal levels of tissue contamination, and early adverse effects in organisms. Objectives To present a three-step protocol for identifying suitable biomarkers to assess fish health in coastal and marine ecosystems, using Gladstone Harbour (Australia) as a case study. Methods Prior to applying our protocol, clear working definitions for biomarkers were developed to ensure consistency with the global literature on fish health assessment. First, contaminants of concern were identified based on the presence of point and diffuse sources of pollution and available monitoring data for the ecosystem of interest. Second, suitable fish species were identified using fisheries dependent and independent data, and prioritised based on potential pathways of exposure to the contaminants of concern. Finally, a systematic and critical literature review was conducted on the use of biomarkers to assess the health of fish exposed to the contaminants of concern. Results/Discussion We present clear working definitions for bioaccumulation markers, biomarkers of exposure, biomarkers of effect and biomarkers of susceptibility. Based on emission and concentration information, seven metals were identified as contaminants of concern for Gladstone Harbour. Twenty out of 232 fish species were abundant enough to be potentially suitable for biomarker studies; five of these were prioritised based on potential pathways of exposure and susceptibility to metals. The literature search on biomarkers yielded 5,035 articles, of which 151met the inclusion criteria. Based on our review, the most suitable biomarkers include bioaccumulation markers, biomarkers of exposure (CYP1A, EROD, SOD, LPOX, HSP, MT, DNA strand breaks, micronuclei, apoptosis), and biomarkers of effect (histopathology, TAG:ST). Conclusion Our protocol outlines a clear pathway to identify suitable biomarkers to

Probabilistic assessment of uncertain adaptive hybrid composites

NASA Technical Reports Server (NTRS)

Shiao, Michael C.; Singhal, Surendra N.; Chamis, Christos C.

1994-01-01

Adaptive composite structures using actuation materials, such as piezoelectric fibers, were assessed probabilistically utilizing intraply hybrid composite mechanics in conjunction with probabilistic composite structural analysis. Uncertainties associated with the actuation material as well as the uncertainties in the regular (traditional) composite material properties were quantified and considered in the assessment. Static and buckling analyses were performed for rectangular panels with various boundary conditions and different control arrangements. The probability density functions of the structural behavior, such as maximum displacement and critical buckling load, were computationally simulated. The results of the assessment indicate that improved design and reliability can be achieved with actuation material.

A CO-FISH assay to assess sister chromatid segregation patterns in mitosis of mouse embryonic stem cells.

PubMed

Sauer, Stephan; Burkett, Sandra S; Lewandoski, Mark; Klar, Amar J S

2013-05-01

Sister chromatids contain identical DNA sequence but are chiral with respect to both their helical handedness and their replication history. Emerging evidence from various model organisms suggests that certain stem cells segregate sister chromatids nonrandomly to either maintain genome integrity or to bias cellular differentiation in asymmetric cell divisions. Conventional methods for tracing of old vs. newly synthesized DNA strands generally lack resolution for individual chromosomes and employ halogenated thymidine analogs with profound cytotoxic effects on rapidly dividing cells. Here, we present a modified chromosome orientation fluorescence in situ hybridization (CO-FISH) assay, where identification of individual chromosomes and their replication history is achieved in subsequent hybridization steps with chromosome-specific DNA probes and PNA telomere probes. Importantly, we tackle the issue of BrdU cytotoxicity and show that our method is compatible with normal mouse ES cell biology, unlike a recently published related protocol. Results from our CO-FISH assay show that mitotic segregation of mouse chromosome 7 is random in ES cells, which contrasts previously published results from our laboratory and settles a controversy. Our straightforward protocol represents a useful resource for future studies on chromatid segregation patterns of in vitro-cultured cells from distinct model organisms.

Best practices for assessing forage fish fisheries-seabird resource competition

USGS Publications Warehouse

Sydeman, William J.; Thompson, Sarah Ann; Anker-Nilssen, Tycho; Arimitsu, Mayumi L.; Bennison, Ashley; Bertrand, Sophie; Boersch-Supan, Philipp; Boyd, Charlotte; Bransome, Nicole C.; Crawford, Robert J.M.; Daunt, Francis; Furness, Robert W.; Gianuca, Dimas; Gladics, Amanda; Koehn, Laura; Lang, Jennifer W.; Loggerwell, Elizabeth; Morris, Taryn L.; Phillips, Elizabeth M.; Provencher, Jennifer; Punt, André E..; Saraux, Claire; Shannon, Lynne; Sherley, Richard B.; Simeone, Alejandro; Wanless, Ross M.; Wanless, Sarah; Zador, Stephani

2017-01-01

Worldwide, in recent years capture fisheries targeting lower-trophic level forage fish and euphausiid crustaceans have been substantial (∼20 million metric tons [MT] annually). Landings of forage species are projected to increase in the future, and this harvest may affect marine ecosystems and predator-prey interactions by removal or redistribution of biomass central to pelagic food webs. In particular, fisheries targeting forage fish and euphausiids may be in competition with seabirds, likely the most sensitive of marine vertebrates given limitations in their foraging abilities (ambit and gape size) and high metabolic rate, for food resources. Lately, apparent competition between fisheries and seabirds has led to numerous high-profile conflicts over interpretations, as well as the approaches that could and should be used to assess the magnitude and consequences of fisheries-seabird resource competition. In this paper, we review the methods used to date to study fisheries competition with seabirds, and present “best practices” for future resource competition assessments. Documenting current fisheries competition with seabirds generally involves addressing two major issues: 1) are fisheries causing localized prey depletion that is sufficient to affect the birds? (i.e., are fisheries limiting food resources?), and 2) how are fisheries-induced changes to forage stocks affecting seabird populations given the associated functional or numerical response relationships? Previous studies have been hampered by mismatches in the scale of fisheries, fish, and seabird data, and a lack of causal understanding due to confounding by climatic and other ecosystem factors (e.g., removal of predatory fish). Best practices for fisheries-seabird competition research should include i) clear articulation of hypotheses, ii) data collection (or summation) of fisheries, fish, and seabirds on matched spatio-temporal scales, and iii) integration of observational and experimental

Fish Vaccine Development and Use to Prevent Streptococcal Diseases

USDA-ARS?s Scientific Manuscript database

An important pathogen of tilapia, hybrid striped bass and trout raised in intensive aquaculture is Streptococcus sp., a cause of severe economic losses in the fish farming industry. Infected fish experience severe to moderate mortality due to Streptococcus iniae and/or S. agalactiae. The diseased ...

Large-Scale Assessment of Mediterranean Marine Protected Areas Effects on Fish Assemblages

PubMed Central

Guidetti, Paolo; Baiata, Pasquale; Ballesteros, Enric; Di Franco, Antonio; Hereu, Bernat; Macpherson, Enrique; Micheli, Fiorenza; Pais, Antonio; Panzalis, Pieraugusto; Rosenberg, Andrew A.; Zabala, Mikel; Sala, Enric

2014-01-01

Marine protected areas (MPAs) were acknowledged globally as effective tools to mitigate the threats to oceans caused by fishing. Several studies assessed the effectiveness of individual MPAs in protecting fish assemblages, but regional assessments of multiple MPAs are scarce. Moreover, empirical evidence on the role of MPAs in contrasting the propagation of non-indigenous-species (NIS) and thermophilic species (ThS) is missing. We simultaneously investigated here the role of MPAs in reversing the effects of overfishing and in limiting the spread of NIS and ThS. The Mediterranean Sea was selected as study area as it is a region where 1) MPAs are numerous, 2) fishing has affected species and ecosystems, and 3) the arrival of NIS and the northward expansion of ThS took place. Fish surveys were done in well-enforced no-take MPAs (HP), partially-protected MPAs (IP) and fished areas (F) at 30 locations across the Mediterranean. Significantly higher fish biomass was found in HP compared to IP MPAs and F. Along a recovery trajectory from F to HP MPAs, IP were similar to F, showing that just well enforced MPAs triggers an effective recovery. Within HP MPAs, trophic structure of fish assemblages resembled a top-heavy biomass pyramid. Although the functional structure of fish assemblages was consistent among HP MPAs, species driving the recovery in HP MPAs differed among locations: this suggests that the recovery trajectories in HP MPAs are likely to be functionally similar (i.e., represented by predictable changes in trophic groups, especially fish predators), but the specific composition of the resulting assemblages may depend on local conditions. Our study did not show any effect of MPAs on NIS and ThS. These results may help provide more robust expectations, at proper regional scale, about the effects of new MPAs that may be established in the Mediterranean Sea and other ecoregions worldwide. PMID:24740479

Capturing ecology in modeling approaches applied to environmental risk assessment of endocrine active chemicals in fish.

PubMed

Mintram, Kate S; Brown, A Ross; Maynard, Samuel K; Thorbek, Pernille; Tyler, Charles R

2018-02-01

Endocrine active chemicals (EACs) are widespread in freshwater environments and both laboratory and field based studies have shown reproductive effects in fish at environmentally relevant exposures. Environmental risk assessment (ERA) seeks to protect wildlife populations and prospective assessments rely on extrapolation from individual-level effects established for laboratory fish species to populations of wild fish using arbitrary safety factors. Population susceptibility to chemical effects, however, depends on exposure risk, physiological susceptibility, and population resilience, each of which can differ widely between fish species. Population models have significant potential to address these shortfalls and to include individual variability relating to life-history traits, demographic and density-dependent vital rates, and behaviors which arise from inter-organism and organism-environment interactions. Confidence in population models has recently resulted in the EU Commission stating that results derived from reliable models may be considered when assessing the relevance of adverse effects of EACs at the population level. This review critically assesses the potential risks posed by EACs for fish populations, considers the ecological factors influencing these risks and explores the benefits and challenges of applying population modeling (including individual-based modeling) in ERA for EACs in fish. We conclude that population modeling offers a way forward for incorporating greater environmental relevance in assessing the risks of EACs for fishes and for identifying key risk factors through sensitivity analysis. Individual-based models (IBMs) allow for the incorporation of physiological and behavioral endpoints relevant to EAC exposure effects, thus capturing both direct and indirect population-level effects.

Using industry ROV videos to assess fish associations with subsea pipelines

NASA Astrophysics Data System (ADS)

McLean, D. L.; Partridge, J. C.; Bond, T.; Birt, M. J.; Bornt, K. R.; Langlois, T. J.

2017-06-01

Remote Operated Vehicles are routinely used to undertake inspection and maintenance activities of underwater pipelines in north-west Australia. In doing so, many terabytes of geo-referenced underwater video are collected at depths, and on a scale usually unobtainable for ecological research. We assessed fish diversity and abundance from existing ROV videos collected along 2-3 km sections of two pipelines in north-west Australia, one at 60-80 m water depth and the other at 120-130 m. A total of 5962 individual fish from 92 species and 42 families were observed. Both pipelines were characterised by a high abundance of commercially important fishes including: snappers (Lutjanidae) and groupers (Epinephelidae). The presence of thousands of unidentifiable larval fish, in addition to juveniles, sub-adults and adults suggests that the pipelines may be enhancing, rather than simply attracting, fish stocks. The prevalence and high complexity of sponges on the shallower pipeline and of deepwater corals on the deeper pipeline had a strong positive correlation with the fish abundance. These habitats likely offer a significant food source and refuge for fish, but also for invertebrates upon which fish feed. A greater diversity on the shallower pipeline, and a higher abundance of fishes on both pipelines, were associated with unsupported pipeline sections (spans) and many species appeared to be utilising pipeline spans as refuges. This study is a first look at the potential value of subsea pipelines for fishes on the north-west shelf. While the results suggest that these sections of pipeline appear to offer significant habitat that supports diverse and important commercially fished species, further work, including off-pipeline surveys on the natural seafloor, are required to determine conclusively the ecological value of pipelines and thereby inform discussions regarding the ecological implications of pipeline decommissioning.

The use of tumors in wild populations of fish to assess ecosystem health

USGS Publications Warehouse

Baumann, Paul C.

1992-01-01

Evidence has linked toxicants in aquatic systems with cancer in fish and population level effects on species. Thus some types of tumors may be useful monitors of ecosystem health, at least as affected by genotoxins and promoters. However, tumors caused by purely genetic mechanisms or by virus would not be good indicators. Only neoplasms which have chemicals as a portion of their etiology (either as initiators or promoters) would be useful in assessing ecosystem health. Lesions which may fit these criteria include liver neoplasms (both biliary and hepatic) and skin lesions in a variety of primarily benthic fishes, and neural lesions in various drum species and in butterfly fish species. Two studies purporting to demonstrate a lack of tumors in fish from polluted areas have been reexamined and found either to have insufficient data on vulnerable species or to actually support a tumor-pollution linkage. Thus certain lesions in vulnerable species or species groups may serve as a mechanism to assess one facet of ecosystem health.

Exposure assessment for trace elements from consumption of marine fish in Southeast Asia.

PubMed

Agusa, Tetsuro; Kunito, Takashi; Sudaryanto, Agus; Monirith, In; Kan-Atireklap, Supawat; Iwata, Hisato; Ismail, Ahmad; Sanguansin, Joompol; Muchtar, Muswerry; Tana, Touch Seang; Tanabe, Shinsuke

2007-02-01

Concentrations of 20 trace elements were determined in muscle and liver of 34 species of marine fish collected from coastal areas of Cambodia, Indonesia, Malaysia and Thailand. Large regional difference was observed in the levels of trace elements in liver of one fish family (Carangidae): the highest mean concentration was observed in fish from the Malaysian coastal waters for V, Cr, Zn, Pb and Bi and those from the Java Sea side of Indonesia for Sn and Hg. To assess the health risk to the Southeast Asian populations from consumption of fish, intake rates of trace elements were estimated. Some marine fish showed Hg levels higher than the guideline values by U.S. Environmental Protection Agency and Joint FAO/WHO Expert Committee on Food Additives (JECFA). This suggests that consumption of these fish may be hazardous to the people.

Expanding probe repertoire and improving reproducibility in human genomic hybridization

PubMed Central

Dorman, Stephanie N.; Shirley, Ben C.; Knoll, Joan H. M.; Rogan, Peter K.

2013-01-01

Diagnostic DNA hybridization relies on probes composed of single copy (sc) genomic sequences. Sc sequences in probe design ensure high specificity and avoid cross-hybridization to other regions of the genome, which could lead to ambiguous results that are difficult to interpret. We examine how the distribution and composition of repetitive sequences in the genome affects sc probe performance. A divide and conquer algorithm was implemented to design sc probes. With this approach, sc probes can include divergent repetitive elements, which hybridize to unique genomic targets under higher stringency experimental conditions. Genome-wide custom probe sets were created for fluorescent in situ hybridization (FISH) and microarray genomic hybridization. The scFISH probes were developed for detection of copy number changes within small tumour suppressor genes and oncogenes. The microarrays demonstrated increased reproducibility by eliminating cross-hybridization to repetitive sequences adjacent to probe targets. The genome-wide microarrays exhibited lower median coefficients of variation (17.8%) for two HapMap family trios. The coefficients of variations of commercial probes within 300 nt of a repetitive element were 48.3% higher than the nearest custom probe. Furthermore, the custom microarray called a chromosome 15q11.2q13 deletion more consistently. This method for sc probe design increases probe coverage for FISH and lowers variability in genomic microarrays. PMID:23376933

Influence of mercury bioaccessibility on exposure assessment associated with consumption of cooked predatory fish in Spain.

PubMed

Torres-Escribano, Silvia; Ruiz, Antonio; Barrios, Laura; Vélez, Dinoraz; Montoro, Rosa

2011-04-01

Predatory fish tend to accumulate high levels of mercury (Hg). Food safety assessment of these fish has been carried out on the raw product. However, the evaluation of the risk from Hg concentrations in raw fish might be modified if cooking and bioaccessibility (the contaminant fraction that solubilises from its matrix during gastrointestinal digestion and becomes available for intestinal absorption) were taken into account. Data on Hg bioaccessibility in raw predatory fish sold in Spain are scarce and no research on Hg bioaccessibility in cooked fish is available. The aim of the present study was to evaluate Hg bioaccessibility in various kinds of cooked predatory fish sold in Spain to estimate their health risk. Both Hg and bioaccessible Hg concentrations were analysed in raw and cooked fish (swordfish, tope shark, bonito and tuna). There were no changes in Hg concentrations during cooking. However, Hg bioaccessibility decreased significantly after cooking (42 ± 26% in raw fish and 26 ± 16% in cooked fish), thus reducing in swordfish and tope shark the Hg concentration to which the human organism would be exposed. In future, cooking and bioaccessibility should be considered in risk assessment of Hg concentrations in predatory fish. Copyright © 2011 Society of Chemical Industry.

Hybrid vehicle assessment. Phase 1: Petroleum savings analysis

NASA Technical Reports Server (NTRS)

Levin, R.; Liddle, S.; Deshpande, G.; Trummel, M.; Vivian, H. C.

1984-01-01

The results of a comprehensive analysis of near term electric hybrid vehicles are presented, with emphasis on their potential to save significant amounts of petroleum on a national scale in the 1990s. Performance requirements and expected annual usage patterns of these vehicles are first modeled. The projected U.S. fleet composition is estimated, and conceptual hybrid vehicle designs are conceived and analyzed for petroleum use when driven in the expected annual patterns. These petroleum consumption estimates are then compared to similar estimates for projected 1990 conventional vehicles having the same performance and driven in the same patterns. Results are presented in the form of three utility functions and comparisons of sevral conceptual designs are made. The Hybrid Vehicle (HV) design and assessment techniques are discussed and a general method is explained for selecting the optimum energy management strategy for any vehicle mission battery combination. Conclusions and recommendations are presented, and development recommendations are identified.

Identification of Cannabis sativa L. using the 1-kbTHCA synthase-fluorescence in situ hybridization probe.

PubMed

Jeangkhwoa, Pattraporn; Bandhaya, Achirapa; Umpunjun, Puangpaka; Chuenboonngarm, Ngarmnij; Panvisavas, Nathinee

2017-03-01

This study reports a successful application of fluorescence in situ hybridization (FISH) technique in the identification of Cannabis sativa L. cells recovered from fresh and dried powdered plant materials. Two biotin-16-dUTP-labeled FISH probes were designed from the Cannabis-specific tetrahydrocannabinolic acid synthase (THCAS) gene and the ITS region of the 45S rRNA gene. Specificity of probe-target hybridization was tested against the target and 4 non-target plant species, i.e., Humulus lupulus, Mitragyna speciosa, Papaver sp., and Nicotiana tabacum. The 1-kb THCA synthase hybridization probe gave Cannabis-specific hybridization signals, unlike the 700-bp Cannabis-ITS hybridization probe. Probe-target hybridization was also confirmed against 20 individual Cannabis plant samples. The 1-kb THCA synthase and 700-bp Cannabis-ITS hybridization probes clearly showed 2 hybridization signals per cell with reproducibility. The 1-kb THCA synthase probe did not give any FISH signal when tested against H. lupulus, its closely related member of the Canabaceae family. It was also showed that 1-kb THCA synthase FISH probe can be applied to identify small amount of dried powdered Cannabis material with an addition of rehydration step prior to the experimental process. This study provided an alternative identification method for Cannabis trace. Copyright © 2016. Published by Elsevier B.V.

Assessing the efficacy of single-pass backpack electrofishing to characterize fish community structure

USGS Publications Warehouse

Meador, M.R.; McIntyre, J.P.; Pollock, K.H.

2003-01-01

Two-pass backpack electrofishing data collected as part of the U.S. Geological Survey's National Water-Quality Assessment Program were analyzed to assess the efficacy of single-pass backpack electrofishing. A two-capture removal model was used to estimate, within 10 river basins across the United States, proportional fish species richness from one-pass electrofishing and probabilities of detection for individual fish species. Mean estimated species richness from first-pass sampling (ps1) ranged from 80.7% to 100% of estimated total species richness for each river basin, based on at least seven samples per basin. However, ps1 values for individual sites ranged from 40% to 100% of estimated total species richness. Additional species unique to the second pass were collected in 50.3% of the samples. Of these, cyprinids and centrarchids were collected most frequently. Proportional fish species richness estimated for the first pass increased significantly with decreasing stream width for 1 of the 10 river basins. When used to calculate probabilities of detection of individual fish species, the removal model failed 48% of the time because the number of individuals of a species was greater in the second pass than in the first pass. Single-pass backpack electrofishing data alone may make it difficult to determine whether characterized fish community structure data are real or spurious. The two-pass removal model can be used to assess the effectiveness of sampling species richness with a single electrofishing pass. However, the two-pass removal model may have limited utility to determine probabilities of detection of individual species and, thus, limit the ability to assess the effectiveness of single-pass sampling to characterize species relative abundances. Multiple-pass (at least three passes) backpack electrofishing at a large number of sites may not be cost-effective as part of a standardized sampling protocol for large-geographic-scale studies. However, multiple

Assessment of the St. Louis River AOC fish tumors and other ...

EPA Pesticide Factsheets

The Fish Tumors and Other Deformities Beneficial Use Impairment (BUI) was listed as one of nine BUIs at the time the St. Louis River AOC was designated in 1987. At the time, no formal studies had been conducted to estimate the prevalence of either fish tumors or deformities. To assess the current status of fish tumors and deformities in the AOC, adult white suckers (Catostomus commersonii) were sampled during the spawning period in May during 2011, 2013 and 2015. We measured each fish, determined its sex and age (sagittal otolith), sampled dorsal muscle tissue for carbon and nitrogen stable isotope ratios to determine river versus lake residency, and conducted a necropsy-based assessment to document grossly visible abnormalities. Pieces of any observable abnormalities were preserved for subsequent histological analyses. A total of 622 white sucker were surveyed from 2011 through 2015. Of these, 27 (4.3%) white suckers had skin neoplasms, all of which were papillomas. A total of 29 (4.7%) of the white suckers had liver neoplasms. Sex and age were significantly associated with skin neoplasia incidence, whereas only age was significantly associated with liver neoplasia incidence. Neither neoplasia type was significantly associated with habitat use, indicating that neoplasia incidence did not change with increasing feeding in the AOC relative to Lake Superior. Further, there was not a significant difference in skin or liver neoplasia incidence between migratory whit

Age-related changes in hematology and plasma chemistry values of hybrid striped bass (Morone chrysops X Morone saxatilis).

PubMed

Hrubec, Terry C.; Smith, Stephen A.; Robertson, John L.

2001-01-01

Hybrid striped bass (Morone chrysops X Morone saxatilis ) are an important aquaculture species yet there are few diagnostic tools available to assess their health. Hematology and clinical chemistry analyses are not used extensively in fish medicine due to the lack of reference intervals for various fish species, and because factors such as age can affect blood values. There is little published information regarding age-related changes in blood values of juvenile fish. It is important to evaluate juvenile fish, as this is the time they are raised in aquaculture settings. Determining age-related changes in the blood values of fishes would further develop clinical pathology as a diagnostic tool, enhancing both fish medicine and the aquaculture industry. The results of standard hematology and clinical chemistry analysis were evaluated in juvenile hybrid striped bass at 4, 6, 9, 15, and 19 months of age. Values for PCV and RBC indices were significantly lower, and plasma protein concentration was significantly higher in younger fish. Total WBC and lymphocyte counts were significantly higher in fish at 6 and 9 months of age, while neutrophil and monocyte counts were higher at 6, 9, and 15 months. Eosinophil counts were significantly higher in 9-month-old fish. The majority of hematologic values fell within previously established reference intervals, indicating that only slight modification to the intervals is necessary for evaluating hematologic results of hybrid striped bass at different ages. The following analytes deviated sufficiently from adult reference intervals to warrant separate reference values: plasma protein concentration at 4 months, WBC and lymphocyte counts at 15 and 19 months, and thrombocyte-like-cells at 9 months of age. Values for most biochemical analytes were significantly different among age groups except for creatinine and potassium concentrations. Comparisons with reference intervals were not made for biochemical analytes, because established

Assessing Fish and Motile Fauna around Offshore Windfarms Using Stereo Baited Video

PubMed Central

Griffin, Ross A.; Robinson, Gary J.; West, Ashley; Gloyne-Phillips, Ian T.; Unsworth, Richard K. F.

2016-01-01

There remains limited knowledge of how offshore windfarm developments influence fish assemblages, particularly at a local scale around the turbine structures. Considering the existing levels of anthropogenic pressures on coastal fish populations it is becoming increasingly important for developers and environmental regulators to gain a more comprehensive understanding of the factors influencing fish assemblages. Improving our ability to assess such fish populations in close proximity to structures will assist in increasing this knowledge. In the present study we provide the first trial use of Baited Remote Underwater Stereo-Video systems (stereo BRUVs) for the quantification of motile fauna in close proximity to offshore wind turbines. The study was conducted in the Irish Sea and finds the technique to be a viable means of assessing the motile fauna of such environments. The present study found a mixture of species including bottom dwellers, motile crustaceans and large predatory fish. The majority of taxa observed were found to be immature individuals with few adult individuals recorded. The most abundant species were the angular crab (Goneplax rhomboides) and the small-spotted catshark (Scyliorhinus canicula). Of note in this study was the generally low abundance and diversity of taxa recorded across all samples, we hypothesise that this reflects the generally poor state of the local fauna of the Irish Sea. The faunal assemblages sampled in close proximity to turbines were observed to alter with increasing distance from the structure, species more characteristic of hard bottom environments were in abundance at the turbines (e.g. Homarus gammarus, Cancer pagarus, Scyliorhinus spp.) and those further away more characteristic of soft bottoms (e.g. Norwegian Lobster). This study highlights the need for the environmental impacts of offshore renewables on motile fauna to be assessed using targeted and appropriate tools. Stereo BRUVs provide one of those tools, but like

The fish embryo toxicity test as an animal alternative method in hazard and risk assessment and scientific research.

PubMed

Embry, Michelle R; Belanger, Scott E; Braunbeck, Thomas A; Galay-Burgos, Malyka; Halder, Marlies; Hinton, David E; Léonard, Marc A; Lillicrap, Adam; Norberg-King, Teresa; Whale, Graham

2010-04-15

Animal alternatives research has historically focused on human safety assessments and has only recently been extended to environmental testing. This is particularly for those assays that involve the use of fish. A number of alternatives are being pursued by the scientific community including the fish embryo toxicity (FET) test, a proposed replacement alternative to the acute fish test. Discussion of the FET methodology and its application in environmental assessments on a global level was needed. With this emerging issue in mind, the ILSI Health and Environmental Sciences Institute (HESI) and the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) held an International Workshop on the Application of the Fish Embryo Test as an Animal Alternative Method in Hazard and Risk Assessment and Scientific Research in March, 2008. The workshop included approximately 40 scientists and regulators representing government, industry, academia, and non-governmental organizations from North America, Europe, and Asia. The goal was to review the state of the science regarding the investigation of fish embryonic tests, pain and distress in fish, emerging approaches utilizing fish embryos, and the use of fish embryo toxicity test data in various types of environmental assessments (e.g., hazard, risk, effluent, and classification and labeling of chemicals). Some specific key outcomes included agreement that risk assessors need fish data for decision-making, that extending the FET to include eluethereombryos was desirable, that relevant endpoints are being used, and that additional endpoints could facilitate additional uses beyond acute toxicity testing. The FET was, however, not yet considered validated sensu OECD. An important action step will be to provide guidance on how all fish tests can be used to assess chemical hazard and to harmonize the diverse terminology used in test guidelines adopted over the past decades. Use of the FET in context of effluent assessments

Asexual Reproduction Does Not Apparently Increase the Rate of Chromosomal Evolution: Karyotype Stability in Diploid and Triploid Clonal Hybrid Fish (Cobitis, Cypriniformes, Teleostei)

PubMed Central

Majtánová, Zuzana; Choleva, Lukáš; Symonová, Radka; Ráb, Petr; Kotusz, Jan; Pekárik, Ladislav; Janko, Karel

2016-01-01

Interspecific hybridization, polyploidization and transitions from sexuality to asexuality considerably affect organismal genomes. Especially the last mentioned process has been assumed to play a significant role in the initiation of chromosomal rearrangements, causing increased rates of karyotype evolution. We used cytogenetic analysis and molecular dating of cladogenetic events to compare the rate of changes of chromosome morphology and karyotype in asexually and sexually reproducing counterparts in European spined loach fish (Cobitis). We studied metaphases of three sexually reproducing species and their diploid and polyploid hybrid clones of different age of origin. The material includes artificial F1 hybrid strains, representatives of lineage originated in Holocene epoch, and also individuals of an oldest known age to date (roughly 0.37 MYA). Thereafter we applied GISH technique as a marker to differentiate parental chromosomal sets in hybrids. Although the sexual species accumulated remarkable chromosomal rearrangements after their speciation, we observed no differences in chromosome numbers and/or morphology among karyotypes of asexual hybrids. These hybrids possess chromosome sets originating from respective parental species with no cytogenetically detectable recombinations, suggesting their integrity even in a long term. The switch to asexual reproduction thus did not provoke any significant acceleration of the rate of chromosomal evolution in Cobitis. Asexual animals described in other case studies reproduce ameiotically, while Cobitis hybrids described here produce eggs likely through modified meiosis. Therefore, our findings indicate that the effect of asexuality on the rate of chromosomal change may be context-dependent rather than universal and related to particular type of asexual reproduction. PMID:26808475

Asexual Reproduction Does Not Apparently Increase the Rate of Chromosomal Evolution: Karyotype Stability in Diploid and Triploid Clonal Hybrid Fish (Cobitis, Cypriniformes, Teleostei).

PubMed

Majtánová, Zuzana; Choleva, Lukáš; Symonová, Radka; Ráb, Petr; Kotusz, Jan; Pekárik, Ladislav; Janko, Karel

2016-01-01

Interspecific hybridization, polyploidization and transitions from sexuality to asexuality considerably affect organismal genomes. Especially the last mentioned process has been assumed to play a significant role in the initiation of chromosomal rearrangements, causing increased rates of karyotype evolution. We used cytogenetic analysis and molecular dating of cladogenetic events to compare the rate of changes of chromosome morphology and karyotype in asexually and sexually reproducing counterparts in European spined loach fish (Cobitis). We studied metaphases of three sexually reproducing species and their diploid and polyploid hybrid clones of different age of origin. The material includes artificial F1 hybrid strains, representatives of lineage originated in Holocene epoch, and also individuals of an oldest known age to date (roughly 0.37 MYA). Thereafter we applied GISH technique as a marker to differentiate parental chromosomal sets in hybrids. Although the sexual species accumulated remarkable chromosomal rearrangements after their speciation, we observed no differences in chromosome numbers and/or morphology among karyotypes of asexual hybrids. These hybrids possess chromosome sets originating from respective parental species with no cytogenetically detectable recombinations, suggesting their integrity even in a long term. The switch to asexual reproduction thus did not provoke any significant acceleration of the rate of chromosomal evolution in Cobitis. Asexual animals described in other case studies reproduce ameiotically, while Cobitis hybrids described here produce eggs likely through modified meiosis. Therefore, our findings indicate that the effect of asexuality on the rate of chromosomal change may be context-dependent rather than universal and related to particular type of asexual reproduction.

Apparent digestibility of Asian carp and common carp-derived fish meals in feeds for hybrid striped bass Morone saxatilis female x M. chrysops male and rainbow trout Oncorhynchus mykiss

USDA-ARS?s Scientific Manuscript database

Apparent digestibility coefficients (ADCs) of nutrients (crude protein, amino acids, crude lipid, fatty acids, and minerals) were determined for fish meals derived from menhaden, Asian carp (combination of silver and bighead carps), and common carp in feeds for hybrid striped bass and rainbow trout....

Human epidermal growth factor receptor 2 assessment in a case-control study: comparison of fluorescence in situ hybridization and quantitative reverse transcription polymerase chain reaction performed by central laboratories.

PubMed

Baehner, Frederick L; Achacoso, Ninah; Maddala, Tara; Shak, Steve; Quesenberry, Charles P; Goldstein, Lynn C; Gown, Allen M; Habel, Laurel A

2010-10-01

The optimal method to assess human epidermal growth factor receptor 2 (HER2) status remains highly controversial. Before reporting patient HER2 results, American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines mandate that laboratories demonstrate ≥ 95% concordance to another approved laboratory or methodology. Here, we compare central laboratory HER2 assessed by fluorescence in situ hybridization (FISH) and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using Oncotype DX in lymph node-negative, chemotherapy-untreated patients from a large Kaiser Permanente case-control study. Breast cancer specimens from the Kaiser-Genomic Health study were examined. Central FISH assessment of HER2 amplification and polysomy 17 was conducted by PhenoPath Laboratories (ratios > 2.2, 1.8 to 2.2, and < 1.8 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). HER2 expression by RT-PCR was conducted using Oncotype DX by Genomic Health (normalized expression units ≥ 11.5, 10.7 to < 11.5, and < 10.7 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). Concordance analyses followed ASCO/CAP guidelines. HER2 concordance by central FISH and central RT-PCR was 97% (95% CI, 96% to 99%). Twelve percent (67 of 568 patients) and 11% (60 of 568 patients) of patients were HER2 positive by RT-PCR and FISH, respectively. HER2-positive patients had increased odds of dying from breast cancer compared with HER2-negative patients. Polysomy 17 was demonstrated in 12.5% of all patients and 33% of FISH-positive patients. Nineteen of 20 FISH-positive patients with polysomy 17 were also RT-PCR HER2 positive. Although not statistically significantly different, HER2-positive/polysomy 17 patients tended to have the worst prognosis, followed by HER2-positive/eusomic, HER2-negative/polysomy 17, and HER2-negative/eusomic patients. There is a high degree of concordance between central FISH and quantitative RT

Using field data to assess the tolerance of freshwater fish to elevated ionic concentrations

EPA Science Inventory

We used field data of fish occurrences and specific conductivity to assess the tolerance of freshwater fish to elevated ions. The concentration at which a species was expected to no longer be observed [the extirpation concentration (XC95)] was identified from the 95th percentile ...

Probabilistic risk assessment of exposure to leucomalachite green residues from fish products.

PubMed

Chu, Yung-Lin; Chimeddulam, Dalaijamts; Sheen, Lee-Yan; Wu, Kuen-Yuh

2013-12-01

To assess the potential risk of human exposure to carcinogenic leucomalachite green (LMG) due to fish consumption, the probabilistic risk assessment was conducted for adolescent, adult and senior adult consumers in Taiwan. The residues of LMG with the mean concentration of 13.378±20.56 μg kg(-1) (BFDA, 2009) in fish was converted into dose, considering fish intake reported for three consumer groups by NAHSIT (1993-1996) and body weight of an average individual of the group. The lifetime average and high 95th percentile dietary intakes of LMG from fish consumption for Taiwanese consumers were estimated at up to 0.0135 and 0.0451 μg kg-bw(-1) day(-1), respectively. Human equivalent dose (HED) of 2.875 mg kg-bw(-1) day(-1) obtained from a lower-bound benchmark dose (BMDL10) in mice by interspecies extrapolation was linearly extrapolated to oral cancer slope factor (CSF) of 0.035 (mgkg-bw(-1)day(-1))(-1) for humans. Although, the assumptions and methods are different, the results of lifetime cancer risk varying from 3×10(-7) to 1.6×10(-6) were comparable to those of margin of exposures (MOEs) varying from 410,000 to 4,800,000. In conclusions, Taiwanese fish consumers with the 95th percentile LADD of LMG have greater risk of liver cancer and need to an action of risk management in Taiwan. Copyright © 2013 Elsevier Ltd. All rights reserved.

Assessing the impact of fishing in shallow rocky reefs: a multivariate approach to ecosystem management.

PubMed

Sangil, Carlos; Martín-García, Laura; Clemente, Sabrina

2013-11-15

In this paper we develop a tool to assess the impact of fishing on ecosystem functioning in shallow rocky reefs. The relationships between biological parameters (fishes, sea urchins, seaweeds), and fishing activities (fish traps, boats, land-based fishing, spearfishing) were tested in La Palma island (Canary Islands). Data from fishing activities and biological parameters were analyzed using principal component analyses. We produced two models using the first component of these analyses. This component was interpreted as a new variable that described the fishing pressure and the conservation status at each studied site. Subsequently the scores on the first axis were mapped using universal kriging methods and the models obtained were extrapolated across the whole island to display the expected fishing pressure and conservation status more widely. The fishing pressure and conservation status models were spatially related; zones where fishing pressure was high coincided with zones in the unhealthiest ecological state. Copyright © 2013 Elsevier Ltd. All rights reserved.

Assessing ecological risks to the fish community from residual coal fly ash in Watts Bar Reservoir, Tennessee

DOE PAGES

Rigg, David K.; Wacksman, Mitch N.; Iannuzzi, Jacqueline; ...

2014-12-18

For this research, extensive site-specific biological and environmental data were collected to support an evaluation of risks to the fish community in Watts Bar Reservoir from residual ash from the December 2008 Tennessee Valley Authority (TVA) Kingston ash release. This paper describes the approach used and results of the risk assessment for the fish community, which consists of multiple measurement endpoints (measures of exposure and effects) for fish. The lines of evidence included 1) comparing postspill annual fish community assessments with nearby prespill data and data from other TVA reservoirs, 2) evaluating possible effects of exposures of fish eggs andmore » larval fish to ash in controlled laboratory toxicity tests, 3) evaluating reproductive competence of field-exposed fish, 4) assessing individual fish health through physical examination, histopathology, and blood chemistry, 5) comparing fish tissue concentrations with literature-based critical body residues, and 6) comparing concentrations of ash-related contaminants in surface waters with US Environmental Protection Agency's (USEPA) Ambient Water Quality Standards for Fish and Aquatic Life. These measurement endpoints were treated as independent lines of evidence that were integrated into an overall weight-of-evidence estimate of risk to the fish community. Collectively, the data and analysis presented here indicate that ash and ash-related constituents pose negligible risks to the fish communities in Watts Bar Reservoir. This conclusion contradicts the predictions by some researchers immediately following the ash release of devastating effects on the aquatic ecology of Watts Bar Reservoir. The information presented in this article reaffirms the wisdom of carefully evaluating the evidence before predicting probable ecological effects of a major event such as the TVA Kingston ash release. Lastly, this study demonstrates that a thorough and detailed investigation using multiple measurement endpoints

Assessing ecological risks to the fish community from residual coal fly ash in Watts Bar Reservoir, Tennessee

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rigg, David K.; Wacksman, Mitch N.; Iannuzzi, Jacqueline

For this research, extensive site-specific biological and environmental data were collected to support an evaluation of risks to the fish community in Watts Bar Reservoir from residual ash from the December 2008 Tennessee Valley Authority (TVA) Kingston ash release. This paper describes the approach used and results of the risk assessment for the fish community, which consists of multiple measurement endpoints (measures of exposure and effects) for fish. The lines of evidence included 1) comparing postspill annual fish community assessments with nearby prespill data and data from other TVA reservoirs, 2) evaluating possible effects of exposures of fish eggs andmore » larval fish to ash in controlled laboratory toxicity tests, 3) evaluating reproductive competence of field-exposed fish, 4) assessing individual fish health through physical examination, histopathology, and blood chemistry, 5) comparing fish tissue concentrations with literature-based critical body residues, and 6) comparing concentrations of ash-related contaminants in surface waters with US Environmental Protection Agency's (USEPA) Ambient Water Quality Standards for Fish and Aquatic Life. These measurement endpoints were treated as independent lines of evidence that were integrated into an overall weight-of-evidence estimate of risk to the fish community. Collectively, the data and analysis presented here indicate that ash and ash-related constituents pose negligible risks to the fish communities in Watts Bar Reservoir. This conclusion contradicts the predictions by some researchers immediately following the ash release of devastating effects on the aquatic ecology of Watts Bar Reservoir. The information presented in this article reaffirms the wisdom of carefully evaluating the evidence before predicting probable ecological effects of a major event such as the TVA Kingston ash release. Lastly, this study demonstrates that a thorough and detailed investigation using multiple measurement endpoints

The 2014 FDA assessment of commercial fish: practical considerations for improved dietary guidance.

PubMed

McGuire, Jennifer; Kaplan, Jason; Lapolla, John; Kleiner, Rima

2016-07-13

The U.S. Food and Drug Administration (FDA) recently released its report: A Quantitative Assessment of the Net Effects on Fetal Neurodevelopment from Eating Commercial Fish (As Measured by IQ and also by Early Age Verbal Development in Children). By evaluating the benefits and potential concerns of eating fish during pregnancy and breastfeeding, the analysis suggests that pregnant women consuming two seafood meals (8-12 oz) per week could provide their child with an additional 3.3 IQ points by age 9. Recent insights from behavioral economics research indicate that other factors, such as concerns about price and methylmercury (MeHg) exposure, appear to reduce fish consumption in many individuals.To assess the net effects of eating commercial fish during pregnancy, we compared the consumption of select fish species necessary to achieve IQ benefits with the amount necessary to have adverse developmental effects due to MeHg exposure. For the species or market types evaluated, the number of servings necessary to reach MeHg exposure to observe an adverse effect was at least twice that the amount estimated to achieve peak developmental benefit. We then reported average costs of fresh and canned or pouched fish, and calculated the cost per week for pregnant women to achieve maximum IQ benefits for their gestating child. Canned light tuna was the least expensive option at $1.83 per week to achieve maximum IQ benefit.Due to their relatively low cost, canned and pouched fish products eaten with enough regularity are likely to provide peak cognitive benefits. Because of its popularity, canned and pouched tuna could provide some of the largest cognitive benefits from fish consumption in the U.S. Future FDA consumer advice and related educational initiatives could benefit from a broader perspective that highlights the importance of affordable and accessible fish choices. These observations underscore the importance of clear public health messaging that address both health

Contaminant exposure in Montrealers of Asian origin fishing the St. Lawrence River: Exploratory assessment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kosatsky, T.; Przybysz, R.; Shatenstein, B.

1999-02-01

Fishing and fish consumption are widely practiced among members of certain ethnocultural groups. Information assessment led them to ascribe high levels of consumption of locally caught sportfish to Montrealers of Asian origin and to hypothesize that their choices of species and fish organs differ from those of the majority group. An exploratory assessment of contaminant bioindicators reflective of St. Lawrence river fish consumption was conducted in late 1995 among nine Vietnamese and nine Bangladeshi Montreal sportfishers identified by community contacts. Vietnamese participants, six men and three women, were 27--70 years of age and had immigrated to Canada 3--20 years earlier.more » In contrast, the nine Bangladeshi males aged 28--41 years had been in Canada for 2--13 years. Bioindicator concentrations among Bangladeshi and Vietnamese participants are compared to those found for majority-community Montreal-area high-level consumers recruited on the St. Lawrence River during winter 1995.« less

Assessing ecological water quality with macroinvertebrates and fish: a case study from a small Mediterranean river.

PubMed

Cheimonopoulou, Maria Th; Bobori, Dimitra C; Theocharopoulos, Ioannis; Lazaridou, Maria

2011-02-01

Biological elements, such as benthic macroinvertebrates and fish, have been used in assessing the ecological quality of rivers according to the requirements of the Water Framework Directive. However, the concurrent use of multiple organism groups provides a broader perspective for such evaluations, since each biological element may respond differently to certain environmental variables. In the present study, we assessed the ecological quality of a Greek river (RM4 type), during autumn 2003 and spring 2004 at 10 sites, with benthic macroinvertebrates and fish. Hydromorphological and physicochemical parameters, habitat structure, and riparian vegetation were also considered. Pollution sensitive macroinvertebrate taxa were more abundant at headwaters, which had good/excellent water quality according to the Hellenic Evaluation System (HES). The main river reaches possessed moderate water quality, while downstream sites were mainly characterised as having bad or poor water quality, dominated by pollution-tolerant macroinvertebrate taxa. Macroinvertebrates related strongly to local stressors as chemical degradation (ordination analysis CCA) and riparian quality impairment (bivariate analysis) while fish did not. Fish were absent from the severely impacted lower river reaches. Furthermore, external pathological signs were observed in fish caught at certain sites. A combined use of both macroinvertebrates and fish in biomonitoring programs is proposed for providing a safer assessment of local and regional habitat impairment.

Assessing the techno-economics of modular hybrid solar thermal systems

NASA Astrophysics Data System (ADS)

Lim, Jin Han; Chinnici, Alfonso; Dally, Bassam; Nathan, Graham

2017-06-01

A techno-economic assessment was performed on modular hybrid solar thermal (in particular, solar power tower) systems with combustion from natural gas as backup to provide a continuous supply of electricity. Two different configurations were compared, i.e. a Hybrid Solar Receiver Combustor (HSRC), in which the functions of a solar cavity receiver and a combustor are integrated into a single device, and a Solar Gas Hybrid (SGH), which is a reference hybrid solar thermal system with a standalone solar-only cavity receiver and a backup boiler. The techno-economic benefits were assessed by varying the size of the modular components, i.e. the heliostat field and the solar receivers. It was found that for modularization to be cost effective requires more than the increased learning from higher production of a larger number of smaller units, such as access to alternative, lower-cost manufacturing methods and/or the use of a low melting point Heat Transfer Fluid (HTF) such as sodium to reduce parasitic losses. In particular, for a plant with 30 units of 1MWth modules, the Levelized Cost of Electricity is competitive compared with a single unit of 30MWth after ˜100 plants are installed for both the HSRC and SGH if the systems employ the use of sodium as the heat transfer fluid.

Assessment of injury to fish and wildlife resources in the Grand Calumet River and Indiana Harbor Area of Concern, USA

USGS Publications Warehouse

MacDonald, D.D.; Ingersoll, C.G.; Smorong, D.E.; Lindskoog, R.A.; Sparks, D.W.; Smith, J.R.; Simon, T.P.; Hanacek, M.A.

2002-01-01

This article is the second in a series of three that describes the results of a Natural Resource Damage Assessment (NRDA) conducted in the Grand Calumet River and Indiana Harbor Area of Concern (IHAOC). The assessment area is located in northwest Indiana and was divided into nine reaches to facilitate the assessment. This component of the NRDA was undertaken to determine if fish and wildlife resources have been injured due to exposure to contaminants that are associated with discharges of oil or releases of other hazardous substances. To support this assessment, information was compiled on the chemical composition of sediment and tissues; on the toxicity of whole sediments, pore water, and elutriates to fish; on the status of fish communities; and on fish health. The data on each of these indicators were compared to regionally relevant benchmarks to assess the presence and extent of injury to fish and wildlife resources. The results of this assessment indicate that injury to fish and wildlife resources has occurred throughout the assessment area, with up to five distinct lines of evidence demonstrating injury within the various reaches. Based on the frequency of exceedance of the benchmarks for assessing sediment and tissue chemistry data, total polychlorinated biphenyls is the primary bioaccumulative contaminant of concern in the assessment area. It is important to note, however, that this assessment was restricted by the availability of published bioaccumulation-based sediment quality guidelines, tissue residue guidelines, and other benchmarks of sediment quality conditions. The availability of chemistry data for tissues also restricted this assessment in certain reaches of the assessment area. Furthermore, insufficient information was located to facilitate identification of the substances that are causing or substantially contributing to effects on fish (i.e., sediment toxicity, impaired fish health, or impaired fish community structure). Therefore, substances

Combination of Fluorescent in situ Hybridization (FISH) and Immunofluorescence Imaging for Detection of Cytokine Expression in Microglia/Macrophage Cells

PubMed Central

Fe Lanfranco, Maria; Loane, David J.; Mocchetti, Italo; Burns, Mark P.; Villapol, Sonia

2017-01-01

Microglia and macrophage cells are the primary producers of cytokines in response to neuroinflammatory processes. But these cytokines are also produced by other glial cells, endothelial cells, and neurons. It is essential to identify the cells that produce these cytokines to target their different levels of activation. We used dual RNAscope® fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) techniques to visualize the mRNA expression pattern of pro- and anti-inflammatory cytokines in microglia/macrophages cells. Using these methods, we can associate one mRNA to specific cell types when combining with different cellular markers by immunofluorescence. Results from RNAscope® probes IL-1β, TNFα, TGFβ, IL-10 or Arg1, showed colocalization with antibodies for microglia/macrophage cells. These target probes showed adequate sensitivity and specificity to detect mRNA expression. New FISH detection techniques combined with immunohistochemical techniques will help to jointly determine the protein and mRNA localization, as well as provide reliable quantification of the mRNA expression levels. PMID:29238736

Shoal Bass hybridization below Morgan Falls Dam.

USGS Publications Warehouse

Taylor, Andrew T.; Tringali, Michael D.; O'Rourke, Patrick M.; Long, James M.

2018-01-01

The shoal bass (Micropterus cataractae) is a sportfish endemic to the Apalachicola-Chattahoochee-Flint Basin of the southeastern United States. Introgression with several non-native congeners poses a pertinent threat to shoal bass conservation, particularly in the altered habitats of the Chattahoochee River. Our primary objective was to characterize hybridization in shoal bass populations near Atlanta, Georgia, including a population inhabiting Big Creek and another in the main stem Chattahoochee River below Morgan Falls Dam (MFD). A secondary objective was to examine the accuracy of phenotypic identifications below MFD based on a simplified suite of characters examined in the field. Fish were genotyped with 16 microsatellite DNA markers, and results demonstrated that at least four black bass species were involved in introgressive hybridization. Of 62 fish genotyped from Big Creek, 27% were pure shoal bass and 65% represented either F1 hybrids of shoal bass x smallmouth bass (M. dolomieu) or unidirectional backcrosses towards shoal bass. Of 29 fish genotyped below MFD and downstream at Cochran Shoals, 45% were pure shoal bass. Six hybrid shoal bass included both F1 hybrids and backcrosses with non-natives including Alabama bass (M. henshalli), spotted bass (M. punctulatus), and smallmouth bass. Shoal bass alleles comprised only 21% of the overall genomic composition in Big Creek and 31% below MFD (when combined with Cochran Shoals). Phenotypic identification below MFD resulted in an overall correct classification rate of 86% when discerning pure shoal bass from all other non-natives and hybrids. Results suggest that although these two shoal bass populations feature some of the highest introgression rates documented, only a fleeting opportunity may exist to conserve pure shoal bass in both populations. Continued supplemental stocking of pure shoal bass below MFD appears warranted to thwart increased admixture among multiple black bass taxa, and a similar stocking

MYC and MYCN amplification can be reliably assessed by aCGH in medulloblastoma.

PubMed

Bourdeaut, Franck; Grison, Camille; Maurage, Claude-Alain; Laquerriere, Annie; Vasiljevic, Alexandre; Delisle, Marie-Bernadette; Michalak, Sophie; Figarella-Branger, Dominique; Doz, François; Richer, Wilfrid; Pierron, Gaelle; Miquel, Catherine; Delattre, Olivier; Couturier, Jérôme

2013-04-01

As prognostic factors, MYC and MYCN amplifications are routinely assessed in medulloblastomas. Fluorescence in situ hybridization (FISH) is currently considered as the technique of reference. Recently, array comparative genomic hybridization (aCGH) has been developed as an alternative technique to evaluate genomic abnormalities in other tumor types; however, this technique has not been widely adopted as a replacement for FISH in medulloblastoma. In this study, 34 tumors were screened by both FISH and aCGH. In all cases showing amplification by FISH, aCGH also unambiguously revealed the abnormality. The aCGH technique was also performed on tumors showing no amplification by FISH, and the absence of amplification was confirmed in all cases. Interestingly, one tumor showed a subclonal MYC amplification by FISH. This subclonal amplification was observed in approximately 20% of tumor cells and was clearly evident on aCGH. In conclusion, our analysis confirms that aCGH is as safe as FISH for the detection of MYC/MYCN gene amplification. Given its cost efficiency in comparison to two FISH tests and the global genomic information additionally provided by an aCGH experiment, this reproducible technique can be safely retained as an alternative to FISH for routine investigation of medulloblastoma. Copyright © 2013 Elsevier Inc. All rights reserved.

Customized Oligonucleotide Array-Based Comparative Genomic Hybridization as a Clinical Assay for Genomic Profiling of Chronic Lymphocytic Leukemia

PubMed Central

Sargent, Rachel; Jones, Dan; Abruzzo, Lynne V.; Yao, Hui; Bonderover, Jaime; Cisneros, Marissa; Wierda, William G.; Keating, Michael J.; Luthra, Rajyalakshmi

2009-01-01

Chromosome gains and losses used for risk stratification in chronic lymphocytic leukemia (CLL) are commonly assessed by multiprobe fluorescence in situ hybridization (FISH) studies. We designed and validated a customized array-comparative genomic hybridization (aCGH) platform as a clinical assay for CLL genomic profiling. A 60-mer, 44,000-probe oligonucleotide array with a 50-kb average spatial resolution was augmented with high-density probe tiling at loci that are frequently aberrant in CLL. Aberrations identified by aCGH were compared with those identified by a FISH panel, including locus-specific probes to ATM (11q22.3), the centromeric region of chromosome 12 (12p11.1–q11), D13S319 (13q14.3), LAMP1 (13q34), and TP53 (17p13.1). In 100 CLL samples, aCGH/FISH concordance was seen for 89% of FISH-called aberrations at the ATM (n = 18), D13S319 (n = 42), LAMP (n = 12), and TP53 (n = 22) loci and for chromosome 12 (n = 14). Eighty-four percentage of FISH/aCGH discordant calls were in samples either at or below the limit of aCGH sensitivity (10% to 25% FISH aberration-containing cells). Therefore, aCGH profiling is a feasible routine clinical test with comparable results to multiprobe FISH studies; however, it may be less sensitive than FISH in cases with low-level aberrations. Further, a customized array design can provide comprehensive genomic profiling with additional accuracy in both identifying and defining the extent of small aberrations at target loci. PMID:19074592

ZyFISH: A Simple, Rapid and Reliable Zygosity Assay for Transgenic Mice

PubMed Central

McHugh, Donal; O’Connor, Tracy; Bremer, Juliane; Aguzzi, Adriano

2012-01-01

Microinjection of DNA constructs into fertilized mouse oocytes typically results in random transgene integration at a single genomic locus. The resulting transgenic founders can be used to establish hemizygous transgenic mouse lines. However, practical and experimental reasons often require that such lines be bred to homozygosity. Transgene zygosity can be determined by progeny testing assays which are expensive and time-consuming, by quantitative Southern blotting which is labor-intensive, or by quantitative PCR (qPCR) which requires transgene-specific design. Here, we describe a zygosity assessment procedure based on fluorescent in situ hybridization (zyFISH). The zyFISH protocol entails the detection of transgenic loci by FISH and the concomitant assignment of homozygosity using a concise and unbiased scoring system. The method requires small volumes of blood, is scalable to at least 40 determinations per assay, and produces results entirely consistent with the progeny testing assay. This combination of reliability, simplicity and cost-effectiveness makes zyFISH a method of choice for transgenic mouse zygosity determinations. PMID:22666404

Climate change vulnerability of native and alien freshwater fishes of California: a systematic assessment approach.

PubMed

Moyle, Peter B; Kiernan, Joseph D; Crain, Patrick K; Quiñones, Rebecca M

2013-01-01

Freshwater fishes are highly vulnerable to human-caused climate change. Because quantitative data on status and trends are unavailable for most fish species, a systematic assessment approach that incorporates expert knowledge was developed to determine status and future vulnerability to climate change of freshwater fishes in California, USA. The method uses expert knowledge, supported by literature reviews of status and biology of the fishes, to score ten metrics for both (1) current status of each species (baseline vulnerability to extinction) and (2) likely future impacts of climate change (vulnerability to extinction). Baseline and climate change vulnerability scores were derived for 121 native and 43 alien fish species. The two scores were highly correlated and were concordant among different scorers. Native species had both greater baseline and greater climate change vulnerability than did alien species. Fifty percent of California's native fish fauna was assessed as having critical or high baseline vulnerability to extinction whereas all alien species were classified as being less or least vulnerable. For vulnerability to climate change, 82% of native species were classified as highly vulnerable, compared with only 19% for aliens. Predicted climate change effects on freshwater environments will dramatically change the fish fauna of California. Most native fishes will suffer population declines and become more restricted in their distributions; some will likely be driven to extinction. Fishes requiring cold water (<22°C) are particularly likely to go extinct. In contrast, most alien fishes will thrive, with some species increasing in abundance and range. However, a few alien species will likewise be negatively affected through loss of aquatic habitats during severe droughts and physiologically stressful conditions present in most waterways during summer. Our method has high utility for predicting vulnerability to climate change of diverse fish species. It

Climate Change Vulnerability of Native and Alien Freshwater Fishes of California: A Systematic Assessment Approach

PubMed Central

Moyle, Peter B.; Kiernan, Joseph D.; Crain, Patrick K.; Quiñones, Rebecca M.

2013-01-01

Freshwater fishes are highly vulnerable to human-caused climate change. Because quantitative data on status and trends are unavailable for most fish species, a systematic assessment approach that incorporates expert knowledge was developed to determine status and future vulnerability to climate change of freshwater fishes in California, USA. The method uses expert knowledge, supported by literature reviews of status and biology of the fishes, to score ten metrics for both (1) current status of each species (baseline vulnerability to extinction) and (2) likely future impacts of climate change (vulnerability to extinction). Baseline and climate change vulnerability scores were derived for 121 native and 43 alien fish species. The two scores were highly correlated and were concordant among different scorers. Native species had both greater baseline and greater climate change vulnerability than did alien species. Fifty percent of California’s native fish fauna was assessed as having critical or high baseline vulnerability to extinction whereas all alien species were classified as being less or least vulnerable. For vulnerability to climate change, 82% of native species were classified as highly vulnerable, compared with only 19% for aliens. Predicted climate change effects on freshwater environments will dramatically change the fish fauna of California. Most native fishes will suffer population declines and become more restricted in their distributions; some will likely be driven to extinction. Fishes requiring cold water (<22°C) are particularly likely to go extinct. In contrast, most alien fishes will thrive, with some species increasing in abundance and range. However, a few alien species will likewise be negatively affected through loss of aquatic habitats during severe droughts and physiologically stressful conditions present in most waterways during summer. Our method has high utility for predicting vulnerability to climate change of diverse fish species. It

Bioelectrical impedance analysis: A new tool for assessing fish condition

USGS Publications Warehouse

Hartman, Kyle J.; Margraf, F. Joseph; Hafs, Andrew W.; Cox, M. Keith

2015-01-01

Bioelectrical impedance analysis (BIA) is commonly used in human health and nutrition fields but has only recently been considered as a potential tool for assessing fish condition. Once BIA is calibrated, it estimates fat/moisture levels and energy content without the need to kill fish. Despite the promise held by BIA, published studies have been divided on whether BIA can provide accurate estimates of body composition in fish. In cases where BIA was not successful, the models lacked the range of fat levels or sample sizes we determined were needed for model success (range of dry fat levels of 29%, n = 60, yielding an R2 of 0.8). Reduced range of fat levels requires an increased sample size to achieve that benchmark; therefore, standardization of methods is needed. Here we discuss standardized methods based on a decade of research, identify sources of error, discuss where BIA is headed, and suggest areas for future research.

Determination of 24 personal care products in fish bile using hybrid solvent precipitation and dispersive solid phase extraction cleanup with ultrahigh performance liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry.

PubMed

Yao, Li; Lv, Yin-Zhi; Zhang, Li-Juan; Liu, Wang-Rong; Zhao, Jian-Liang; Liu, You-Sheng; Zhang, Qian-Qian; Ying, Guang-Guo

2018-05-25

Personal care products (PCPs) are ubiquitous in aquatic environments owing to the continuous discharge of domestic wastewater from highly urbanized regions. These PCPs can be adsorbed by fish and thereafter usually enter the bile of the fish through biliary excretion. In this study, a sensitive method based on a combination of hybrid solvent precipitation and dispersive solid phase extraction (d-SPE) purification was developed to simultaneously extract and detect 24 PCPs, namely, 16 biocides, 4 synthetic musks, and 4 benzotriazoles, from fish bile. Hybrid precipitation on solid phase extraction (SPE) tubes was applied to remove phospholipids and proteins, and a d-SPE procedure was used for further purification. The extraction solvents for the hybrid precipitation/SPE tubes and d-SPE materials were optimized. The method performance for bile samples both with and without enzyme hydrolysis using β-glucuronidase/aryl-sulfatase were validated. The 24 PCPs in fish bile were spiked with standard concentrations of 10 ng/mL, 20 ng/mL, 100 ng/mL, and 200 ng/mL to evaluate recoveries, which ranged from 70 to 120% for 16, 16, 22, and 21 analytes with hydrolysis, respectively, and 70-120% for 14, 15, 23, and 23 analytes without hydrolysis, respectively. The quantification limits for target PCPs were in the range 0.26-7.38 ng/mL [excluding musk xylene (MX) and musk ketone (MK)] and 0.20-9.48 ng/mL (excluding MX and MK) for bile samples with and without enzyme hydrolysis, respectively. After enzyme hydrolysis, 12 PCPs were detected in bile from fish collected from the Yangtze River, with a maximum detected concentration of 460 ng/mL, for triclosan (TCS). The hydrolysis reaction indicated that high percentages of glucuronide and sulfate metabolites for some PCPs, i.e. four parabens and TCS, existed in the bile. Copyright © 2018 Elsevier B.V. All rights reserved.

High-resolution chromosome mapping of BACs using multi-colour FISH and pooled-BAC FISH as a backbone for sequencing tomato chromosome 6.

PubMed

Szinay, Dóra; Chang, Song-Bin; Khrustaleva, Ludmila; Peters, Sander; Schijlen, Elio; Bai, Yuling; Stiekema, Willem J; van Ham, Roeland C H J; de Jong, Hans; Klein Lankhorst, René M

2008-11-01

Within the framework of the International Solanaceae Genome Project, the genome of tomato (Solanum lycopersicum) is currently being sequenced. We follow a 'BAC-by-BAC' approach that aims to deliver high-quality sequences of the euchromatin part of the tomato genome. BACs are selected from various libraries of the tomato genome on the basis of markers from the F2.2000 linkage map. Prior to sequencing, we validated the precise physical location of the selected BACs on the chromosomes by five-colour high-resolution fluorescent in situ hybridization (FISH) mapping. This paper describes the strategies and results of cytogenetic mapping for chromosome 6 using 75 seed BACs for FISH on pachytene complements. The cytogenetic map obtained showed discrepancies between the actual chromosomal positions of these BACs and their markers on the linkage group. These discrepancies were most notable in the pericentromere heterochromatin, thus confirming previously described suppression of cross-over recombination in that region. In a so called pooled-BAC FISH, we hybridized all seed BACs simultaneously and found a few large gaps in the euchromatin parts of the long arm that are still devoid of seed BACs and are too large for coverage by expanding BAC contigs. Combining FISH with pooled BACs and newly recruited seed BACs will thus aid in efficient targeting of novel seed BACs into these areas. Finally, we established the occurrence of repetitive DNA in heterochromatin/euchromatin borders by combining BAC FISH with hybridization of a labelled repetitive DNA fraction (Cot-100). This strategy provides an excellent means to establish the borders between euchromatin and heterochromatin in this chromosome.

Rapid prenatal diagnosis of chromosomal aneuploidies by fluorescence in situ hybridization: clinical experience with 4,500 specimens.

PubMed Central

Ward, B E; Gersen, S L; Carelli, M P; McGuire, N M; Dackowski, W R; Weinstein, M; Sandlin, C; Warren, R; Klinger, K W

1993-01-01

Detection of chromosome aneuploidies in uncultured amniocytes is possible using fluorescence in situ hybridization (FISH). We herein describe the results of the first clinical program which utilized FISH for the rapid detection of chromosome aneuploidies in uncultured amniocytes. FISH was performed on physician request, as an adjunct to cytogenetics in 4,500 patients. Region-specific DNA probes to chromosomes 13, 18, 21, X, and Y were used to determine ploidy by analysis of signal number in hybridized nuclei. A sample was considered to be euploid when all autosomal probes generated two hybridization signals and when a normal sex chromosome pattern was observed in greater than or equal to 80% of hybridized nuclei. A sample was considered to be aneuploid when greater than or equal to 70% of hybridized nuclei displayed the same abnormal hybridization pattern for a specific probe. Of the attempted analyses, 90.2% met these criteria and were reported as informative to referring physicians within 2 d of receipt. Based on these reporting parameters, the overall detection rate for aneuploidies was 73.3% (107/146), with an accuracy of informative results for aneuploidies of 93.9% (107/114). Compared to cytogenetics, the accuracy of all informative FISH results, euploid and aneuploid, was 99.8%, and the specificity was 99.9%. In those pregnancies where fetal abnormalities had been observed by ultrasound, referring physicians requested FISH plus cytogenetics at a significantly higher rate than they requested cytogenetics alone. The current prenatal FISH protocol is not designed to detect all chromosome abnormalities and should only be utilized as an adjunctive test to cytogenetics. This experience demonstrates that FISH can provide a rapid and accurate clinical method for prenatal identification of chromosome aneuploidies. PMID:8488836

Some potentials and limits of the leucocrit test as a fish health assessment method

USGS Publications Warehouse

Wedemeyer, G.A.; Gould, R.W.; Yasutake, W.T.

1983-01-01

The sensitivity of the leucocrit as a stress tolerance and fish health assessment method was evaluated by subjecting juvenile coho salmon, Oncorhynchus kisutch, or steelhead trout, Salmo gairdneri, to standardized crowding, handling, temperature and disease challenges. The leucocrit was a sensitive indicator of the physiological stress resulting from crowding at population densities of 0·2–0·4 kg l−1, and to the stress of handling and to temperature changes. It was relatively insensitive to physiological sampling procedures which supports its continued development as a stress assessment method.In the case of fish diseases, subclinical or active Renibacterium salmoninarum and Yersinia ruckeriinfections had essentially no effect on leucocrit values. In contrast, active Aeromonas salmonicidainfections significantly depressed the leucocrit. However, no change was seen during the subclinical (incubation) phase prior to the development of an epizootic. Thus, the potential of the leucocrit as a fish health assessment method appears limited.

A multi-scale GIS and hydrodynamic modelling approach to fish passage assessment: Clarence and Shoalhaven Rivers, NSW Australia

NASA Astrophysics Data System (ADS)

Bonetti, Rita M.; Reinfelds, Ivars V.; Butler, Gavin L.; Walsh, Chris T.; Broderick, Tony J.; Chisholm, Laurie A.

2016-05-01

Natural barriers such as waterfalls, cascades, rapids and riffles limit the dispersal and in-stream range of migratory fish, yet little is known of the interplay between these gradient dependent landforms, their hydraulic characteristics and flow rates that facilitate fish passage. The resurgence of dam construction in numerous river basins world-wide provides impetus to the development of robust techniques for assessment of the effects of downstream flow regime changes on natural fish passage barriers and associated consequences as to the length of rivers available to migratory species. This paper outlines a multi-scale technique for quantifying the relative magnitude of natural fish passage barriers in river systems and flow rates that facilitate passage by fish. First, a GIS-based approach is used to quantify channel gradients for the length of river or reach under investigation from a high resolution DEM, setting the magnitude of identified passage barriers in a longer context (tens to hundreds of km). Second, LiDAR, topographic and bathymetric survey-based hydrodynamic modelling is used to assess flow rates that can be regarded as facilitating passage across specific barriers identified by the river to reach scale gradient analysis. Examples of multi-scale approaches to fish passage assessment for flood-flow and low-flow passage issues are provided from the Clarence and Shoalhaven Rivers, NSW, Australia. In these river systems, passive acoustic telemetry data on actual movements and migrations by Australian bass (Macquaria novemaculeata) provide a means of validating modelled assessments of flow rates associated with successful fish passage across natural barriers. Analysis of actual fish movements across passage barriers in these river systems indicates that two dimensional hydraulic modelling can usefully quantify flow rates associated with the facilitation of fish passage across natural barriers by a majority of individual fishes for use in management

Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications.

PubMed

Cui, Chenghua; Shu, Wei; Li, Peining

2016-01-01

Fluorescence in situ hybridization (FISH) is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes, and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains, and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbias and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH) allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH) using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells DNA and RNA FISH

Assessment of elimination profile of albendazole residues in fish.

PubMed

Busatto, Zenaís; de França, Welliton Gonçalves; Cyrino, José Eurico Possebon; Paschoal, Jonas Augusto Rizzato

2018-01-01

Few drugs are specifically regulated for aquaculture. Thus this study considered albendazole (ABZ) as a potential drug for use in fish, which, however, is not yet regulated for this application. ABZ is a broad-spectrum anthelmintic approved for farmed ruminants and recently considered for treatment of fish parasites. It is the subject of careful monitoring because of potential residues in animal products. This study evaluated the depletion of ABZ and its main known metabolites: albendazole sulfoxide - ABZSO, albendazole sulfone - ABZSO 2 and albendazole amino sulfone - ABZ-2-NH 2 SO 2 , in the fillets of the Neotropical Characin pacu, Piaractus mesopotamicus, which were fed diets containing 10 mg ABZ kg -1 body weight in a single dose. Fish were euthanised at 8, 12, 24, 48, 72, 96 and 120 hours after medication and the depletion profiles of ABZ, each metabolite and the sum of all marker residues were assessed and evaluated taking into account methodological variations regarding determination of the maximum residue limits adopted by different international regulating agencies for estimation of the withdrawal period (WP). The estimated WPs ranged from 2 to 7 days.

Hybrid vehicle potential assessment. Volume 7: Hybrid vehicle review

NASA Technical Reports Server (NTRS)

Leschly, K. O.

1979-01-01

Review of hybrid vehicles built during the past ten years or planned to be built in the near future is presented. An attempt is made to classify and analyze these vehicles to get an overall picture of their key characteristics. The review includes onroad hybrid passenger cars, trucks, vans, and buses.

FISH analysis for diagnostic evaluation of challenging melanocytic lesions.

PubMed

Zimmermann, A K; Hirschmann, A; Pfeiffer, D; Paredes, B E; Diebold, J

2010-09-01

The differential diagnosis of malignant melanomas and atypical melanocytic nevi is still a diagnostic challenge. The currently accepted morphologic criteria show substantial interobserver variability, likewise immunohistochemical studies are often not able to discriminate these lesions reliably. Techniques that support diagnostic accuracy are of the greatest importance considering the growing incidence of malignant melanomas and their increase in younger patients. In this study we analyzed the feasibility of fluorescence in situ hybridization (FISH) analysis for the discrimination of malignant and benign melanocytic tumors. A panel of DNA probes was used to detect chromosomal aberrations of chromosomes 6 and 11. On a series of 5 clearly malignant and benign melanocytic tumors we confirmed the applicability of the test. Then we focused on examination of ambiguous melanocytic lesions, where atypical cells are often difficult to relocalize in the 4',6-Diamidino-2-phenylindol (DAPI)-fluorescence stain. FISH analyses were conducted on destained H&E-stained slides. By comparison of the DAPI-image with photos taken from the H&E stain, unambiguous assignment of the FISH results to the conspicuous groups of cells was possible. The results of FISH analysis were consistent with the conventional diagnosis in 11 of 14 small ambiguous lesions. Of the remaining 3 cases, 2 showed FISH-results close to the cut-off level. Comparison of FISH results on thin and thick sections revealed that the cut-off values have to be adapted for 2 microm destained sections. In conclusion, FISH analysis is a useful and applicable tool for assessment of even smallest melanocytic neoplasms, although there will remain unclear cases that cannot be solved even after additional FISH evaluation.

Comparability of fish-based ecological quality assessments for geographically distinct Iberian regions.

PubMed

Segurado, P; Caiola, N; Pont, D; Oliveira, J M; Delaigue, O; Ferreira, M T

2014-04-01

In this work we compare two Iberian and a pan-European fish-based methods to assess ecological quality in rivers: the Fish-based Index of Biotic Integrity for Portuguese Wadeable Streams (F-IBIP), the Mediterranean Index of Biotic Integrity (IBIMED) and the pan-European Fish Index (EFI+). The results presented herein were developed in the context of the 2nd phase of the Intercalibration Exercise (IC), as required by the Water Frame Directive (WFD). The IC is aimed at ensuring comparability of the quality boundaries among the different WFD assessment methods developed by the Member States for each biological quality element. Although the two national assessment methods were developed for very distinct regions of Iberia (Western and Eastern Iberian Peninsula) they share the same methodological background: both are type-specific and guild-based multimetric indices. EFI+ is a multimetric guild-based model, but it is site-specific and uses a predictive modelling approach. The three indices were computed for all sites included in the Iberian Intercalibration database to allow the direct comparison, by means of linear regressions, of the resulting three quality values per site. The quality boundary harmonization between the two Iberian methods was only possible through an indirect comparison between the two indices, using EFI+ as a common metric. The three indices were also shown to be responsive to a common set of human induced pressures. This study highlights the need to develop general assessment methods adapted to wide geographical ranges with high species turnover to help intercalibrating assessment methods tailored for geographically more restricted regions. © 2013.

FISH and PNA FISH for the diagnosis of Q fever endocarditis and vascular infections.

PubMed

Prudent, Elsa; Lepidi, Hubert; Angelakis, Emmanouil; Raoult, Didier

2018-06-13

Purpose. Endocarditis and vascular infections are common manifestations of persistent localized infection due to Coxiella burnetii and recently, fluorescence in situ hybridization (FISH) was proposed as an alternative tool for their diagnosis. In this study, we evaluated the efficiency of FISH in a series of valve and vascular samples infected by C. burnetii. Methods. We tested 23 C. burnetii -positive valves and thrombus samples obtained from patients with Q fever endocarditis. Seven aneurysms and thrombus specimens were retrieved from patients with Q fever vascular infection. Samples were analyzed by culture, immunochemistry and FISH with oligonucleotide and PNA probes targeting C. burnetii -specific 16S ribosomal RNA sequences. Results. Immunohistochemical analysis was positive for five (17%) samples with significantly more copies of C. burnetii DNA than the negative ones ( p= 0.02). FISH was positive for 13 (43%) samples and presented 43% and 40% sensitivity compared to qPCR and culture, respectively. PNA FISH detected C. burnetii in 18 (60%) samples and presented 60% and 55% sensitivity compared to qPCR and culture, respectively. Immunohistochemistry had 38% and 28% sensitivity compared to FISH and PNA FISH, respectively. Samples found positive by both immunohistochemistry and PNA FISH contained significantly more copies of C. burnetii DNA than the negative ones ( p= 0.03). Finally, PNA FISH was more sensitive than FISH (60% versus 43%) for the detection of C. burnetii Conclusion. We provide evidence that PNA FISH and FISH are important assays for the diagnosis of C. burnetii endocarditis and vascular infections. Copyright © 2018 American Society for Microbiology.

2008-09 National Rivers and Streams Assessment Fish Tissue Data Dictionary

EPA Pesticide Factsheets

The Office of Science and Technology (OST) is providing the fish tissue results from the 2008-09 National Rivers and Streams Assessment (NRSA). This document includes the “data dictionary” for Mercury, Selenium, PBDEs, PCBs, Pesticides and PFCs.

Combining Telephone Surveys and Fishing Catches Self-Report: The French Sea Bass Recreational Fishery Assessment

PubMed Central

Rocklin, Delphine; Levrel, Harold; Drogou, Mickaël; Herfaut, Johanna; Veron, Gérard

2014-01-01

Fisheries statistics are known to be underestimated, since they are mainly based on information about commercial fisheries. However, various types of fishing activities exist and evaluating them is necessary for implementing effective management plans. This paper assesses the characteristics and catches of the French European sea bass recreational fishery along the Atlantic coasts, through the combination of large-scale telephone surveys and fishing diaries study. Our results demonstrated that half of the total catches (mainly small fish) were released at sea and that the mean length of a kept sea bass was 46.6 cm. We highlighted different patterns of fishing methods and type of gear used. Catches from boats were greater than from the shore, both in abundance and biomass, considering mean values per fishing trip as well as CPUE. Spearfishers caught the highest biomass of sea bass per fishing trip, but the fishing rod with lure was the most effective type of gear in terms of CPUE. Longlines had the highest CPUE value in abundance but not in biomass: they caught numerous but small sea bass. Handlines were less effective, catching few sea bass in both abundance and biomass. We estimated that the annual total recreational sea bass catches was 3,173 tonnes of which 2,345 tonnes were kept. Since the annual commercial catches landings were evaluated at 5,160 tonnes, recreational landings represent 30% of the total fishing catches on the Atlantic coasts of France. Using fishers' self-reports was a valuable way to obtain new information on data-poor fisheries. Our results underline the importance of evaluating recreational fishing as a part of the total amount of fisheries catches. More studies are critically needed to assess overall fish resources caught in order to develop effective fishery management tools. PMID:24489885

Combining telephone surveys and fishing catches self-report: the French sea bass recreational fishery assessment.

PubMed

Rocklin, Delphine; Levrel, Harold; Drogou, Mickaël; Herfaut, Johanna; Veron, Gérard

2014-01-01

Fisheries statistics are known to be underestimated, since they are mainly based on information about commercial fisheries. However, various types of fishing activities exist and evaluating them is necessary for implementing effective management plans. This paper assesses the characteristics and catches of the French European sea bass recreational fishery along the Atlantic coasts, through the combination of large-scale telephone surveys and fishing diaries study. Our results demonstrated that half of the total catches (mainly small fish) were released at sea and that the mean length of a kept sea bass was 46.6 cm. We highlighted different patterns of fishing methods and type of gear used. Catches from boats were greater than from the shore, both in abundance and biomass, considering mean values per fishing trip as well as CPUE. Spearfishers caught the highest biomass of sea bass per fishing trip, but the fishing rod with lure was the most effective type of gear in terms of CPUE. Longlines had the highest CPUE value in abundance but not in biomass: they caught numerous but small sea bass. Handlines were less effective, catching few sea bass in both abundance and biomass. We estimated that the annual total recreational sea bass catches was 3,173 tonnes of which 2,345 tonnes were kept. Since the annual commercial catches landings were evaluated at 5,160 tonnes, recreational landings represent 30% of the total fishing catches on the Atlantic coasts of France. Using fishers' self-reports was a valuable way to obtain new information on data-poor fisheries. Our results underline the importance of evaluating recreational fishing as a part of the total amount of fisheries catches. More studies are critically needed to assess overall fish resources caught in order to develop effective fishery management tools.

Global assessment of the status of coral reef herbivorous fishes: evidence for fishing effects.

PubMed

Edwards, C B; Friedlander, A M; Green, A G; Hardt, M J; Sala, E; Sweatman, H P; Williams, I D; Zgliczynski, B; Sandin, S A; Smith, J E

2014-01-07

On coral reefs, herbivorous fishes consume benthic primary producers and regulate competition between fleshy algae and reef-building corals. Many of these species are also important fishery targets, yet little is known about their global status. Using a large-scale synthesis of peer-reviewed and unpublished data, we examine variability in abundance and biomass of herbivorous reef fishes and explore evidence for fishing impacts globally and within regions. We show that biomass is more than twice as high in locations not accessible to fisheries relative to fisheries-accessible locations. Although there are large biogeographic differences in total biomass, the effects of fishing are consistent in nearly all regions. We also show that exposure to fishing alters the structure of the herbivore community by disproportionately reducing biomass of large-bodied functional groups (scraper/excavators, browsers, grazer/detritivores), while increasing biomass and abundance of territorial algal-farming damselfishes (Pomacentridae). The browser functional group that consumes macroalgae and can help to prevent coral-macroalgal phase shifts appears to be most susceptible to fishing. This fishing down the herbivore guild probably alters the effectiveness of these fishes in regulating algal abundance on reefs. Finally, data from remote and unfished locations provide important baselines for setting management and conservation targets for this important group of fishes.

Designing a global assessment of climate change on inland fishes and fisheries: knowns and needs

USGS Publications Warehouse

Paukert, Craig P.; Lynch, Abigail J.; Beard, T. Douglas; Chen, Yushun; Cooke, Steven J.; Cooperman, Michael S.; Cowx, Ian G.; Infante, Dana M.; Ibengwe, Lilian; Myers, Bonnie; Nguyen, Phu Hoa; Winfield, Ian J.

2017-01-01

To date, there are few comprehensive assessments of how climate change affects inland finfish, fisheries, and aquaculture at a global scale, but one is necessary to identify research needs and commonalities across regions and to help guide decision making and funding priorities. Broadly, the consequences of climate change on inland fishes will impact global food security, the livelihoods of people who depend on inland capture and recreational fisheries. However, understanding how climate change will affect inland fishes and fisheries has lagged behind marine assessments. Building from a North American inland fisheries assessment, we convened an expert panel from seven countries to provide a first-step to a framework for determining how to approach an assessment of how climate change may affect inland fishes, capture fisheries, and aquaculture globally. Starting with the small group helped frame the key questions (e.g., who is the audience? What is the best approach and spatial scale?). Data gaps identified by the group include: the tolerances of inland fisheries to changes in temperature, stream flows, salinity, and other environmental factors linked to climate change, and the adaptive capacity of fishes and fisheries to adjust to these changes. These questions are difficult to address, but long-term and large-scale datasets are becoming more readily available as a means to test hypotheses related to climate change. We hope this perspective will help researchers and decision makers identify research priorities and provide a framework to help sustain inland fish populations and fisheries for the diversity of users around the globe.

Temperature effect on gastric emptying time of hybrid grouper (Epinephelus spp.)

NASA Astrophysics Data System (ADS)

De, Moumita; Ghaffar, Mazlan Abd.; Das, Simon K.

2014-09-01

Knowledge of fish gastric emptying time is a necessary component for understanding the fish feeding rates, energy budgets and commercial production of fishes in aquaculture. The hybrid grouper Epinephelus spp. is getting popular as a culture species in Malaysia for their faster growth rate compared to commonly cultured grouper species (giant grouper Epinephelus lanceolatus and tiger grouper Epinephelus fuscoguttatus). There are data suggests that elevated sea water temperature affects gastric emptying time (GET) of fishes. Hence, this study aims to study the GET of hybrid grouper at different temperature (22, 26, 30, 34°C) in laboratory condition with commercial diet pellet. The gastric emptying times (GETs) at different temperatures were determined X-radiographically, using barium sulfate (BaSO4) as a contrast medium food marker. The food marker and X-radiography showed that initial voidance of fecal matter began 4-6 h after feeding at all temperature. The fastest GET (13 h) was obsereved in the 30°C group, whereas the longest (17 h) GET was seen in 22°C group fed with artificial diet pellet. Not much differences in GET were recorded between the 26 and 34°C groups as 34°C groups fed lesser amount compared to 26°C groups. Nevertheless a substantial delay in GET was observed in the 22°C group. The findings of this study suggest to culture hybrid grouper between 26 to 30°C with commercial diet pellet as this temperature ranges proliferate the faster digestion process which may contribute faster growth rate of this commerical important fish species. Overall, these findings may have important consequences for optimization of commercial production of hybrid grouper.

Evolutionary impact assessment: accounting for evolutionary consequences of fishing in an ecosystem approach to fisheries management

PubMed Central

Laugen, Ane T; Engelhard, Georg H; Whitlock, Rebecca; Arlinghaus, Robert; Dankel, Dorothy J; Dunlop, Erin S; Eikeset, Anne M; Enberg, Katja; Jørgensen, Christian; Matsumura, Shuichi; Nusslé, Sébastien; Urbach, Davnah; Baulier, Loїc; Boukal, David S; Ernande, Bruno; Johnston, Fiona D; Mollet, Fabian; Pardoe, Heidi; Therkildsen, Nina O; Uusi-Heikkilä, Silva; Vainikka, Anssi; Heino, Mikko; Rijnsdorp, Adriaan D; Dieckmann, Ulf

2014-01-01

Managing fisheries resources to maintain healthy ecosystems is one of the main goals of the ecosystem approach to fisheries (EAF). While a number of international treaties call for the implementation of EAF, there are still gaps in the underlying methodology. One aspect that has received substantial scientific attention recently is fisheries-induced evolution (FIE). Increasing evidence indicates that intensive fishing has the potential to exert strong directional selection on life-history traits, behaviour, physiology, and morphology of exploited fish. Of particular concern is that reversing evolutionary responses to fishing can be much more difficult than reversing demographic or phenotypically plastic responses. Furthermore, like climate change, multiple agents cause FIE, with effects accumulating over time. Consequently, FIE may alter the utility derived from fish stocks, which in turn can modify the monetary value living aquatic resources provide to society. Quantifying and predicting the evolutionary effects of fishing is therefore important for both ecological and economic reasons. An important reason this is not happening is the lack of an appropriate assessment framework. We therefore describe the evolutionary impact assessment (EvoIA) as a structured approach for assessing the evolutionary consequences of fishing and evaluating the predicted evolutionary outcomes of alternative management options. EvoIA can contribute to EAF by clarifying how evolution may alter stock properties and ecological relations, support the precautionary approach to fisheries management by addressing a previously overlooked source of uncertainty and risk, and thus contribute to sustainable fisheries. PMID:26430388

Evolutionary impact assessment: accounting for evolutionary consequences of fishing in an ecosystem approach to fisheries management.

PubMed

Laugen, Ane T; Engelhard, Georg H; Whitlock, Rebecca; Arlinghaus, Robert; Dankel, Dorothy J; Dunlop, Erin S; Eikeset, Anne M; Enberg, Katja; Jørgensen, Christian; Matsumura, Shuichi; Nusslé, Sébastien; Urbach, Davnah; Baulier, Loїc; Boukal, David S; Ernande, Bruno; Johnston, Fiona D; Mollet, Fabian; Pardoe, Heidi; Therkildsen, Nina O; Uusi-Heikkilä, Silva; Vainikka, Anssi; Heino, Mikko; Rijnsdorp, Adriaan D; Dieckmann, Ulf

2014-03-01

Managing fisheries resources to maintain healthy ecosystems is one of the main goals of the ecosystem approach to fisheries (EAF). While a number of international treaties call for the implementation of EAF, there are still gaps in the underlying methodology. One aspect that has received substantial scientific attention recently is fisheries-induced evolution (FIE). Increasing evidence indicates that intensive fishing has the potential to exert strong directional selection on life-history traits, behaviour, physiology, and morphology of exploited fish. Of particular concern is that reversing evolutionary responses to fishing can be much more difficult than reversing demographic or phenotypically plastic responses. Furthermore, like climate change, multiple agents cause FIE, with effects accumulating over time. Consequently, FIE may alter the utility derived from fish stocks, which in turn can modify the monetary value living aquatic resources provide to society. Quantifying and predicting the evolutionary effects of fishing is therefore important for both ecological and economic reasons. An important reason this is not happening is the lack of an appropriate assessment framework. We therefore describe the evolutionary impact assessment (EvoIA) as a structured approach for assessing the evolutionary consequences of fishing and evaluating the predicted evolutionary outcomes of alternative management options. EvoIA can contribute to EAF by clarifying how evolution may alter stock properties and ecological relations, support the precautionary approach to fisheries management by addressing a previously overlooked source of uncertainty and risk, and thus contribute to sustainable fisheries.

Assessment of ERBB2 and EGFR gene amplification and protein expression in gastric carcinoma by immunohistochemistry and fluorescence in situ hybridization

PubMed Central

2011-01-01

Background The goal of this study was to investigate ERBB2(HER2) and EGFR gene amplification and protein expression in gastric cancer. Fluorescence in situ hybridization (FISH) and immunohistochemistry were used to analyze ERBB2 and EGFR gene amplification and protein expression in 69 cases of gastric cancer. Results FISH analysis revealed that 20.3% of the cases exhibited ERBB2 gene amplification. Increases in ERBB2 copy number and gene amplification were present in 52.2% of the samples. Expression of the ERBB2 protein was observed in 42.0% of cases. FISH analysis detected EGFR gene amplification in 29.0% of samples. Increases in EGFR copy number and gene amplification occurred in 57.9% of samples, and EGFR protein expression was present in 52.2% of samples. Both ERBB2 and EGFR gene amplification were 3 cases (4.3%), but abnormalities in both ERBB2 and EGFR gene copy number were present 36.2% of samples. ERBB2 and EGFR gene amplification were significantly associated with the depth of tumor invasion (P < 0.05) and lymph node metastasis (P < 0.05), but not with sex, age, or histological type (P > 0.05). Conclusions Our data indicated that ERBB2 and EGFR genetic abnormalities were associated with the prognosis of gastric cancer. Clinical assessment of ERBB2 and EGFR amplification may represent an important factor for the development of personalized treatment programs for gastic cancer. PMID:21689422

Detection and signal amplification in zebrafish RNA FISH.

PubMed

Hauptmann, Giselbert; Lauter, Gilbert; Söll, Iris

2016-04-01

In situ hybridization (ISH) has become an invaluable tool for the detection of RNA in cells, tissues and organisms. Due to improvements in target and signal amplification and in probe design remarkable progress has been made concerning sensitivity, specificity and resolution of chromogenic and fluorescent ISH (FISH). These advancements allow for exquisite cellular and sub-cellular resolution and for detecting multiple RNA species at a time by multiplexing. In zebrafish (F)ISH non-enzymatic and enzymatic amplification systems have been employed to obtain enhanced signal intensities and signal-to-noise ratios. These amplification strategies include branched DNA-based RNAscope and in situ hybridization chain reaction (HCR) techniques, as well as alkaline phosphatase (AP)- and horseradish peroxidase (PO)-based immunoassays. For practical application, we provide proven multiplex FISH protocols for AP- and PO-based visualization of mRNAs at high resolution. The protocols take advantage of optimized tyramide signal amplification (TSA) conditions of the PO assay and long-lasting high signal-to-noise ratio of the AP reaction, thereby enabling detection of less abundant transcripts. Copyright © 2016 Elsevier Inc. All rights reserved.

Uterine cavity lavage: adding FISH to conventional cytogenetics for embryonic sexing and diagnosing common chromosomal aberrations.

PubMed

Ishai, D; Amiel, A; Diukman, R; Cogan, O; Lichtenstein, Z; Abramovici, H; Fejgin, M D

1995-10-01

This study was undertaken to examine the efficacy for early prenatal diagnosis of uterine cavity lavage at the level of the internal os and to assess the rate of maternal contamination. In phase I, uterine cavity lavage was performed in 38 women scheduled for pregnancy termination between 6 and 12 weeks. In addition to short- and long-term cultures, one-colour FISH (fluorescence in situ hybridization) with Y and X probes was used for fetal sexing. Two-colour FISH was used in all known male fetuses for the assessment of maternal contamination. In phase II, lavage was performed on 16 women. Fetal sex was diagnosed with direct labelled X and Y probes and common numerical chromosomal aberration was attempted with 18 and 21 direct labelled probes. Fetal sexing was successful in all cases in phases I and II. Out of 34 patients in which tissue was obtained, only FISH was done in six. Long-term cell cultures were successful in the other 28 cases, but complete karyotyping in 19 (56 per cent). No chromosomal aberration was found with the direct labelled probes 18 and 21 in FISH. Maternal contamination was assessed to be 5-10 per cent. This simple and easy-to-master technique is very effective in obtaining fetal cells early in pregnancy for genetic diagnosis, especially by FISH. However, the safety of the procedure must be tested in ongoing pregnancies.

Spatial segregation of spawning habitat limits hybridization between sympatric native Steelhead and Coastal Cutthroat Trout

USGS Publications Warehouse

Buehrens, T.W.; Glasgow, J.; Ostberg, Carl O.; Quinn, T.P.

2013-01-01

Native Coastal Cutthroat Trout Oncorhynchus clarkii clarkii and Coastal Steelhead O. mykiss irideus hybridize naturally in watersheds of the Pacific Northwest yet maintain species integrity. Partial reproductive isolation due to differences in spawning habitat may limit hybridization between these species, but this process is poorly understood. We used a riverscape approach to determine the spatial distribution of spawning habitats used by native Coastal Cutthroat Trout and Steelhead as evidenced by the distribution of recently emerged fry. Molecular genetic markers were used to classify individuals as pure species or hybrids, and individuals were assigned to age-classes based on length. Fish and physical habitat data were collected in a spatially continuous framework to assess the relationship between habitat and watershed features and the spatial distribution of parental species and hybrids. Sampling occurred in 35 reaches from tidewaters to headwaters in a small (20 km2) coastal watershed in Washington State. Cutthroat, Steelhead, and hybrid trout accounted for 35%, 42%, and 23% of the fish collected, respectively. Strong segregation of spawning areas between Coastal Cutthroat Trout and Steelhead was evidenced by the distribution of age-0 trout. Cutthroat Trout were located farther upstream and in smaller tributaries than Steelhead were. The best predictor of species occurrence at a site was the drainage area of the watershed that contributed to the site. This area was positively correlated with the occurrence of age-0 Steelhead and negatively with the presence of Cutthroat Trout, whereas hybrids were found in areas occupied by both parental species. A similar pattern was observed in older juveniles of both species but overlap was greater, suggesting substantial dispersal of trout after emergence. Our results offer support for spatial reproductive segregation as a factor limiting hybridization between Steelhead and Coastal Cutthroat Trout.

Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens.

PubMed

García-Caballero, Tomás; Grabau, Dorthe; Green, Andrew R; Gregory, John; Schad, Arno; Kohlwes, Elke; Ellis, Ian O; Watts, Sarah; Mollerup, Jens

2010-03-01

Fluorescence in situ hybridization (FISH) can be used to reveal several genomic imbalances relevant to proper cancer diagnosis and to the correct treatment regime. However, FISH requires expensive and advanced fluorescence microscopes in addition to expertise in fluorescence microscopy. To determine whether a newly developed dual-colour chromogenic in situ hybridization (CISH) method is a suitable alternative to FISH, we analysed the human epidermal growth factor receptor 2 gene (HER2) amplification level of 168 breast cancer specimens using dual-colour CISH and FISH and compared the results. We found 100% agreement between HER2 status determined by FISH and dual-colour CISH. Furthermore, we observed that the time used to score slides was significantly reduced by 28% in dual-colour CISH compared with the FISH protocol. Concordance between HER2 protein status and dual-colour CISH or FISH was equally good with an overall agreement of 96.8%. Correlation between the HER2/centromere 17 gene ratios obtained with dual-colour CISH and FISH was highly significant with an overall correlation coefficient (rho) of 0.96. We conclude that dual-colour CISH and bright field microscopy are excellent alternatives to FISH when analysing the HER2 status of primary breast cancer.

Clinical methods for the assessment of the effects of environmental stress on fish health

USGS Publications Warehouse

Wedemeyer, Gary A.; Yasutake, William T.

1977-01-01

Clinical methods are presented for biological monitoring of hatchery and native fish populations to assess the effects of environmental stress on fish health. The choice of methods is based on the experience of the authors and the judgment of colleagues at fishery laboratories of the U.S. Fish and Wildlife Service. Detailed analysis methods, together with guidelines for sample collection and for the interpretation of results, are given for tests on blood (cell counts, chloride, cholesterol, clotting time, cortisol, glucose, hematocrit, hemoglobin, lactic acid, methemoglobin, osmolality, and total protein); water (ammonia and nitrite content); and liver and muscle (glycogen content).

CONTAMINATION OF FISH IN STREAMS OF THE MID-ATLANTIC REGION: AN APPROACH TO REGIONAL INDICATOR SELECTION AND WILDLIFE ASSESSMENT

EPA Science Inventory

The extent of contamination of fish in the Mid-Atlantic Region was evaluated as part of the U.S. Environmental Protection Agency's Monitoring and Assessment Program's regional assessment in 1993 through 1994. Fish assemblages from wadeable streams were dominated by small, short-...

Nutrient enrichment and fish nutrient tolerance: Assessing biologically relevant nutrient criteria

USGS Publications Warehouse

Meador, Michael R.

2013-01-01

Relationships between nutrient concentrations and fish nutrient tolerance were assessed relative to established nutrient criteria. Fish community, nitrate plus nitrite (nitrate), and total phosphorus (TP) data were collected during summer low-flow periods in 2003 and 2004 at stream sites along a nutrient-enrichment gradient in an agricultural basin in Indiana and Ohio and an urban basin in the Atlanta, Georgia, area. Tolerance indicator values for nitrate and TP were assigned for each species and averaged separately for fish communities at each site (TIVo). Models were used to predict fish species expected to occur at a site under minimally disturbed conditions and average tolerance indicator values were determined for nitrate and TP separately for expected communities (TIVe). In both areas, tolerance scores (TIVo/TIVe) for nitrate increased significantly with increased nitrate concentrations whereas no significant relationships were detected between TP tolerance scores and TP concentrations. A 0% increase in the tolerance score (TIVo/TIVe = 1) for nitrate corresponded to a nitrate concentration of 0.19 mg/l (compared with a USEPA summer nitrate criterion of 0.17 mg/l) in the urban area and 0.31 mg/l (compared with a USEPA summer nitrate criterion of 0.86 mg/l) in the agricultural area. Fish nutrient tolerance values offer the ability to evaluate nutrient enrichment based on a quantitative approach that can provide insights into biologically relevant nutrient criteria.

Temperature effect on gastric emptying time of hybrid grouper (Epinephelus spp.)

DOE Office of Scientific and Technical Information (OSTI.GOV)

De, Moumita; Ghaffar, Mazlan Abd.; Das, Simon K.

2014-09-03

Knowledge of fish gastric emptying time is a necessary component for understanding the fish feeding rates, energy budgets and commercial production of fishes in aquaculture. The hybrid grouper Epinephelus spp. is getting popular as a culture species in Malaysia for their faster growth rate compared to commonly cultured grouper species (giant grouper Epinephelus lanceolatus and tiger grouper Epinephelus fuscoguttatus). There are data suggests that elevated sea water temperature affects gastric emptying time (GET) of fishes. Hence, this study aims to study the GET of hybrid grouper at different temperature (22, 26, 30, 34°C) in laboratory condition with commercial diet pellet.more » The gastric emptying times (GETs) at different temperatures were determined X-radiographically, using barium sulfate (BaSO{sub 4}) as a contrast medium food marker. The food marker and X-radiography showed that initial voidance of fecal matter began 4-6 h after feeding at all temperature. The fastest GET (13 h) was obsereved in the 30°C group, whereas the longest (17 h) GET was seen in 22°C group fed with artificial diet pellet. Not much differences in GET were recorded between the 26 and 34°C groups as 34°C groups fed lesser amount compared to 26°C groups. Nevertheless a substantial delay in GET was observed in the 22°C group. The findings of this study suggest to culture hybrid grouper between 26 to 30°C with commercial diet pellet as this temperature ranges proliferate the faster digestion process which may contribute faster growth rate of this commerical important fish species. Overall, these findings may have important consequences for optimization of commercial production of hybrid grouper.« less

Assessment of hepatic gene expression between hybrid striped bass exhibiting extremes in growth performance

USDA-ARS?s Scientific Manuscript database

Hybrid striped bass is a major aquaculture species in the United States. Artificial breeding of this species can introduce large variation in growth performance during grow-out to market size. To assess the genetic and nutrigenomic basis behind growth variability in these hybrids, fingerlings from 4...

A General Model of Distant Hybridization Reveals the Conditions for Extinction in Atlantic Salmon and Brown Trout

PubMed Central

Quilodrán, Claudio S.; Currat, Mathias; Montoya-Burgos, Juan I.

2014-01-01

Interspecific hybridization is common in nature but can be increased in frequency or even originated by human actions, such as species introduction or habitat modification, which may threaten species persistence. When hybridization occurs between distantly related species, referred to as “distant hybridization,” the resulting hybrids are generally infertile or fertile but do not undergo chromosomal recombination during gametogenesis. Here, we present a model describing this frequent but poorly studied interspecific hybridization to assess its consequences on parental species and to anticipate the conditions under which they can reach extinction. Our general model fully incorporates three important processes: density-dependent competition, dominance/recessivity inheritance of traits and assortative mating. We demonstrate its use and flexibility by assessing population extinction risk between Atlantic salmon and brown trout in Norway, whose interbreeding has recently increased due to farmed fish releases into the wild. We identified the set of conditions under which hybridization may threaten salmonid species. Thanks to the flexibility of our model, we evaluated the effect of an additional risk factor, a parasitic disease, and showed that the cumulative effects dramatically increase the extinction risk. The consequences of distant hybridization are not genetically, but demographically mediated. Our general model is useful to better comprehend the evolution of such hybrid systems and we demonstrated its importance in the field of conservation biology to set up management recommendations when this increasingly frequent type of hybridization is in action. PMID:25003336

A general model of distant hybridization reveals the conditions for extinction in Atlantic salmon and brown trout.

PubMed

Quilodrán, Claudio S; Currat, Mathias; Montoya-Burgos, Juan I

2014-01-01

Interspecific hybridization is common in nature but can be increased in frequency or even originated by human actions, such as species introduction or habitat modification, which may threaten species persistence. When hybridization occurs between distantly related species, referred to as "distant hybridization," the resulting hybrids are generally infertile or fertile but do not undergo chromosomal recombination during gametogenesis. Here, we present a model describing this frequent but poorly studied interspecific hybridization to assess its consequences on parental species and to anticipate the conditions under which they can reach extinction. Our general model fully incorporates three important processes: density-dependent competition, dominance/recessivity inheritance of traits and assortative mating. We demonstrate its use and flexibility by assessing population extinction risk between Atlantic salmon and brown trout in Norway, whose interbreeding has recently increased due to farmed fish releases into the wild. We identified the set of conditions under which hybridization may threaten salmonid species. Thanks to the flexibility of our model, we evaluated the effect of an additional risk factor, a parasitic disease, and showed that the cumulative effects dramatically increase the extinction risk. The consequences of distant hybridization are not genetically, but demographically mediated. Our general model is useful to better comprehend the evolution of such hybrid systems and we demonstrated its importance in the field of conservation biology to set up management recommendations when this increasingly frequent type of hybridization is in action.

Microbiological assessment along the fish production chain of the Norwegian pelagic fisheries sector--Results from a spot sampling programme.

PubMed

Svanevik, Cecilie Smith; Roiha, Irja Sunde; Levsen, Arne; Lunestad, Bjørn Tore

2015-10-01

Microbes play an important role in the degradation of fish products, thus better knowledge of the microbiological conditions throughout the fish production chain may help to optimise product quality and resource utilisation. This paper presents the results of a ten-year spot sampling programme (2005-2014) of the commercially most important pelagic fish species harvested in Norway. Fish-, surface-, and storage water samples were collected from fishing vessels and processing factories. Totally 1,181 samples were assessed with respect to microbiological quality, hygiene and food safety. We introduce a quality and safety assessment scheme for fresh pelagic fish recommending limits for heterotrophic plate counts (HPC), thermos tolerant coliforms, enterococci and Listeria monocytogenes. According to the scheme, in 25 of 41 samplings, sub-optimal conditions were found with respect to quality, whereas in 21 and 9 samplings, samples were not in compliance concerning hygiene and food safety, respectively. The present study has revealed that the quality of pelagic fish can be optimised by improving the hygiene conditions at some critical points at an early phase of the production chain. Thus, the proposed assessment scheme may provide a useful tool for the industry to optimise quality and maintain consumer safety of pelagic fishery products. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

The role of hybridization in the distribution, conservation and management of aquatic species: Symposium review

USGS Publications Warehouse

Epifanio, John; Nielsen, Jennifer L.

2001-01-01

This issue of Reviews in Fish Biology and Fisheries contains six papers addressing several critical aspects of hybridization in fishes and aquatic organisms. Hybridization is a phenomenon long recognized in fishes (Hubbs, 1920, 1955; Schwarz, 1981), as well as in other plant and vertebrate taxa, despite some rather dogmatic proclamations to the contrary, e.g., comments made by David Starr Jordan at the beginning of the 20th century that the species “line” is rarely crossed in fishes (Clark Hubbs, personal communication). Since that time, interspecific genetic introgression has been well documented in many fish genera and species: Barbus (Berrebi and CattaneoBerrebi, 1993); Cyprinodon (Echelle and Connor, 1989; Dowling and DeMarais, 1993); Gambusia (Hubbs, 1959; Scribner and Avise, 1994); Esox (Wahl and Stein, 1993); Lepomis (Avise et al., 1984); Luxilus (Duvernell and Aspinwall, 1995); Morone (Harrell et al., 1993); Notropis (Dowling et al., 1989; Dowling and Hoeh, 1991); Oncorhynchus (Busack and Gall, 1981; Campton and Utter, 1985; Loudenslager et al., 1986; Leary et al., 1987; Forbes and Allendorf, 1991; Dowling and Childs, 1992); Salmo (Nyman, 1970; Wilkins et al., 1993; Giuffra et al., 1996; Hartley, 1996; Perez et al., 1999); Salvalinus (Hammar et al., 1991; Bernatchez et al., 1995; Baxter et al., 1997; Glemet et al., 1998; Wilson and Bernatchez, 1998); Sebastes (Seeb, 1988); Stizostedion (Billington et al., 1988). See also reviews in Campton (1987), Verspoor and Hammar (1991), Smith (1992), and Scribner et al. (2000). More recently, a number of investigations have documented not only first generation hybrids, but also subsequent generation introgressant hybrids (Bartley et al., 1990; Verspoor and Hammar, 1991). As a result, our views about species typology and hybrids continue to change.

Discriminating multi-species populations in biofilms with peptide nucleic acid fluorescence in situ hybridization (PNA FISH).

PubMed

Almeida, Carina; Azevedo, Nuno F; Santos, Sílvio; Keevil, Charles W; Vieira, Maria J

2011-03-29

Our current understanding of biofilms indicates that these structures are typically composed of many different microbial species. However, the lack of reliable techniques for the discrimination of each population has meant that studies focusing on multi-species biofilms are scarce and typically generate qualitative rather than quantitative data. We employ peptide nucleic acid fluorescence in situ hybridization (PNA FISH) methods to quantify and visualize mixed biofilm populations. As a case study, we present the characterization of Salmonella enterica/Listeria monocytogenes/Escherichia coli single, dual and tri-species biofilms in seven different support materials. Ex-situ, we were able to monitor quantitatively the populations of ∼56 mixed species biofilms up to 48 h, regardless of the support material. In situ, a correct quantification remained more elusive, but a qualitative understanding of biofilm structure and composition is clearly possible by confocal laser scanning microscopy (CLSM) at least up to 192 h. Combining the data obtained from PNA FISH/CLSM with data from other established techniques and from calculated microbial parameters, we were able to develop a model for this tri-species biofilm. The higher growth rate and exopolymer production ability of E. coli probably led this microorganism to outcompete the other two [average cell numbers (cells/cm(2)) for 48 h biofilm: E. coli 2,1 × 10(8) (± 2,4 × 10(7)); L. monocytogenes 6,8 × 10(7) (± 9,4 × 10(6)); and S. enterica 1,4 × 10(6) (± 4,1 × 10(5))]. This overgrowth was confirmed by CSLM, with two well-defined layers being easily identified: the top one with E. coli, and the bottom one with mixed regions of L. monocytogenes and S. enterica. While PNA FISH has been described previously for the qualitative study of biofilm populations, the present investigation demonstrates that it can also be used for the accurate quantification and spatial distribution of species in polymicrobial communities

FISH in polycythemia vera (PCV)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Amiel, A.; Gaber, E.; Manor, Y.

1994-09-01

Trisomies 8 and 9 are the most common numerical abnormalities in polycythemia vera (PCV). However, their role in the pathogenesis of the disease is unclear as is their diagnostic or prognostic value. We evaluated the role of fluorescent in-situ hybridization (FISH) as compared to chromosome analysis in the detection of trisomies 8 or 9 in peripheral blood cells of 14 PCV and 5 secondary PCV patients. Using FISH, we found trisomies 8 and 9 in 10 PCV patients above the cutoff levels of 5%. However, no patient with the secondary PCV reached the cutoff level. Out of 10 PCV patientsmore » in whom the trisomy was detected by FISH, only in 3 was this trisomy also detected by routine cytogenetics. The incidence of the finding of trisomy 9 correlates with the duration of the disease, suggesting that this is not the primary event in PCV. FISH is a sensitive, convenient and rapid method for diagnosis and follow-up of chromosome aberrations in PCV patients. Application of FISH to larger cohort of patients may provide valuable information regarding their role in initiation and progession of the disease.« less

ALK status testing in non-small cell lung carcinoma: correlation between ultrasensitive IHC and FISH.

PubMed

Minca, Eugen C; Portier, Bryce P; Wang, Zhen; Lanigan, Christopher; Farver, Carol F; Feng, Yan; Ma, Patrick C; Arrossi, Valeria A; Pennell, Nathan A; Tubbs, Raymond R

2013-05-01

ALK gene rearrangements in advanced non-small cell lung carcinomas (NSCLC) are an indication for targeted therapy with crizotinib. Fluorescence in situ hybridization (FISH) using a recently approved companion in vitro diagnostic class FISH system commonly assesses ALK status. More accessible IHC is challenged by low expression of ALK-fusion transcripts in NSCLC. We compared ultrasensitive automated IHC with FISH for detecting ALK status on 318 FFPE and 40 matched ThinPrep specimens from 296 patients with advanced NSCLC. IHC was concordant with FFPE-FISH on 229 of 231 dual-informative samples (31 positive and 198 negative) and with ThinPrep-FISH on 34 of 34 samples (5 positive and 29 negative). Two cases with negative IHC and borderline-positive FFPE-FISH (15% and 18%, respectively) were reclassified as concordant based on negative matched ThinPrep-FISH and clinical data consistent with ALK-negative status. Overall, after including ThinPrep-FISH and amending the false-positive FFPE-FISH results, IHC demonstrated 100% sensitivity and specificity (95% CI, 0.86 to 1.00 and 0.97 to 1.00, respectively) for ALK detection on 249 dual-informative NSCLC samples. IHC was informative on significantly more samples than FFPE-FISH, revealing additional ALK-positive cases. The high concordance with FISH warrants IHC's routine use as the initial component of an algorithmic approach to clinical ALK testing in NSCLC, followed by reflex FISH confirmation of IHC-positive cases. Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Cytonuclear genetics of experimental fish hybrid zones inside Biosphere 2

USGS Publications Warehouse

Scribner, K.T.; Avise, John C.

1994-01-01

Two species of mosquitofish (family Poeciliidae) known to hybridize in nature were introduced into freshwater habitats inside Biosphere 2, and their population genetics were monitored after 2 years. Within four to six generations, nuclear and cytoplasmic markers characteristic of Gambusia holbrooki had risen greatly in frequency, although some Gambusia affinis alleles and haplotypes were retained primarily in recombinant genotypes, indicative of introgressive hybridization. The temporal cytonuclear dynamics proved similar to population genetic changes observed in replicated experimental hybrid populations outside of Biosphere 2, thus indicating strong directional selection favoring G. holbrooki genotypes across the range of environments monitored. When interpreted in the context of species-specific population demographies observed previously, results suggest that the extremely rapid evolution in these zones of secondary contact is attributable primarily to species differences in life-history traits.

Hybrid mechanosensing system to generate the polarity needed for migration in fish keratocytes

PubMed Central

Okimura, Chika; Iwadate, Yoshiaki

2016-01-01

ABSTRACT Crawling cells can generate polarity for migration in response to forces applied from the substratum. Such reaction varies according to cell type: there are both fast- and slow-crawling cells. In response to periodic stretching of the elastic substratum, the intracellular stress fibers in slow-crawling cells, such as fibroblasts, rearrange themselves perpendicular to the direction of stretching, with the result that the shape of the cells extends in that direction; whereas fast-crawling cells, such as neutrophil-like differentiated HL-60 cells and Dictyostelium cells, which have no stress fibers, migrate perpendicular to the stretching direction. Fish epidermal keratocytes are another type of fast-crawling cell. However, they have stress fibers in the cell body, which gives them a typical slow-crawling cell structure. In response to periodic stretching of the elastic substratum, intact keratocytes rearrange their stress fibers perpendicular to the direction of stretching in the same way as fibroblasts and migrate parallel to the stretching direction, while blebbistatin-treated stress fiber-less keratocytes migrate perpendicular to the stretching direction, in the same way as seen in HL-60 cells and Dictyostelium cells. Our results indicate that keratocytes have a hybrid mechanosensing system that comprises elements of both fast- and slow-crawling cells, to generate the polarity needed for migration. PMID:27124267

Effects of dietary green tea (Camellia sinensis L.) supplementation on growth performance, lipid metabolism, and antioxidant status in a sturgeon hybrid of Sterlet (Huso huso ♂ × Acipenser ruthenus ♀) fed oxidized fish oil.

PubMed

Hasanpour, Soleiman; Salati, Amir Parviz; Falahatkar, Bahram; Azarm, Hamid Mohammadi

2017-10-01

Lipid content of diet is very susceptible to oxidation, especially when stored for a long time, so for evaluating protective effects of green tea in fish received oxidized oil, this study was done. Lipid content of diet was replaced by oxidized fish oil (OFO) in 0, 50, and 100%. Green tea extract (GTE) was added to diet in three levels, 0, 5, and 100 mg/kg giving a total of nine experimental diets. Two hundred and seventy sturgeon hybrid of Sterlet (Huso huso ♀ × Acipenser ruthenus ♂) with initial weight of 212.6 ± 0.7 g after 2 weeks adaptation randomly divided in 27 fiberglass tanks with 700 L volume. Fish were fed satiated three times daily. After 6 weeks, biometry was done to evaluate growth performance and blood samples were taken for biochemical analysis. The result showed that feeding with oxidized oil had no effects on growth. However, in fish fed GTE, growth indices improved slightly. Feeding with OFO reduced serum total cholesterol, triacylglycerol, and low-density lipoprotein, while increased high density lipoprotein. Dietary GTE moderated the effects of OFO on lipid metabolism. Feeding with the OFO increased activity of serum superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde contents. In fish received both OFO and GTE, reduced activity of serum antioxidant enzymes and malondialdehyde content was recorded in compare to fish fed only OFO. According to the result of the present study, it can be argued that feeding of sturgeon hybrid of Sterlet with OFO has negative effects on lipid metabolism and antioxidant status, whereas GTE dosages used in this study have protective effects on fish from the adverse effects of oxidized oil.

Effect of grading fingerling hybrid catfish (Channel Catfish X Blue Catfish) on growth, production, feed conversion, and foodfish size distribution.

USDA-ARS?s Scientific Manuscript database

Hybrid catfish production ponds often produce a wide size range of fish and payments to farmers may be reduced due to discounts for larger fish. This study was conducted to determine effect of grading hybrid catfish fingerlings on the size distribution of harvested foodfish. Three 0.25-acre ponds we...

Hybrid Vehicle Technology Constraints and Application Assessment Study : Volume 1. Summary.

DOT National Transportation Integrated Search

1977-11-01

This four-volume report presents analyses and assessments of both heat engine/battery- and heat engine/flywheel-powered hybrid vehicles to determine if they could contribute to near-term (1980-1990) reductions in transportation energy consumption und...

Fishes, mussels, crayfishes, and aquatic habitats of the Hoosier-Shawnee ecological assessment area

Treesearch

M. Burr Brooks; Justin T. Sipiorski; Matthew R. Thomas; Kevin S. Cummings; Christopher A. Taylor

2004-01-01

The Hoosier-Shawnee Ecological Assessment Area, part of the Coastal Plain and Interior Low Plateau physiographic provinces, includes 194 native fish species, 76 native mussel species, and 34 native crayfish species. Five of the subregions (e.g., Mississippi Embayment) that make up the assessment area were recently ranked as either globally or bioregionally outstanding...

CARD-FISH for Environmental Microorganisms: Technical Advancement and Future Applications

PubMed Central

Kubota, Kengo

2013-01-01

Fluorescence in situ hybridization (FISH) has become a standard technique in environmental microbiology. More than 20 years have passed since this technique was first described, and it is currently used for the detection of ribosomal RNA, messenger RNA, and functional genes encoded on chromosomes. This review focuses on the advancement and applications of FISH combined with catalyzed reporter deposition (CARD, also known as tyramide signal amplification or TSA), in the detection of environmental microorganisms. Significant methodological improvements have been made in CARD-FISH technology, including its combination with other techniques and instruments. PMID:23124765

The application of fluorescence in situ hybridization in different ploidy levels cross-breeding of lily.

PubMed

Wang, Qing; Wang, Jingmao; Zhang, Yiying; Zhang, Yue; Xu, Shunchao; Lu, Yingmin

2015-01-01

21 crossing were conducted between Asiatic Lily with different ploidy levels, the results showed that the interploidy hybridization between diploid and tetraploid lilies was not as successful as intraploidy hybridization. Regardless of male sterility, triploid lilies could be used as female parents in the hybridization which the progenies were aneuploidy. 3x×4x crosses could be cultured more successfully than 3x×2x crosses. 45S rDNA was mapped on the chromosomes of seven Lilium species and their progenies using fluorescence in situ hybridization (FISH). FISH revealed six to sixteen 45S rDNA gene loci, and normally the sites were not in pairs. The asymmetry indexes of LA (Longiflorum hybrids × Asiatic hybrids) hybrids was higher than Asiatic hybrids, the evolution degree was LA hybrids > Asiatic hybrids. 45S rDNA distributed variably on chromosome 1-10 and 12 among Asiatic hybrids. Chromosome 1 had invariable sites of 45S rDNA in all Asiatic hybrids, which could be considered as the characteristic of Asiatic hybrids. LA hybrid 'Freya' had two sites of 45S rDNA on one homologous chromosome 5, and also it could be found in the progenies. The karyotype and fluorescence in situ hybridization with 45S rDNA as probe were applied to identify the different genotypes of 9 hybrids. Typical chromosomes with parental signal sites could be observed in all the genotypes of hybrids, it was confirmed that all the hybrids were true.

Mercury concentration in meconium and risk assessment of fish consumption among pregnant women in Taiwan.

PubMed

Jiang, Chuen-Bin; Yeh, Ching-Ying; Lee, Hung-Chang; Chen, Ming-Jun; Hung, Fang-Yu; Fang, Sheng-Shiung; Chien, Ling-Chu

2010-01-01

Meconium is a matrix that can be obtained easily and noninvasively and is useful for detecting antenatal fetal exposure to environmental toxins. Taiwan is an island with high fish consumption, and many pregnant women would like to enjoy the benefits of fish without jeopardizing their health or that of their child. The aim of this study is to assess the mercury concentration in meconium in relation to the health risk of mercury exposure. A total of 198 mother-infant pairs residing in the city of HsinChu were recruited for the study between January 2007 and June 2007. The average mean concentration of mercury in meconium was 79.2+/-7.3 ng g(-1) dry wt We use the Monte Carlo technique to assess the uncertainty in risk assessment and the impact of these uncertainties on the estimation of expected risk of mercury intake from fish in mothers. Based on the FAO/WHO's tolerable daily intake of methylmercury (0.23 microg kg(-1)d(-1)), we found that 17.3% and 14.0% of the daily mercury exposure estimated exceeded the reference dose for foreign-born and Taiwan-born mothers, respectively. We found that the mercury concentration in meconium was much higher than in other studies, except for one study done in Tagum in the Philippines where mercury is used in gold mining. This may be because Asia is the largest emitter of anthropogenic mercury, accounting for 53% of worldwide emissions. Sensitivity analysis suggests that mercury concentration in fish and the rate of ingesting fish may be the key parameters for governments offering risk management guidance to protect the health of mothers and unborn babies.

Assessment on dioxin-like compounds intake from various marine fish from Zhoushan Fishery, China.

PubMed

Wang, Xiangyong; Zhang, Hongxia; Zhang, Lei; Zhong, Kai; Shang, Xiaohong; Zhao, Yunfeng; Tong, Zhendong; Yu, Xinwei; Li, Jingguang; Wu, Yongning

2015-01-01

Sea fish consuming is an important intake source of dioxin-like compounds, especially for the coastal residents. To assess the intake levels of these contaminants from sea fish and to provide risk-based consumption advice, concentrations of 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and 12 dioxin-like polychlorinated biphenyls (dl-PCBs) were measured in 32 commonly consumed fish species from Zhoushan Fishery, China. Due to the different accumulation influenced by fat content, feed habits and living zone in the sea area, the levels of PCDDs, PCDFs and dl-PCBs in different fish species varied significantly ranging from 0.002 to 0.078pg WHO-TEQ/g fresh weight, from 0.002 to 0.553pg WHO-TEQ/g fresh weight and from 0.003 to 2.059pg WHO-TEQ/g fresh weight, respectively. Based on mean fish consuming rate in China, the estimated maximum possible dioxin-like compounds intake through different fish species ranged from 0.26 to 65.61pgTEQkg(-1)bwmonth(-1). Bullet mackerel has the highest monthly intake level which was much higher than other fish species and very close to the provisional tolerable monthly intake (70pgTEQkg(-)(1)bwmonth(-)(1)) proposed by the Joint FAO/WHO Expert Committee on Food Additives. Hence, comparing to other fish species, the consumption of Bullet mackerel from Zhoushan Fishery should be cautious to reduce the potential health risk. Copyright © 2014 Elsevier Ltd. All rights reserved.

Indices for assessing coral reef fish biodiversity: the need for a change in habits.

PubMed

Loiseau, Nicolas; Gaertner, Jean-Claude

2015-09-01

We present the first representative and quantified overview of the indices used worldwide for assessing the biodiversity of coral reef fishes. On this basis, we discuss the suitability and drawbacks of the indices most widely used in the assessment of coral fish biodiversity. An extensive and systematic survey of the literature focused on coral reef fish biodiversity was conducted from 1990 up to the present. We found that the multicomponent aspect of biodiversity, which is considered as a key feature of biodiversity for numerous terrestrial and marine ecosystems, has been poorly taken into account in coral reef fish studies. Species richness is still strongly dominant while other diversity components, such as functional diversity, are underestimated even when functional information is available. We also demonstrate that the reason for choosing particular indices is often unclear, mainly based on empirical rationales and/or the reproduction of widespread habits, but generally with no clear relevance with regard to the aims of the studies. As a result, the most widely used indices (species richness, Shannon, etc.) would appear to be poorly suited to meeting the main challenges facing the monitoring of coral reef fish biodiversity in the future. Our results clearly show that coral reef scientists should rather take advantage of the multicomponent aspect of biodiversity. To facilitate this approach, we propose general guidelines to serve as a basis for the selection of indices that provide complementary and relevant information for monitoring the response of coral reef fish biodiversity in the face of structuring factors (natural or anthropic). The aim of these guidelines was to achieve a better match between the properties of the selected indices and the context of each study (e.g. expected effect of the main structuring factors, nature of data available).

Assessing Historical Fish Community Composition Using Surveys, Historical Collection Data, and Species Distribution Models

PubMed Central

Labay, Ben; Cohen, Adam E.; Sissel, Blake; Hendrickson, Dean A.; Martin, F. Douglas; Sarkar, Sahotra

2011-01-01

Accurate establishment of baseline conditions is critical to successful management and habitat restoration. We demonstrate the ability to robustly estimate historical fish community composition and assess the current status of the urbanized Barton Creek watershed in central Texas, U.S.A. Fish species were surveyed in 2008 and the resulting data compared to three sources of fish occurrence information: (i) historical records from a museum specimen database and literature searches; (ii) a nearly identical survey conducted 15 years earlier; and (iii) a modeled historical community constructed with species distribution models (SDMs). This holistic approach, and especially the application of SDMs, allowed us to discover that the fish community in Barton Creek was more diverse than the historical data and survey methods alone indicated. Sixteen native species with high modeled probability of occurrence within the watershed were not found in the 2008 survey, seven of these were not found in either survey or in any of the historical collection records. Our approach allowed us to more rigorously establish the true baseline for the pre-development fish fauna and then to more accurately assess trends and develop hypotheses regarding factors driving current fish community composition to better inform management decisions and future restoration efforts. Smaller, urbanized freshwater systems, like Barton Creek, typically have a relatively poor historical biodiversity inventory coupled with long histories of alteration, and thus there is a propensity for land managers and researchers to apply inaccurate baseline standards. Our methods provide a way around that limitation by using SDMs derived from larger and richer biodiversity databases of a broader geographic scope. Broadly applied, we propose that this technique has potential to overcome limitations of popular bioassessment metrics (e.g., IBI) to become a versatile and robust management tool for determining status of

Assessing historical fish community composition using surveys, historical collection data, and species distribution models.

PubMed

Labay, Ben; Cohen, Adam E; Sissel, Blake; Hendrickson, Dean A; Martin, F Douglas; Sarkar, Sahotra

2011-01-01

Accurate establishment of baseline conditions is critical to successful management and habitat restoration. We demonstrate the ability to robustly estimate historical fish community composition and assess the current status of the urbanized Barton Creek watershed in central Texas, U.S.A. Fish species were surveyed in 2008 and the resulting data compared to three sources of fish occurrence information: (i) historical records from a museum specimen database and literature searches; (ii) a nearly identical survey conducted 15 years earlier; and (iii) a modeled historical community constructed with species distribution models (SDMs). This holistic approach, and especially the application of SDMs, allowed us to discover that the fish community in Barton Creek was more diverse than the historical data and survey methods alone indicated. Sixteen native species with high modeled probability of occurrence within the watershed were not found in the 2008 survey, seven of these were not found in either survey or in any of the historical collection records. Our approach allowed us to more rigorously establish the true baseline for the pre-development fish fauna and then to more accurately assess trends and develop hypotheses regarding factors driving current fish community composition to better inform management decisions and future restoration efforts. Smaller, urbanized freshwater systems, like Barton Creek, typically have a relatively poor historical biodiversity inventory coupled with long histories of alteration, and thus there is a propensity for land managers and researchers to apply inaccurate baseline standards. Our methods provide a way around that limitation by using SDMs derived from larger and richer biodiversity databases of a broader geographic scope. Broadly applied, we propose that this technique has potential to overcome limitations of popular bioassessment metrics (e.g., IBI) to become a versatile and robust management tool for determining status of

Assessment of tolerant sunfish populations (Lepomis sp.) inhabiting selenium-laden coal ash effluents. 3. Serum chemistry and fish health indicators.

PubMed

Lohner, T W; Reash, R J; Willet, V E; Fletcher, J

2001-11-01

Sunfish were collected from fly ash discharge-receiving streams to assess the possible effects of exposure to elevated selenium. Concentrations of selenium, copper, and arsenic were statistically higher in fish tissue (liver) samples from effluent-exposed fish than in reference fish. Several biomarkers were indicative of metal exposure and effect. Plasma protein levels and cholesterol levels were significantly lower in exposed fish, indicating nutritional stress. Ion levels (i.e., K) increased with exposure to ash pond metals, indicating possible gill damage. Fish from the receiving streams also had increased serum glucose and osmolality indicating possible acute stress due to sampling. Fish health assessments revealed a lower incidence of fin erosion, kidney discoloration, urolithiasis or nephrocalcinosis, liver discoloration, and parasites in exposed fish and a higher incidence of skin, eye, and gill aberrations. Condition factors of exposed fish were correlated with biomarker response and were the same as or lower than those of reference fish, but not related to selenium levels. Although several serum biochemical indicators differed between the ash pond-receiving stream and reference sites, pollutant exposure was apparently not sufficient to cause functional damage to critical organ systems.

Dispersal and selection mediate hybridization between a native and invasive species

USGS Publications Warehouse

Kovach, Ryan P.; Muhlfeld, Clint C.; Boyer, Matthew C.; Lowe, Winsor H.; Allendorf, Fred W.; Luikart, Gordon

2015-01-01

Hybridization between native and non-native species has serious biological consequences, but our understanding of how dispersal and selection interact to influence invasive hybridization is limited. Here, we document the spread of genetic introgression between a native (Oncorhynchus clarkii) and invasive (Oncorhynchus mykiss) trout, and identify the mechanisms influencing genetic admixture. In two populations inhabiting contrasting environments, non-native admixture increased rapidly from 1984 to 2007 and was driven by surprisingly consistent processes. Individual admixture was related to two phenotypic traits associated with fitness: size at spawning and age of juvenile emigration. Fish with higher non-native admixture were larger and tended to emigrate at a younger age—relationships that are expected to confer fitness advantages to hybrid individuals. However, strong selection against non-native admixture was evident across streams and cohorts (mean selection coefficient against genotypes with non-native alleles (s) ¼ 0.60; s.e. ¼ 0.10). Nevertheless, hybridization was promoted in both streams by the continuous immigration of individuals with high levels of non-native admixture from other hybrid source populations. Thus, antagonistic relationships between dispersal and selection are mediating invasive hybridization between these fish, emphasizing that data on dispersal and natural selection are needed to fully understand the dynamics of introgression between native and non-native species. .

GENETIC DIVERSITY OF STREAM FISH IN THE MID-ATLANTIC INTEGRATED ASSESSMENT AREA

EPA Science Inventory

In this report, we describe the results of research to assess the genetics of three stream fish species in the Mid-Atlantic region. This research had three specific goals. First, we sought to examine the utility of genetic analysis for purposes of taxonomic identification. Bioa...

High-resolution tyramide-FISH mapping of markers tightly linked to the male-fertility restoration (Ms) locus of onion

USDA-ARS?s Scientific Manuscript database

Fluorescence in situ hybridization (FISH) has not been readily exploited for physical mapping of molecular markers in plants due to the technical challenge to visualize small single-copy probes. Signal amplification using tyramide (tyr) FISH can increase sensitivity up to 100 fold. We used tyr-FISH ...

Arsenic Speciation in Fish Products and Seafood as a Prerequisite for Proper Risk Assessment

PubMed Central

Orletti, Roberta; Chessa, Giannina; Carloni, Cristiano; Griffoni, Francesco; Palombo, Paolo; Velieri, Francesco

2015-01-01

The Boi Cerbus lagoon, facing a mining and industrial site in Sardinia (Italy), is an important fishing area for the local population. Previous studies showed high concentrations of total arsenic (Astot) in fish, molluscs and crustaceans sampled in the lagoon, and a possible exceeding of the provisional tolerable weekly intake set by the Joint FAO/WHO Expert Committee on Food Additives by some local consumer groups. However, the percentage of inorganic As (Asinorg) should be known for a correct assessment of potential risk, as its toxicity is much higher than that of the organic forms. Eighty samples of 14 different species of fish, molluscs and crustaceans, sampled in the Boi Cerbus lagoon in 3 different seasons (winter, spring and summer), were analysed for Astot by inductively coupled plasma mass spectrometry (ICP-MS) and Asinorg by high performance liquid chromatography-ICP-MS. All the data obtained from the analysis were statistically processed to evaluate significant differences based on season, taxon and habitat, in preparation for a subsequent risk assessment. PMID:27800381

Hybridization-based detection of Helicobacter pylori at human body temperature using advanced locked nucleic acid (LNA) probes.

PubMed

Fontenete, Sílvia; Guimarães, Nuno; Leite, Marina; Figueiredo, Céu; Wengel, Jesper; Filipe Azevedo, Nuno

2013-01-01

The understanding of the human microbiome and its influence upon human life has long been a subject of study. Hence, methods that allow the direct detection and visualization of microorganisms and microbial consortia (e.g. biofilms) within the human body would be invaluable. In here, we assessed the possibility of developing a variant of fluorescence in situ hybridization (FISH), named fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) and 2'-O-methyl RNAs (2'OMe) with two types of backbone linkages (phosphate or phosphorothioate), we were able to successfully identify two probes that hybridize at 37 °C with high specificity and sensitivity for H. pylori, both in pure cultures and in gastric biopsies. Furthermore, the use of this type of probes implied that toxic compounds typically used in FISH were either found to be unnecessary or could be replaced by a non-toxic substitute. We show here for the first time that the use of advanced LNA probes in FIVH conditions provides an accurate, simple and fast method for H. pylori detection and location, which could be used in the future for potential in vivo applications either for this microorganism or for others.

Hybridization-Based Detection of Helicobacter pylori at Human Body Temperature Using Advanced Locked Nucleic Acid (LNA) Probes

PubMed Central

Fontenete, Sílvia; Guimarães, Nuno; Leite, Marina; Figueiredo, Céu; Wengel, Jesper; Filipe Azevedo, Nuno

2013-01-01

The understanding of the human microbiome and its influence upon human life has long been a subject of study. Hence, methods that allow the direct detection and visualization of microorganisms and microbial consortia (e.g. biofilms) within the human body would be invaluable. In here, we assessed the possibility of developing a variant of fluorescence in situ hybridization (FISH), named fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) and 2’-O-methyl RNAs (2’OMe) with two types of backbone linkages (phosphate or phosphorothioate), we were able to successfully identify two probes that hybridize at 37 °C with high specificity and sensitivity for H. pylori, both in pure cultures and in gastric biopsies. Furthermore, the use of this type of probes implied that toxic compounds typically used in FISH were either found to be unnecessary or could be replaced by a non-toxic substitute. We show here for the first time that the use of advanced LNA probes in FIVH conditions provides an accurate, simple and fast method for H. pylori detection and location, which could be used in the future for potential in vivo applications either for this microorganism or for others. PMID:24278398

Linking hydrologic, physical and chemical habitat environments for the potential assessment of fish community rehabilitation in a developing city

NASA Astrophysics Data System (ADS)

Zhao, C. S.; Yang, S. T.; Liu, C. M.; Dou, T. W.; Yang, Z. L.; Yang, Z. Y.; Liu, X. L.; Xiang, H.; Nie, S. Y.; Zhang, J. L.; Mitrovic, S. M.; Yu, Q.; Lim, R. P.

2015-04-01

Aquatic ecological rehabilitation is increasingly attracting considerable public and research attention. An effective method that requires less data and expertise would help in the assessment of rehabilitation potential and in the monitoring of rehabilitation activities as complicated theories and excessive data requirements on assemblage information make many current assessment models expensive and limit their wide use. This paper presents an assessment model for restoration potential which successfully links hydrologic, physical and chemical habitat factors to fish assemblage attributes drawn from monitoring datasets on hydrology, water quality and fish assemblages at a total of 144 sites, where 5084 fish were sampled and tested. In this model three newly developed sub-models, integrated habitat index (IHSI), integrated ecological niche breadth (INB) and integrated ecological niche overlap (INO), are established to study spatial heterogeneity of the restoration potential of fish assemblages based on gradient methods of habitat suitability index and ecological niche models. To reduce uncertainties in the model, as many fish species as possible, including important native fish, were selected as dominant species with monitoring occurring over several seasons to comprehensively select key habitat factors. Furthermore, a detrended correspondence analysis (DCA) was employed prior to a canonical correspondence analysis (CCA) of the data to avoid the "arc effect" in the selection of key habitat factors. Application of the model to data collected at Jinan City, China proved effective reveals that three lower potential regions that should be targeted in future aquatic ecosystem rehabilitation programs. They were well validated by the distribution of two habitat parameters: river width and transparency. River width positively influenced and transparency negatively influenced fish assemblages. The model can be applied for monitoring the effects of fish assemblage restoration

Assessing fishing and marine biodiversity changes using fishers' perceptions: the Spanish Mediterranean and Gulf of Cadiz case study.

PubMed

Coll, Marta; Carreras, Marta; Ciércoles, Cristina; Cornax, Maria-José; Gorelli, Giulia; Morote, Elvira; Sáez, Raquel

2014-01-01

The expansion of fishing activities has intensively transformed marine ecosystems worldwide. However, available time series do not frequently cover historical periods. Fishers' perceptions were used to complement data and characterise changes in fishing activity and exploited ecosystems in the Spanish Mediterranean Sea and Gulf of Cadiz. Fishers' interviews were conducted in 27 fishing harbours of the area, and included 64 fishers from ages between 20 to >70 years old to capture the experiences and memories of various generations. Results are discussed in comparison with available independent information using stock assessments and international convention lists. According to fishers, fishing activity substantially evolved in the area with time, expanding towards deeper grounds and towards areas more distant from the coast. The maximum amount of catch ever caught and the weight of the largest species ever captured inversely declined with time. Fishers (70%) cited specific fishing grounds where depletion occurred. They documented ecological changes of marine biodiversity during the last half of the century: 94% reported the decline of commercially important fish and invertebrates and 61% listed species that could have been extirpated, with frequent mentions to cartilaginous fish. Declines and extirpations were in line with available quantitative evaluations from stock assessments and international conventions, and were likely linked to fishing impacts. Conversely, half of interviewed fishers claimed that several species had proliferated, such as cephalopods, jellyfish, and small-sized fish. These changes were likely related to trophic cascades due to fishing and due to climate change effects. The species composition of depletions, local extinctions and proliferations showed differences by region suggesting that regional dynamics are important when analysing biodiversity changes. Using fishers' perceptions, fishing and ecological changes in the study area were

Assessing Fishing and Marine Biodiversity Changes Using Fishers' Perceptions: The Spanish Mediterranean and Gulf of Cadiz Case Study

PubMed Central

Coll, Marta; Carreras, Marta; Ciércoles, Cristina; Cornax, Maria-José; Gorelli, Giulia; Morote, Elvira; Sáez, Raquel

2014-01-01

Background The expansion of fishing activities has intensively transformed marine ecosystems worldwide. However, available time series do not frequently cover historical periods. Methodology Fishers' perceptions were used to complement data and characterise changes in fishing activity and exploited ecosystems in the Spanish Mediterranean Sea and Gulf of Cadiz. Fishers' interviews were conducted in 27 fishing harbours of the area, and included 64 fishers from ages between 20 to >70 years old to capture the experiences and memories of various generations. Results are discussed in comparison with available independent information using stock assessments and international convention lists. Principal Findings According to fishers, fishing activity substantially evolved in the area with time, expanding towards deeper grounds and towards areas more distant from the coast. The maximum amount of catch ever caught and the weight of the largest species ever captured inversely declined with time. Fishers (70%) cited specific fishing grounds where depletion occurred. They documented ecological changes of marine biodiversity during the last half of the century: 94% reported the decline of commercially important fish and invertebrates and 61% listed species that could have been extirpated, with frequent mentions to cartilaginous fish. Declines and extirpations were in line with available quantitative evaluations from stock assessments and international conventions, and were likely linked to fishing impacts. Conversely, half of interviewed fishers claimed that several species had proliferated, such as cephalopods, jellyfish, and small-sized fish. These changes were likely related to trophic cascades due to fishing and due to climate change effects. The species composition of depletions, local extinctions and proliferations showed differences by region suggesting that regional dynamics are important when analysing biodiversity changes. Conclusions/Significance Using fishers

A fish-based index of biotic integrity to assess intermittent headwater streams in Wisconsin, USA.

PubMed

Lyons, John

2006-11-01

I developed a fish-based index of biotic integrity (IBI) to assess environmental quality in intermittent headwater streams in Wisconsin, USA. Backpack electrofishing and habitat surveys were conducted four times on 102 small (watershed area 1.7-41.5 km(2)), cool or warmwater (maximum daily mean water temperature > or = 22 C), headwater streams in spring and late summer/fall 2000 and 2001. Despite seasonal and annual changes in stream flow and habitat volume, there were few significant temporal trends in fish attributes. Analysis of 36 least-impacted streams indicated that fish were too scarce to calculate an IBI at stations with watershed areas less than 4 km(2) or at stations with watershed areas from 4-10 km(2) if stream gradient exceeded 10 m/km (1% slope). For streams with sufficient fish, potential fish attributes (metrics) were not related to watershed size or gradient. Seven metrics distinguished among streams with low, agricultural, and urban human impacts: numbers of native, minnow (Cyprinidae), headwater-specialist, and intolerant (to environmental degradation) species; catches of all fish excluding species tolerant of environmental degradation and of brook stickleback (Culaea inconstans) per 100 m stream length; and percentage of total individuals with deformities, eroded fins, lesions, or tumors. These metrics were used in the final IBI, which ranged from 0 (worst) to 100 (best). The IBI accurately assessed the environmental quality of 16 randomly chosen streams not used in index development. Temporal variation in IBI scores in the absence of changes in environmental quality was not related to season, year, or type of human impact and was similar in magnitude to variation reported for other IBI's.

Preliminary assessment of factors influencing riverine fish communities in Massachusetts.

USGS Publications Warehouse

Armstrong, David S.; Richards, Todd A.; Brandt, Sara L.

2010-01-01

The U.S. Geological Survey, in cooperation with the Massachusetts Department of Conservation and Recreation (MDCR), Massachusetts Department of Environmental Protection (MDEP), and the Massachusetts Department of Fish and Game (MDFG), conducted a preliminary investigation of fish communities in small- to medium-sized Massachusetts streams. The objective of this investigation was to determine relations between fish-community characteristics and anthropogenic alteration, including flow alteration and impervious cover, relative to the effect of physical basin and land-cover (environmental) characteristics. Fish data were obtained for 756 fish-sampling sites from the Massachusetts Division of Fisheries and Wildlife fish-community database. A review of the literature was used to select a set of fish metrics responsive to flow alteration. Fish metrics tested include two fish-community metrics (fluvial-fish relative abundance and fluvial-fish species richness), and five indicator species metrics (relative abundance of brook trout, blacknose dace, fallfish, white sucker, and redfin pickerel). Streamflows were simulated for each fish-sampling site using the Sustainable Yield Estimator application (SYE). Daily streamflows and the SYE water-use database were used to determine a set of indicators of flow alteration, including percent alteration of August median flow, water-use intensity, and withdrawal and return-flow fraction. The contributing areas to the fish-sampling sites were delineated and used with a Geographic Information System (GIS) to determine a set of environmental characteristics, including elevation, basin slope, percent sand and gravel, percent wetland, and percent open water, and a set of anthropogenic-alteration variables, including impervious cover and dam density. Two analytical techniques, quantile regression and generalized linear modeling, were applied to determine the association between fish-response variables and the selected environmental and

Real-time assessment of hybridization between wolves and dogs: combining noninvasive samples with ancestry informative markers.

PubMed

Godinho, Raquel; López-Bao, José Vicente; Castro, Diana; Llaneza, Luís; Lopes, Susana; Silva, Pedro; Ferrand, Nuno

2015-03-01

Wolves and dogs provide a paradigmatic example of the ecological and conservation implications of hybridization events between wild and domesticated forms. However, our understanding of such implications has been traditionally hampered by both high genetic similarity and the difficulties in obtaining tissue samples (TS), which limit our ability to assess ongoing hybridization events. To assess the occurrence and extension of hybridization in a pack of wolf-dog hybrids in northwestern Iberia, we compared the power of 52 nuclear markers implemented on TS with a subset of 13 ancestry informative markers (AIMs) typed in noninvasive samples (NIS). We demonstrate that the 13 AIMs are as accurate as the 52 markers that were chosen without regard to the power to differentiate between wolves and dogs, also having the advantage of being rapidly screened on NIS. The efficiency of AIMs significantly outperformed ten random sets of similar size and an additional commercial set of 18 markers. Bayesian clustering analysis implemented on AIMs and NIS identified nine hybrids, two wolves and two dogs. Four hybrids were unambiguously assigned to F1xWolf backcrosses. Our approach (AIMs + NIS) overcomes previous difficulties related to sample availability and informative power of markers, allowing a quick identification of wolf-dog hybrids in the first phases of hybridization episodes. This provides managers with a reliable tool to evaluate hybridization and estimate the success of their actions. This approach may be easily adapted for other pairs of wild/domesticated species, thus improving our understanding of the introgression of domestication genes into natural populations. © 2014 John Wiley & Sons Ltd.

Length-Based Assessment of Coral Reef Fish Populations in the Main and Northwestern Hawaiian Islands

PubMed Central

Nadon, Marc O.; Ault, Jerald S.; Williams, Ivor D.; Smith, Steven G.; DiNardo, Gerard T.

2015-01-01

The coral reef fish community of Hawaii is composed of hundreds of species, supports a multimillion dollar fishing and tourism industry, and is of great cultural importance to the local population. However, a major stock assessment of Hawaiian coral reef fish populations has not yet been conducted. Here we used the robust indicator variable “average length in the exploited phase of the population (L¯)”, estimated from size composition data from commercial fisheries trip reports and fishery-independent diver surveys, to evaluate exploitation rates for 19 Hawaiian reef fishes. By and large, the average lengths obtained from diver surveys agreed well with those from commercial data. We used the estimated exploitation rates coupled with life history parameters synthesized from the literature to parameterize a numerical population model and generate stock sustainability metrics such as spawning potential ratios (SPR). We found good agreement between predicted average lengths in an unfished population (from our population model) and those observed from diver surveys in the largely unexploited Northwestern Hawaiian Islands. Of 19 exploited reef fish species assessed in the main Hawaiian Islands, 9 had SPRs close to or below the 30% overfishing threshold. In general, longer-lived species such as surgeonfishes, the redlip parrotfish (Scarus rubroviolaceus), and the gray snapper (Aprion virescens) had the lowest SPRs, while short-lived species such as goatfishes and jacks, as well as two invasive species (Lutjanus kasmira and Cephalopholis argus), had SPRs above the 30% threshold. PMID:26267473

The Application of Fluorescence In Situ Hybridization in Different Ploidy Levels Cross-Breeding of Lily

PubMed Central

Wang, Qing; Wang, Jingmao; Zhang, Yiying; Zhang, Yue; Xu, Shunchao; Lu, Yingmin

2015-01-01

21 crossing were conducted between Asiatic Lily with different ploidy levels, the results showed that the interploidy hybridization between diploid and tetraploid lilies was not as successful as intraploidy hybridization. Regardless of male sterility, triploid lilies could be used as female parents in the hybridization which the progenies were aneuploidy. 3x×4x crosses could be cultured more successfully than 3x×2x crosses. 45S rDNA was mapped on the chromosomes of seven Lilium species and their progenies using fluorescence in situ hybridization (FISH). FISH revealed six to sixteen 45S rDNA gene loci, and normally the sites were not in pairs. The asymmetry indexes of LA (Longiflorum hybrids × Asiatic hybrids) hybrids was higher than Asiatic hybrids, the evolution degree was LA hybrids > Asiatic hybrids. 45S rDNA distributed variably on chromosome 1-10 and 12 among Asiatic hybrids. Chromosome 1 had invariable sites of 45S rDNA in all Asiatic hybrids, which could be considered as the characteristic of Asiatic hybrids. LA hybrid ‘Freya’ had two sites of 45S rDNA on one homologous chromosome 5, and also it could be found in the progenies. The karyotype and fluorescence in situ hybridization with 45S rDNA as probe were applied to identify the different genotypes of 9 hybrids. Typical chromosomes with parental signal sites could be observed in all the genotypes of hybrids, it was confirmed that all the hybrids were true. PMID:26010356

Assessing speech perception in children with cochlear implants using a modified hybrid visual habituation procedure.

PubMed

Core, Cynthia; Brown, Janean W; Larsen, Michael D; Mahshie, James

2014-01-01

The objectives of this research were to determine whether an adapted version of a Hybrid Visual Habituation procedure could be used to assess speech perception of phonetic and prosodic features of speech (vowel height, lexical stress, and intonation) in individual pre-school-age children who use cochlear implants. Nine children ranging in age from 3;4 to 5;5 participated in this study. Children were prelingually deaf and used cochlear implants and had no other known disabilities. Children received two speech feature tests using an adaptation of a Hybrid Visual Habituation procedure. Seven of the nine children demonstrated perception of at least one speech feature using this procedure using results from a Bayesian linear regression analysis. At least one child demonstrated perception of each speech feature using this assessment procedure. An adapted version of the Hybrid Visual Habituation Procedure with an appropriate statistical analysis provides a way to assess phonetic and prosodicaspects of speech in pre-school-age children who use cochlear implants.

Using Tournament Angler Data to Rapidly Assess the Invasion Status of Alien Sport Fishes (Micropterus spp.) in Southern Africa.

PubMed

Hargrove, John S; Weyl, Olaf L F; Allen, Micheal S; Deacon, Neil R

2015-01-01

Fishes are one of the most commonly introduced aquatic taxa worldwide, and invasive fish species pose threats to biodiversity and ecosystem function in recipient waters. Considerable research efforts have focused on predicting the invasibility of different fish taxa; however, accurate records detailing the establishment and spread of invasive fishes are lacking for large numbers of fish around the globe. In response to these data limitations, a low-cost method of cataloging and quantifying the temporal and spatial status of fish invasions was explored. Specifically, angler catch data derived from competitive bass angling tournaments was used to document the distribution of 66 non-native populations of black bass (Micropterus spp.) in southern Africa. Additionally, catch data from standardized tournament events were used to assess the abundance and growth of non-native bass populations in southern Africa relative to their native distribution (southern and eastern United States). Differences in metrics of catch per unit effort (average number of fish retained per angler per day), daily bag weights (the average weight of fish retained per angler), and average fish weight were assessed using catch data from 14,890 angler days of tournament fishing (11,045 days from South Africa and Zimbabwe; 3,845 days from the United States). No significant differences were found between catch rates, average daily bag weight, or the average fish weight between countries, suggesting that bass populations in southern Africa reach comparable sizes and numbers relative to waters in their native distribution. Given the minimal cost associated with data collection (i.e. records are collected by tournament organizers), the standardized nature of the events, and consistent bias (i.e. selection for the biggest fish in a population), the use of angler catch data represents a novel approach to infer the status and distribution of invasive sport fish.

Using Tournament Angler Data to Rapidly Assess the Invasion Status of Alien Sport Fishes (Micropterus spp.) in Southern Africa

PubMed Central

Hargrove, John S.; Weyl, Olaf L. F.; Allen, Micheal S.; Deacon, Neil R.

2015-01-01

Fishes are one of the most commonly introduced aquatic taxa worldwide, and invasive fish species pose threats to biodiversity and ecosystem function in recipient waters. Considerable research efforts have focused on predicting the invasibility of different fish taxa; however, accurate records detailing the establishment and spread of invasive fishes are lacking for large numbers of fish around the globe. In response to these data limitations, a low-cost method of cataloging and quantifying the temporal and spatial status of fish invasions was explored. Specifically, angler catch data derived from competitive bass angling tournaments was used to document the distribution of 66 non-native populations of black bass (Micropterus spp.) in southern Africa. Additionally, catch data from standardized tournament events were used to assess the abundance and growth of non-native bass populations in southern Africa relative to their native distribution (southern and eastern United States). Differences in metrics of catch per unit effort (average number of fish retained per angler per day), daily bag weights (the average weight of fish retained per angler), and average fish weight were assessed using catch data from 14,890 angler days of tournament fishing (11,045 days from South Africa and Zimbabwe; 3,845 days from the United States). No significant differences were found between catch rates, average daily bag weight, or the average fish weight between countries, suggesting that bass populations in southern Africa reach comparable sizes and numbers relative to waters in their native distribution. Given the minimal cost associated with data collection (i.e. records are collected by tournament organizers), the standardized nature of the events, and consistent bias (i.e. selection for the biggest fish in a population), the use of angler catch data represents a novel approach to infer the status and distribution of invasive sport fish. PMID:26047487

Do scientists and fishermen collect the same size fish? Possible implications for exposure assessment.

PubMed

Burger, Joanna; Gochfeld, Michael; Burke, Sean; Jeitner, Christian W; Jewett, Stephen; Snigaroff, Daniel; Snigaroff, Ronald; Stamm, Tim; Harper, Shawn; Hoberg, Max; Chenelot, Heloise; Patrick, Robert; Volz, Conrad D; Weston, James

2006-05-01

research/commercial trawler operated under charter to the National Oceanographic and Atmospheric Administration (NOAA). The fish selected for sampling, including those caught commercially in the Bering Sea, represented different trophic levels, and are species regularly caught by Aleuts while fishing near their villages. Not all fish were caught by all three groups. There were no significant differences in length and weight for five species of fish caught by Aleuts, scientists, and fisheries trawls, and for an additional 3 species caught only by the Aleut and scientist teams. There were small, but significant, differences in the sizes of rock greenling (Hexagrammos lagocephalus) and red Irish lord (Hemilepidotus hemilepidotus) caught by the scientist and Aleut fishermen. No scientists caught rock greenling using poles; those speared by the divers were significantly smaller than those caught by the Aleuts. Further, there were no differences in the percent of males in the samples as a function of fishing method or type of fishermen, except for rockfish and red Irish lord. These data suggest that if scientists collect fish in the same manner as subsistence fishermen (in this case, using fishing rods from boats), they can collect the same-sized fish. The implications for exposure and risk assessment are that scientists should either engage subsistence and recreational fishermen to collect fish for analysis, or mimic their fishing methods to ensure that the fish collected are similar in size and weight to those being caught and consumed by these groups. Further, total length, standard length, and weight were highly correlated for all species of fish, suggesting that risk assessors could rely on recreational and commercial fishermen to measure total lengths for the purpose of correlating mercury levels with known size/mercury level relationships. Our data generally demonstrate that the scientists and trawlers can collect the same size fish as those caught by Aleuts, making

Risk and toxicity assessments of heavy metals in sediments and fishes from the Yangtze River and Taihu Lake, China.

PubMed

Fu, Jie; Hu, Xin; Tao, Xiancong; Yu, Hongxia; Zhang, Xiaowei

2013-11-01

Heavy metal pollution is one of the most serous environmental issues globally. To evaluate the metal pollution in Jiangsu Province of China, the total concentrations of heavy metals in sediments and fishes from the Yangtze River and Taihu Lake were analyzed. Ecological risk of sediments and human health risk of fish consumption were assessed respectively. Furthermore, toxicity of samples on expression of the stress responsive genes was evaluated using microbial live cell-array method. The results showed that the heavy metals concentrations in sediments from the Yangtze River were much higher than those in sediments from the Taihu Lake. However, the fishes from the Taihu Lake had higher concentrations of heavy metals than fishes from the Yangtze River. Ecological risk evaluation showed that the heavy metal contaminants in sediments from the Yangtze River posed higher risk of adverse ecological effects, while sediments from the study areas of Taihu Lake were relatively safe. Health risk assessment suggested that the heavy metals in fishes of both Yangtze River and Taihu Lake might have risk of adverse health effects to human. The toxicity assessment indicated that the heavy metals in these sediments and fishes showed transcriptional effects on the selected 21 stress responsive genes, which were involved in the pathways of DNA damage response, chemical stress, and perturbations of electron transport. Together, this field investigation combined with chemical analysis, risk assessment and toxicity bioassay would provide useful information on the heavy metal pollution in Jiangsu Province. Copyright © 2013 Elsevier Ltd. All rights reserved.

Total and inorganic arsenic in fish, seafood and seaweeds--exposure assessment.

PubMed

Mania, Monika; Rebeniak, Małgorzata; Szynal, Tomasz; Wojciechowska-Mazurek, Maria; Starska, Krystyna; Ledzion, Ewa; Postupolski, Jacek

2015-01-01

According to the European Food Safety Authority (EFSA), fish, seafood and seaweeds are foodstuffs that significantly contribute to dietary arsenic intake. With the exception of some algal species, the dominant compounds of arsenic in such food products are the less toxic organic forms. Both the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and EFSA recommend that speciation studies be performed to determine the different chemical forms in which arsenic is present in food due to the differences in their toxicity. Knowing such compositions can thus enable a complete exposure assessment to be made. Determination of total and inorganic arsenic contents in fish, their products, seafood and seaweeds present on the Polish market. This was then followed by an exposure assessment of consumers to inorganic arsenic in these foodstuffs. Total and inorganic arsenic was determined in 55 samples of fish, their products, seafood as well as seaweeds available on the market. The analytical method was hydride generation atomic absorption spectrometry (HGAAS), after dry ashing of samples and reduction of arsenic to arsenic hydride using sodium borohydride. In order to isolate only the inorganic forms of arsenic prior to mineralisation, samples were subjected to concentrated HCl hydrolysis, followed by reduction with hydrobromic acid and hydrazine sulphate after which triple chloroform extractions and triple 1M HCl re-extractions were performed. Exposure of adults was estimated in relation to the Benchmark Dose Lower Confidence Limit (BMDL0.5) as set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) that resulted in a 0.5% increase in lung cancer (3.0 μg/kg body weight (b.w.) per day). Mean total arsenic content from all investigated fish samples was 0.46 mg/kg (90th percentile 0.94 mg/kg), whilst the inorganic arsenic content never exceeded the detection limit of the analytical method used (0.025 mg/kg). In fish products, mean total arsenic concentration was

DEVELOPMENT OF A FISH INDEX OF BIOTIC INTEGRITY FOR ASSESSING WADEABLE STREAMS IN WEST VIRGINIA

EPA Science Inventory

We used Environmental Monitoring Assessment Program (EMAP) data from the Mid-Atlantic Highlands Assessment (MAHA) to develop an Index of Biotic Integrity (IBI) for West Virginia (WV). Standard techniques were used to evaluate a candidate pool of 58 fish assemblage metrics. Resp...

Monitoring fish distributions along electrofishing segments

USGS Publications Warehouse

Miranda, Leandro E.

2014-01-01

Electrofishing is widely used to monitor fish species composition and relative abundance in streams and lakes. According to standard protocols, multiple segments are selected in a body of water to monitor population relative abundance as the ratio of total catch to total sampling effort. The standard protocol provides an assessment of fish distribution at a macrohabitat scale among segments, but not within segments. An ancillary protocol was developed for assessing fish distribution at a finer scale within electrofishing segments. The ancillary protocol was used to estimate spacing, dispersion, and association of two species along shore segments in two local reservoirs. The added information provided by the ancillary protocol may be useful for assessing fish distribution relative to fish of the same species, to fish of different species, and to environmental or habitat characteristics.

Assessing Changes in Amphibian Population Dynamics Following Experimental Manipulations of Introduced Fish

Treesearch

Karen L. Pope

2008-01-01

Sport-fish introductions are now recognized as an important cause of amphibian decline, but few researchers have quantified the demographic responses of amphibians to current options in fisheries management designed to minimize effects on sensitive amphibians. Demographic analyses with mark–recapture data allow researchers to assess the relative importance of...

Multi-Ecosystem Assessment of Mercury Bioaccumulation in Fishes: Habitat, Landscape, and Biogeochemical Drivers of Fish Mercury

NASA Astrophysics Data System (ADS)

Eagles-Smith, C.; Ackerman, J.; Herring, G.; Willacker, J.; Flanagan, C.

2014-12-01

Mercury (Hg) is a globally distributed contaminant that threatens ecosystem health across aquatic environments. The complexity of the Hg cycle and its primary drivers, coupled with dynamic food web processes that govern biomagnification, result in marked spatial variability in Hg bioaccumulation across aquatic ecosystems. However, it is unclear if patterns of bioaccumulation are consistent in magnitude and direction across ecosystem types. We synthesized data from several studies spanning more than 200 individual sites, comprising four distinct ecosystem classifications (estuaries, sub-alpine lakes, rivers, and managed wetlands). Within each ecosystem, we compared fish Hg concentrations among replicated sub-habitats and also evaluated the influence of land use, landscape composition, and biogeochemical drivers on fish Hg concentrations. We found substantial variability in fish Hg concentrations among adjacent sub-habitats within ecosystems. In estuarine environments, fish Hg concentrations were 7.4x higher in seasonal-saline wetlands than adjacent tidal wetland habitats. In riverine alcoves, preliminary data suggest that fish Hg concentrations were 1.5x higher than in fishes from paired mainstem river habitat. Among managed wetland habitats, fish Hg concentrations in rice fields were 2x higher than those in managed seasonal wetlands that were subjected to identical wetting and drying patterns. Across ecosystems, dissolved organic carbon (DOC) concentrations in surface waters were consistently correlated with fish Hg concentrations, highlighting its importance in Hg methylation and transport processes. Yet, the strength and direction of the relationships varied among habitat types. For example, fish Hg concentrations were positively correlated with DOC concentrations in riverine environments, whereas we found a negative correlation in alpine lakes. Instead, the most important determinant of fish Hg concentrations in alpine lakes was conifer tree density within a

Shoal bass hybridization in the Chattahoochee River Basin near Atlanta, Georgia

USGS Publications Warehouse

Taylor, Andrew T.; Tringali, Michael D.; O'Rourke, Patrick M.; Long, James M.

2018-01-01

The shoal bass (Micropterus cataractae) is a sportfish endemic to the Apalachicola-Chattahoochee-Flint Basin of the southeastern United States. Introgression with several non-native congeners poses a pertinent threat to shoal bass conservation, particularly in the altered habitats of the Chattahoochee River. Our primary objective was to characterize hybridization in shoal bass populations near Atlanta, Georgia, including a population inhabiting Big Creek and another in the main stem Chattahoochee River below Morgan Falls Dam (MFD). A secondary objective was to examine the accuracy of phenotypic identifications below MFD based on a simplified suite of characters examined in the field. Fish were genotyped with 16 microsatellite DNA markers, and results demonstrated that at least four black bass species were involved in introgressive hybridization. Of 62 fish genotyped from Big Creek, 27% were pure shoal bass and 65% represented either F1 hybrids of shoal bass x smallmouth bass (M. dolomieu) or unidirectional backcrosses towards shoal bass. Of 29 fish genotyped below MFD and downstream at Cochran Shoals, 45% were pure shoal bass. Six hybrid shoal bass included both F1 hybrids and backcrosses with non-natives including Alabama bass (M. henshalli), spotted bass (M. punctulatus), and smallmouth bass. Shoal bass alleles comprised only 21% of the overall genomic composition in Big Creek and 31% below MFD (when combined with Cochran Shoals). Phenotypic identification below MFD resulted in an overall correct classification rate of 86% when discerning pure shoal bass from all other non-natives and hybrids. Results suggest that although these two shoal bass populations feature some of the highest introgression rates documented, only a fleeting opportunity may exist to conserve pure shoal bass in both populations. Continued supplemental stocking of pure shoal bass below MFD appears warranted to thwart increased admixture among multiple black bass taxa, and a similar stocking

Detection of a variable number of ribosomal DNA loci by fluorescent in situ hybridization in Populus species.

PubMed

Prado, E A; Faivre-Rampant, P; Schneider, C; Darmency, M A

1996-10-01

Fluorescent in situ hybridization (FISH) was applied to related Populus species (2n = 19) in order to detect rDNA loci. An interspecific variability in the number of hybridization sites was revealed using as probe an homologous 25S clone from Populus deltoides. The application of image analysis methods to measure fluorescence intensity of the hybridization signals has enabled us to characterize major and minor loci in the 18S-5.8S-25S rDNA. We identified one pair of such rDNA clusters in Populus alba; two pairs, one major and one minor, in both Populus nigra and P. deltoides; and three pairs in Populus balsamifera, (two major and one minor) and Populus euroamericana (one major and two minor). FISH results are in agreement with those based on RFLP analysis. The pBG13 probe containing 5S sequence from flax detected two separate clusters corresponding to the two size classes of units that coexist within 5S rDNA of most Populus species. Key words : Populus spp., fluorescent in situ hybridization, FISH, rDNA variability, image analysis.

Biotechnology applied to fish reproduction: tools for conservation.

PubMed

de Siqueira-Silva, Diógenes Henrique; Saito, Taiju; Dos Santos-Silva, Amanda Pereira; da Silva Costa, Raphael; Psenicka, Martin; Yasui, George Shigueki

2018-04-29

This review discusses the new biotechnological tools that are arising and promising for conservation and enhancement of fish production, mainly regarding the endangered and the most economically important species. Two main techniques, in particular, are available to avoid extinction of endangered fish species and to improve the production of commercial species. Germ cell transplantation technology includes a number of approaches that have been studied, such as the transplantation of embryo-to-embryo blastomere, embryo-to-embryo differentiated PGC, larvae to larvae and embryo differentiated PGC, transplantation of spermatogonia from adult to larvae or between adults, and oogonia transplantation. However, the success of germ cell transplantation relies on the prior sterilization of fish, which can be performed at different stages of fish species development by means of several protocols that have been tested in order to achieve the best approach to produce a sterile fish. Among them, fish hybridization and triploidization, germline gene knockdown, hyperthermia, and chemical treatment deserve attention based on important results achieved thus far. This review currently used technologies and knowledge about surrogate technology and fish sterilization, discussing the stronger and the weaker points of each approach.

Assessing connectivity of estuarine fishes based on stable isotope ratio analysis

NASA Astrophysics Data System (ADS)

Herzka, Sharon Z.

2005-07-01

Assessing connectivity is fundamental to understanding the population dynamics of fishes. I propose that isotopic analyses can greatly contribute to studies of connectivity in estuarine fishes due to the high diversity of isotopic signatures found among estuarine habitats and the fact that variations in isotopic composition at the base of a food web are reflected in the tissues of consumers. Isotopic analysis can be used for identifying nursery habitats and estimating their contribution to adult populations. If movement to a new habitat is accompanied by a shift to foods of distinct isotopic composition, recent immigrants and residents can be distinguished based on their isotopic ratios. Movement patterns thus can be reconstructed based on information obtained from individuals. A key consideration is the rate of isotopic turnover, which determines the length of time that an immigrant to a given habitat will be distinguishable from a longtime resident. A literature survey indicated that few studies have measured turnover rates in fishes and that these have focused on larvae and juveniles. These studies reveal that biomass gain is the primary process driving turnover rates, while metabolic turnover is either minimal or undetectable. Using a simple dilution model and biomass-specific growth rates, I estimated that young fishes with fast growth rates will reflect the isotopic composition of a new diet within days or weeks. Older or slower-growing individuals may take years or never fully equilibrate. Future studies should evaluate the factors that influence turnover rates in fishes during various stages of the life cycle and in different tissues, as well as explore the potential for combining stable isotope and otolith microstructure analyses to examine the relationship between demographic parameters, movement and connectivity.

Dispersal and selection mediate hybridization between a native and invasive species.

PubMed

Kovach, Ryan P; Muhlfeld, Clint C; Boyer, Matthew C; Lowe, Winsor H; Allendorf, Fred W; Luikart, Gordon

2015-01-22

Hybridization between native and non-native species has serious biological consequences, but our understanding of how dispersal and selection interact to influence invasive hybridization is limited. Here, we document the spread of genetic introgression between a native (Oncorhynchus clarkii) and invasive (Oncorhynchus mykiss) trout, and identify the mechanisms influencing genetic admixture. In two populations inhabiting contrasting environments, non-native admixture increased rapidly from 1984 to 2007 and was driven by surprisingly consistent processes. Individual admixture was related to two phenotypic traits associated with fitness: size at spawning and age of juvenile emigration. Fish with higher non-native admixture were larger and tended to emigrate at a younger age--relationships that are expected to confer fitness advantages to hybrid individuals. However, strong selection against non-native admixture was evident across streams and cohorts (mean selection coefficient against genotypes with non-native alleles (s) = 0.60; s.e. = 0.10). Nevertheless, hybridization was promoted in both streams by the continuous immigration of individuals with high levels of non-native admixture from other hybrid source populations. Thus, antagonistic relationships between dispersal and selection are mediating invasive hybridization between these fish, emphasizing that data on dispersal and natural selection are needed to fully understand the dynamics of introgression between native and non-native species. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Various Characteristics of Hybrid between River Puffer, Takifugu obscurus and Tiger Puffer, T. rubripes, and Their Hybrid Triploid

PubMed Central

Park, In-Seok; Lim, Sung Young; Lee, Tae Ho; Gil, Hyun Woo; Yoo, Gwang Yeol

2017-01-01

ABSTRACT A comparison of the growth, hematological values, fatty acids, and gonadal and growth hormonal changes of river puffer, Takifugu obscurus, tiger puffer, T. rubripes, their hybrids (river puffer × tiger puffer) and hybrid triploids was performed during 3 months of their early growth period. Several features were observed during these 3 months: hybrids showed the highest levels of specific growth rate, 1.48%; hybrid triploids showed the smallest change in viscera fat (P<0.05), but GSI was not significantly different among groups (P>0.05). Considering hematological parameters, hybrid triploids had increased mean corpuscular volume and mean corpuscular hemoglobin (P<0.05), but other parameters were not significantly different between groups (P>0.05). With respect to fatty acids, puffer fish, hybrids and hybrid triploids contained fatty acids such as SFAs, MUFAs, n-3 PUFAs and n-6 PUFAs. There were significantly different amounts of total fatty acids between groups (P<0.05), however, rates of changes in fatty acids did not differ significantly between groups (P>0.05). Gonadal hormone (estradiol and testosterone) changes in the river puffer and tiger puffer were significantly higher than that observed in hybrids and hybrid triploids. The hybrids and tiger puffers had higher amounts of growth hormone (thyroid stimulating hormone and thyroxine) than the hybrid triploids and river puffers (P<0.05). PMID:28785739

[Optimization and assessment of a reverse hybridization system for the detection of HBV drug-resistant mutations].

PubMed

Liu, Yan-chen; Huang, Ai-long; Hu, Yuan; Hu, Jie-li; Lai, Guo-qi; Zhang, Wen-lu

2011-12-01

To establish a detection method for HBV drug-resistant mutations related to lamivudine, adefovir and entecavir by optimization and assessment of reverse hybridization system. 26 degenerated probes covering 10 drug-resistant hotspots of 3 drugs were synthesized and immobilized on the same positively charged nylon membrane. PCR products labeled with digoxigenin were hybridized with corresponding probes. To improve the sensitivity and specificity, 4 reaction steps of reverse hybridization were optimized including the number of labeled digoxigenin, the energy intensity of UV cross-linking, hybridization and stringency wash conditions. To prove the feasibility, the specificity, sensitivity and accuracy of this system were assessed respectively. Sensitive and specific results are obtained by the optimization of the following 4 reaction steps: the primers labeled with 3 digoxigenin, energy intensity of UV cross-linking for 1500 x 0.1 mJ/cm², hybridization at 42 degrees C and stringency wash with 0.5 x SSC and 0.1% SDS solution at 44 degrees C for 30 min. In the assessment of system, the majority of probes have high specificity. The quantity of PCR product with a concentration of 10 ng/μl or above can be detected by this method. The concordant rate between reverse hybridization and direct sequencing is 93.9% in the clinical sample test. Though the specificity of several probes needs to be improved further, it is a simple, rapid and sensitive method which can detect HBV resistant mutations related to lamivudine, adefovir and entecavir simultaneously. Due to the short distance between 180 and 181, likewise 202 and 204, the sequence of the same probe covers two codon positions, and hybridization will be interfered by each other. To avoid such interference, the possible solution is that probes are designed by arranging and combining various forms of two near codons.

Assessing the potential for fish predation to impact zebra mussels (Dreissena polymorpha): Insight from bioenergetics models

USGS Publications Warehouse

Eggleton, M.A.; Miranda, L.E.; Kirk, J.P.

2004-01-01

Rates of annual food consumption and biomass were modeled for several fish species across representative rivers and lakes in eastern North America. Results were combined to assess the relative potential of fish predation to impact zebra mussels (Dreissena polymorpha). Predicted annual food consumption by fishes in southern waters was over 100% greater than that in northern systems because of warmer annual water temperatures and presumed increases in metabolic demand. Although generally increasing with latitude, biomasses of several key zebra mussel fish predators did not change significantly across latitudes. Biomasses of some less abundant fish predators did increase significantly with latitude, but increases were not of the magnitude to offset predicted decreases in food consumption. Our results generally support the premise that fishes in rivers and lakes of the southern United States (U.S.) have inherently greater potential to impact zebra mussels by predation. Our simulations may provide a partial explanation of why zebra mussel invasions have not been as rapid and widespread in southern U.S. waters compared to the Great Lakes region. ?? Blackwell Munksgaard, 2004.

Fluorescence In Situ Hybridization Probe Validation for Clinical Use.

PubMed

Gu, Jun; Smith, Janice L; Dowling, Patricia K

2017-01-01

In this chapter, we provide a systematic overview of the published guidelines and validation procedures for fluorescence in situ hybridization (FISH) probes for clinical diagnostic use. FISH probes-which are classified as molecular probes or analyte-specific reagents (ASRs)-have been extensively used in vitro for both clinical diagnosis and research. Most commercially available FISH probes in the United States are strictly regulated by the U.S. Food and Drug Administration (FDA), the Centers for Disease Control and Prevention (CDC), the Centers for Medicare & Medicaid Services (CMS) the Clinical Laboratory Improvement Amendments (CLIA), and the College of American Pathologists (CAP). Although home-brewed FISH probes-defined as probes made in-house or acquired from a source that does not supply them to other laboratories-are not regulated by these agencies, they too must undergo the same individual validation process prior to clinical use as their commercial counterparts. Validation of a FISH probe involves initial validation and ongoing verification of the test system. Initial validation includes assessment of a probe's technical specifications, establishment of its standard operational procedure (SOP), determination of its clinical sensitivity and specificity, development of its cutoff, baseline, and normal reference ranges, gathering of analytics, confirmation of its applicability to a specific research or clinical setting, testing of samples with or without the abnormalities that the probe is meant to detect, staff training, and report building. Ongoing verification of the test system involves testing additional normal and abnormal samples using the same method employed during the initial validation of the probe.

First among Equals: Hybridization of Cognitive Diagnostic Assessment and Evidence-Centered Game Design

ERIC Educational Resources Information Center

Leighton, Jacqueline P.; Chu, Man-Wai

2016-01-01

The objective of the present article is to explore differences and similarities between cognitive diagnostic assessment (CDA) and evidence-centered game design (ECgD) in the service of intentional hybridization. Although some testing specialists might argue that both are essentially the same given their origins in principled assessment design and…

Human health risk assessment of polycyclic aromatic hydrocarbons (PAHs) in smoked fish species from markets in Southern Nigeria.

PubMed

Tongo, Isioma; Ogbeide, Ozekeke; Ezemonye, Lawrence

2017-01-01

Polycyclic Aromatic Hydrocarbons (PAHs) levels in four commonly consumed smoked fish species from markets in Southern Nigeria were assessed to evaluate possible human health risks associated with consumption. Varying levels of PAH congeners were observed in the fish tissues with the highest total concentration of PAHs in Scomber scombrus . High concentrations of benzo(a)pyrene was observed in Clarias gariepinus and Ethmalosa fimbriata with values above the guideline value of 0.05 mg/kg. The Dietary Daily Intake (DDI) value for total PAHs (∑PAHs) was highest for S. scombrus while the DDI value for the total carcinogenic PAHs (∑CPAHs) was highest for E. fimbriata . Carcinogenic human health risk assessment using carcinogenic toxic equivalents (TEQ), indicated that consumption of E. fimbriata has a higher potential to cause carcinogenic risks. TEQ values for all the fish species were however, below the estimated screening value (SV) of 3.556 mg/kg, while the estimated cumulative excess cancer risk (ECR) for E. fimbriata and C. gariepinus and PAH4 index for all the assessed fish species exceeded threshold values indicating potential carcinogenic risk from consumption.

Evaluating and understanding fish health risks and their consequences in propagated and free-ranging fish populations

USGS Publications Warehouse

Moffitt, C.M.; Haukenes, A.H.; Williams, C.J.

2005-01-01

Fishery managers and resource conservationists are increasingly interested in understanding the fish health and disease risks of free-ranging fishes and whether propagated fishes or features and practices used at fish culture facilities pose a health risk to free-ranging populations. Disease agents are present in most both captive and all free-ranging fish populations, but the consequences and extent of infections in free-ranging populations are often difficult to measure, control, and understand. Sampling methods, protocols, and assay techniques developed to assess the health of captive populations are not as applicable for assessments of free-ranging fishes. The use of chemicals and therapeutics to control diseases and parasites in propagated fishes likely reduces the risk of introducing specific pathogens into the environment, but control measures may have localized effects on the environment surrounding fish culture facilities. To understand health risks of propagated and free ranging fishes, we must consider fish populations, culture facilities, fish releases, and their interactions within the greater geospatial features of the aquatic environment. ?? 2004 by the American Fisheries Society.

The fishes of Pea Ridge National Military Park, Arkansas, 2003

USGS Publications Warehouse

Justus, B.G.; Petersen, James C.

2005-01-01

A fish inventory was conducted at Pea Ridge National Military Park, Arkansas, during base-flow conditions in September 2003. Six sites including four streams and two ponds were sampled using conventional electrofishing equipment (a seine also was used at one site). There were 653 individuals collected comprising 18 species (plus 1 hybrid) and 15 genera. The number of species collected at the four stream sites ranged from 1 16. Most fish species collected generally are associated with small streams in the Ozark Plateaus. The two most common species were the banded sculpin and the southern redbelly dace. Three species and a sunfish hybrid were collected from the quarry pond. No fish were collected from the unnamed pond. A preliminary expected species list incorrectly listed 42 species because of incorrect species range or habitat requirements. One species not on the original list was added to the revised list. Upon revising this list, the inventory yielded 18 the 40 species (45 percent) and 1 hybrid. No previous fish inventories have been completed for park but some observations can be made relative to species distributions. There were only five fish species collected in three headwater streams, and it is unlikely that many other species would occur in these three streams because of constraints imposed on the fish community by stream size. Little Sugar Creek, a medium-sized stream, had the most species collected, and it is likely that additional species would be collected from this stream if additional sampling were to occur. Distribution records indicate that all 18 species occur in the general area. Although no species collected in this study are federallylisted threatened or endangered species, three species collected at Pea Ridge National Military Park may be of some special interest to National Park Service managers and others. Two the species collected (cardinal shiner and stippled darter) are endemic to the Ozark Plateaus; both are rather common in certain

Comparative toxicology for risk assessment of marine fishes and crustaceans. [Cyprinodon variegatus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Suter, G.W. II; Rosen, A.E.

1988-05-01

The goal of this study was to collect data on the effects of chemicals on marine fishes and crustaceans and to evaluate the predictive power of the data for assessing risks to marine resources. The data sets consisted of acute median lethal concentrations (LC{sub 50s}) and chronic maximum acceptable toxicant concentrations (MATCs). They were analyzed with regression models and simple comparisons. The conclusions include the following: (1) the variability found in the marine data was comparable to that found in freshwater data; (2) the standard marine test fish Cyprinodon variegatus appears to be representative of marine fishes; (3) the responsesmore » of marine crustaceans are so highly diverse that the concept of a representative crustacean is questionable; (4) mysid and penaeid shrimp appear to be particularly sensitive to toxic chemicals. These conclusions are subject to the constraints of the existing limited data base and should be confirmed by a systematic study of the relative sensitivity of marine organisms to chemicals with diverse modes of action.« less

Residual levels of rare earth elements in freshwater and marine fish and their health risk assessment from Shandong, China.

PubMed

Yang, Luping; Wang, Xining; Nie, Hongqian; Shao, Lijun; Wang, Guoling; Liu, Yongjun

2016-06-15

The total concentrations of rare earth elements (ΣREE) were quantified in 251 samples from 10 common species of freshwater and marine fish in seventeen cities of Shandong, China. ΣREE obtained from the freshwater fish ranged from 34.0 to 37.9ngg(-1) (wet weight) and marine fish from 12.7 to 37.6ngg(-1). The ratio of LREE to HREE was 13.7:1 and 10:1 for freshwater and marine fish, respectively. This suggests that freshwater fish exhibit greater REE concentrations than marine fish and the biological effects of LREE are higher than HREE. Results revealed a similar REE distribution pattern between those fish and coastal sediments, abiding the "abundance law". The health risk assessment demonstrated the EDIs of REEs in fish were significantly lower than the ADI, indicating that the consumption of these fish presents little risk to human health. Copyright © 2016 Elsevier Ltd. All rights reserved.

Thermodynamic framework to assess low abundance DNA mutation detection by hybridization

PubMed Central

Willems, Hanny; Jacobs, An; Hadiwikarta, Wahyu Wijaya; Venken, Tom; Valkenborg, Dirk; Van Roy, Nadine; Vandesompele, Jo; Hooyberghs, Jef

2017-01-01

The knowledge of genomic DNA variations in patient samples has a high and increasing value for human diagnostics in its broadest sense. Although many methods and sensors to detect or quantify these variations are available or under development, the number of underlying physico-chemical detection principles is limited. One of these principles is the hybridization of sample target DNA versus nucleic acid probes. We introduce a novel thermodynamics approach and develop a framework to exploit the specific detection capabilities of nucleic acid hybridization, using generic principles applicable to any platform. As a case study, we detect point mutations in the KRAS oncogene on a microarray platform. For the given platform and hybridization conditions, we demonstrate the multiplex detection capability of hybridization and assess the detection limit using thermodynamic considerations; DNA containing point mutations in a background of wild type sequences can be identified down to at least 1% relative concentration. In order to show the clinical relevance, the detection capabilities are confirmed on challenging formalin-fixed paraffin-embedded clinical tumor samples. This enzyme-free detection framework contains the accuracy and efficiency to screen for hundreds of mutations in a single run with many potential applications in molecular diagnostics and the field of personalised medicine. PMID:28542229

Thermodynamic framework to assess low abundance DNA mutation detection by hybridization.

PubMed

Willems, Hanny; Jacobs, An; Hadiwikarta, Wahyu Wijaya; Venken, Tom; Valkenborg, Dirk; Van Roy, Nadine; Vandesompele, Jo; Hooyberghs, Jef

2017-01-01

The knowledge of genomic DNA variations in patient samples has a high and increasing value for human diagnostics in its broadest sense. Although many methods and sensors to detect or quantify these variations are available or under development, the number of underlying physico-chemical detection principles is limited. One of these principles is the hybridization of sample target DNA versus nucleic acid probes. We introduce a novel thermodynamics approach and develop a framework to exploit the specific detection capabilities of nucleic acid hybridization, using generic principles applicable to any platform. As a case study, we detect point mutations in the KRAS oncogene on a microarray platform. For the given platform and hybridization conditions, we demonstrate the multiplex detection capability of hybridization and assess the detection limit using thermodynamic considerations; DNA containing point mutations in a background of wild type sequences can be identified down to at least 1% relative concentration. In order to show the clinical relevance, the detection capabilities are confirmed on challenging formalin-fixed paraffin-embedded clinical tumor samples. This enzyme-free detection framework contains the accuracy and efficiency to screen for hundreds of mutations in a single run with many potential applications in molecular diagnostics and the field of personalised medicine.

Potentials of Optical Damage Assessment Techniques in Automotive Crash-Concepts composed of FRP-Steel Hybrid Material Systems

NASA Astrophysics Data System (ADS)

Dlugosch, M.; Spiegelhalter, B.; Soot, T.; Lukaszewicz, D.; Fritsch, J.; Hiermaier, S.

2017-05-01

With car manufacturers simultaneously facing increasing passive safety and efficiency requirements, FRP-metal hybrid material systems are one way to design lightweight and crashworthy vehicle structures. Generic automotive hybrid structural concepts have been tested under crash loading conditions. In order to assess the state of overall damage and structural integrity, and primarily to validate simulation data, several NDT techniques have been assessed regarding their potential to detect common damage mechanisms in such hybrid systems. Significant potentials were found particularly in combining 3D-topography laser scanning and X-Ray imaging results. Ultrasonic testing proved to be limited by the signal coupling quality on damaged or curved surfaces.

USE OF SURROGATE SPECIES IN ASSESSING CONTAMINANT RISK TO ENDANGERED AND THREATENED FISHES

EPA Science Inventory

Surrogate species used in toxicity assessments must be carefully selected in order to be protective of listed species. At present, the rainbow trout is considered to be an acceptable surrogate for coldwater fishes. Similarly, the fathead minnow is considered to be an acceptable s...

Metrics for assessing freshwater fish in Narragansett Bay

EPA Science Inventory

Freshwater fish are ecologically important in stream ecosystems, and they provide significant value to humans. Historically, the streams and rivers of southern New England supported moderately diverse and abundant assemblages of native fishes. Currently, these habitats are impact...

Description and identification of American Black Duck, Mallard, and hybrid wing plumage

USGS Publications Warehouse

Kirby, R.E.; Reed, A.; Dupuis, P.; Obrecht, H.H.; Quist, W.J.

2000-01-01

We developed a key to identify wings of hybrids between American Black Ducks (Anas rubripes) and Mallards (A. platyrhynchos). Material for analysis included review of historical descriptions dating from the late 1700's, older museum collections in Europe and North America, wings collected from hunters in North America and Great Britain, birds banded in Canada and the United States, and a flock of propagated hybrids. All first filial generation (F1) American Black Duck x Mallard hybrids were identified correctly with the key. A lower proportion of other hybrid cohorts (i.e., backcrosses of F1, to parental forms (P1), and second and third filial generations (F2, F3, etc.) were identified. We successfully identified a larger portion of male than female hybrids for all hybrid progeny cohorts examined except F1. The new key identified 2.37 times more hybrids in the 1977 U.S. Fish and Wildlife Service Parts Collection Survey (annual determination of the species, age, and sex composition of the waterfowl harvest using detached wings contributed by hunters) than were identified by standard techniques. The proportion of American Black Duck x Mallard hybrids to the American Black Duck parental population (the ratio: hybrids/[hybrids + American Black Ducks]) may therefore be closer to 0.132 than 0.056, the historically reported value. The hybrid key is suggested for use from North Carolina north in the Atlantic Flyway and Arkansas and Tennessee north in the Mississippi Flyway (areas where other members of the Mallard group will not confound assessment). We provide suggestions for further research that would assist identification of wings in parts collection surveys and permit estimation of the proportional representation of Mallard genes in the American Black Duck gene pool.

Description and identification of American Black Duck, Mallard, and hybrid wing plumage

USGS Publications Warehouse

Kirby, Ronald E.; Reed, Austin; Dupuis, Pierre; Obrecht, Holliday H.; Quist, Walter J.

2000-01-01

We developed a key to identify wings of hybrids between American Black Ducks (Anas rubripes) and Mallards (A. platyrhynchos). Material for analysis included review of historical descriptions dating from the late 1700's, older museum collections in Europe and North America, wings collected from hunters in North America and Great Britain, birds banded in Canada and the United States, and a flock of propagated hybrids. All first filial generation (F1) American Black Duck - Mallard hybrids were identified correctly with the key. A lower proportion of other hybrid cohorts (i.e., backcrosses of F1 to parental forms (P1), and second and third filial generations (F2, F3, etc.) were identified. We successfully identified a larger portion of male than female hybrids for all hybrid progeny cohorts examined except F1. The new key identified 2.37 times more hybrids in the 1977 U.S. Fish and Wildlife Service Parts Collection Survey (annual determination of the species, age, and sex composition of the waterfowl harvest using detached wings contributed by hunters) than were identified by standard techniques. The proportion of American Black Duck - Mallard hybrids to the American Black Duck parental population (the ratio: hybrids/[hybrids + American Black Ducks]) may therefore be closer to 0.132 than 0.056, the historically reported value. The hybrid key is suggested for use from North Carolina north in the Atlantic Flyway and Arkansas and Tennessee north in the Mississippi Flyway (areas where other members of the Mallard group will not confound assessment). We provide suggestions for further research that would assist identification of wings in parts collection surveys and permit estimation of the proportional representation of Mallard genes in the American Black Duck gene pool.

A Rare Case of Klinefelter Syndrome Patient with Quintuple Mosaic Karyotype, Diagnosed by GTG-Banding and FISH

PubMed Central

Karimi, Hamideh; Sabbaghian, Marjan; Haratian, Kaveh; Vaziri Nasab, Hamed; Farrahi, Faramarz; Moradi, Shabnam Zari; Tavakolzadeh, Tayebeh; Beheshti, Zahra; Gourabi, Hamid; Meybodi, Anahita Mohseni

2014-01-01

Klinefelter syndrome (KS) is the most common sex chromosomal disorder in men. Most of these patients show the 47,XXY karyotype, whereas approximately 15% of them are mosaics with variable phenotype. A 39-year-old male investigated for primary infertility, was clinically normal with small firm testes and elevated levels of FSH, LH and low level of testosterone. Total azoospermia was confirmed on semen analysis. Testicular histopathology revealed no spermatogenesis and absence of germ cells. Karyotype from whole blood culture showed cells with 47,XXY/46,XX/ 45,X/48,XXXY/ 46,XY mosaicism. The predominant cell line was 47,XXY (83.67%). This was confirmed by fluorescence in situ hybridization (FISH). Also the presence of a small population of cells with the 48,XXXY and 45,X karyotypes was detected by FISH. This case illustrates the utility of FISH as an adjunct to conventional cytogenetics in assess the chromosome copy number in each cell line of a mosaic. PMID:25083188

A Rare Case of Klinefelter Syndrome Patient with Quintuple Mosaic Karyotype, Diagnosed by GTG-Banding and FISH.

PubMed

Karimi, Hamideh; Sabbaghian, Marjan; Haratian, Kaveh; Vaziri Nasab, Hamed; Farrahi, Faramarz; Moradi, Shabnam Zari; Tavakolzadeh, Tayebeh; Beheshti, Zahra; Gourabi, Hamid; Meybodi, Anahita Mohseni

2014-07-01

Klinefelter syndrome (KS) is the most common sex chromosomal disorder in men. Most of these patients show the 47,XXY karyotype, whereas approximately 15% of them are mosaics with variable phenotype. A 39-year-old male investigated for primary infertility, was clinically normal with small firm testes and elevated levels of FSH, LH and low level of testosterone. Total azoospermia was confirmed on semen analysis. Testicular histopathology revealed no spermatogenesis and absence of germ cells. Karyotype from whole blood culture showed cells with 47,XXY/46,XX/ 45,X/48,XXXY/ 46,XY mosaicism. The predominant cell line was 47,XXY (83.67%). This was confirmed by fluorescence in situ hybridization (FISH). Also the presence of a small population of cells with the 48,XXXY and 45,X karyotypes was detected by FISH. This case illustrates the utility of FISH as an adjunct to conventional cytogenetics in assess the chromosome copy number in each cell line of a mosaic.

Evaluation of hybrid neutralization/biosorption process for zinc ions removal from automotive battery effluent by dolomite and fish scales.

PubMed

Ribeiro, C; Scheufele, F B; Alves, H J; Kroumov, A D; Espinoza-Quiñones, F R; Módenes, A N; Borba, C E

2018-02-26

This work focused in the evaluation of Oreochromis niloticus fish scales (FS) as biosorbent material in the removal of Zn from a synthetic effluent based on automotive battery industry effluent and, further, a hybrid neutralization/biosorption process, aiming at a high-quality treated effluent, by a cooperative use of dolomite and FS. For this, a physicochemical and morphological characterization (i.e. SEM-EDX, FTIR, XRD, and TXRF) was performed, which helped to clarify a great heterogeneity of active sites (phosphate, carbonate, amide, and hydroxyl) on the biosorbent; also the inorganic constituents (apatites) leaching from the FS was identified. Biosorption results pointed out to a pH-dependent process due to changes in the functional group's anionic character (i.e. electrostatic interactions), where an initial pH = 3 favored the Zn uptake. Kinetic and equilibrium studies confirmed the heterogeneous surface and cooperative sorption, wherein experimental data were described by Generalized Elovich kinetic model and the favorable isotherm profile by Langmuir-Freundlich isotherm ([Formula: see text] = 15.38 mg g -1 and [Formula: see text]). Speciation diagram of Zn species along with the leached species demonstrated that, for the studied pH range, the biosorption was the most likely phenomena rather than precipitation. Finally, the hybrid neutralization/biosorption process showed great potential since both the Zn concentration levels and the pH reached the legislation standards (C Zn  = 4 mg L -1 ; pH = 5). Hence, based on the characterization and biosorption results, a comprehensive evaluation of the involved mechanisms in such complex system helped to verify the prospective of FS biosorbent for the Zn treatment from solution, in both individual and hybrid processes.

Effects of commonly used cooking practices on total mercury concentration in fish and their impact on exposure assessments.

PubMed

Morgan, J N; Berry, M R; Graves, R L

1997-01-01

The effects of cooking practices commonly used by Native Americans on total mercury concentrations in fish were investigated. A preparation factor relating mercury concentrations in fish as prepared for consumption to mercury concentration data as measured in typical environmental monitoring programs was calculated. Preparation factors are needed to provide risk assessors with a more accurate estimate of the actual amount of mercury ingested through consumption of contaminated fish. Data on fish preparation and consumption practices of two communities of Chippewa residing on the shores of Lake Superior in northern Wisconsin were used to select practices for study. The most commonly consumed species, walleye and lake trout, were selected. Whitefish livers were also selected for study. Commonly used cooking techniques including panfrying, deep-frying, baking, boiling, and smoking were duplicated in the laboratory. Total mercury concentrations were determined in fish portions before and after cooking and in a portion representative of that analyzed in programs to assess water quality (skin-on fillets). Total mercury was determined by microwave digestion-cold vapor atomic absorption spectroscopy. Mercury concentrations (wet weight basis) in panfried, baked, and boiled walleye fillets and deep-fried and baked whitefish livers ranged from 1.1 to 1.5 times higher than in corresponding raw portions. In lake trout, mercury concentrations were 1.5 to 2.0 times higher in cooked portions than in the raw portion. However, total mercury levels were constant before and after cooking, indicating the concentration effect is caused by weight (moisture and fat) loss. The addition of lemon juice to potentially release mercury from its bound state and promote volatilization did not exert any measurable influence on mercury concentrations in cooked walleye. In some cases mercury concentrations were increased with increased cooking times due to further loss of moisture and fat

Rapid Discrimination of Haemophilus influenzae, H. parainfluenzae, and H. haemolyticus by Fluorescence In Situ Hybridization (FISH) and Two Matrix-Assisted Laser-Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) Platforms

PubMed Central

Frickmann, Hagen; Christner, Martin; Donat, Martina; Berger, Anja; Essig, Andreas; Podbielski, Andreas; Hagen, Ralf Matthias; Poppert, Sven

2013-01-01

Background Due to considerable differences in pathogenicity, Haemophilus influenzae, H. parainfluenzae and H. haemolyticus have to be reliably discriminated in routine diagnostics. Retrospective analyses suggest frequent misidentifications of commensal H. haemolyticus as H. influenzae. In a multi-center approach, we assessed the suitability of fluorescence in situ hybridization (FISH) and matrix-assisted laser-desorption-ionization time-of-flight mass-spectrometry (MALDI-TOF-MS) for the identification of H. influenzae, H. parainfluenzae and H. haemolyticus to species level. Methodology A strain collection of 84 Haemophilus spp. comprising 50 H. influenzae, 25 H. parainfluenzae, 7 H. haemolyticus, and 2 H. parahaemolyticus including 77 clinical isolates was analyzed by FISH with newly designed DNA probes, and two different MALDI-TOF-MS systems (Bruker, Shimadzu) with and without prior formic acid extraction. Principal Findings Among the 84 Haemophilus strains analyzed, FISH led to 71 correct results (85%), 13 uninterpretable results (15%), and no misidentifications. Shimadzu MALDI-TOF-MS resulted in 59 correct identifications (70%), 19 uninterpretable results (23%), and 6 misidentifications (7%), using colony material applied directly. Bruker MALDI-TOF-MS with prior formic acid extraction led to 74 correct results (88%), 4 uninterpretable results (5%) and 6 misidentifications (7%). The Bruker MALDI-TOF-MS misidentifications could be resolved by the addition of a suitable H. haemolyticus reference spectrum to the system's database. In conclusion, no analyzed diagnostic procedure was free of errors. Diagnostic results have to be interpreted carefully and alternative tests should be applied in case of ambiguous test results on isolates from seriously ill patients. PMID:23646201

Mercury contamination in human hair and fish from Cambodia: levels, specific accumulation and risk assessment.

PubMed

Agusa, Tetsuro; Kunito, Takashi; Iwata, Hisato; Monirith, In; Tana, Touch Seang; Subramanian, Annamalai; Tanabe, Shinsuke

2005-03-01

Mercury (Hg) concentrations in human hair and fish samples from Phnom Penh, Kien Svay, Tomnup Rolork and Batrong, Cambodia, collected in November 1999 and December 2000 were determined to understand the status of contamination, and age- and sex-dependent accumulation in humans and to assess the intake of mercury via fish consumption. Mercury concentrations in human hair ranged from 0.54 to 190mug/g dry wt. About 3% of the samples contained Hg levels exceeding the no observed adverse effects level (NOAEL) of WHO (50mug/g) and the levels in some hair samples of women also exceeded the NOAEL (10mug/g) associated with fetus neurotoxicity. A weak but significant positive correlation was observed between age and Hg levels in hair of residents. Mercury concentrations in muscle of marine and freshwater fish from Cambodia ranged from <0.01 to 0.96mug/g wet wt. Mercury intake rates were estimated on the basis of the Hg content in fish and daily fish consumption. Three samples of marine fish including sharp-tooth snapper and obtuse barracuda, and one sample of sharp-tooth snapper exceeded the guidelines by US EPA and by Joint FAO/WHO Expert Committee on Food Additives (JECFA), respectively, which indicates that some fish specimens examined (9% and 3% for US EPA and JECFA guidelines, respectively) were hazardous for consumption at the ingestion rate of Cambodian people (32.6g/day). It is suggested that fish is probably the main source of Hg for Cambodian people. However, extremely high Hg concentrations were observed in some individuals and could not be explained by Hg intake from fish consumption, indicating some other contamination sources of Hg in Cambodia.

Assessing variation in the potential susceptibility of fish to pharmaceuticals, considering evolutionary differences in their physiology and ecology

PubMed Central

Brown, A. R.; Gunnarsson, L.; Kristiansson, E.; Tyler, C. R.

2014-01-01

Fish represent the planet's most diverse group of vertebrates and they can be exposed to a wide range of pharmaceuticals. For practical reasons, extrapolation of pharmaceutical effects from ‘model’ species to other fish species is adopted in risk assessment. Here, we critically assess this approach. First, we show that between 65% and 86% of human drug targets are evolutionarily conserved in 12 diverse fish species. Focusing on nuclear steroid hormone receptors, we further show that the sequence of the ligand binding domain that plays a key role in drug potency is highly conserved, but there is variation between species. This variation for the oestrogen receptor, however, does not obviously account for observed differences in receptor activation. Taking the synthetic oestrogen ethinyloestradiol as a test case, and using life-table-response experiments, we demonstrate significant reductions in population growth in fathead minnow and medaka, but not zebrafish, for environmentally relevant exposures. This finding contrasts with zebrafish being ranked as more ecologically susceptible, according to two independent life-history analyses. We conclude that while most drug targets are conserved in fish, evolutionary divergence in drug-target activation, physiology, behaviour and ecological life history make it difficult to predict population-level effects. This justifies the conventional use of at least a 10× assessment factor in pharmaceutical risk assessment, to account for differences in species susceptibility. PMID:25405965

ELEMENTAL FISH TISSUE CONTAMINATION IN NORTHEASTERN U.S. LAKES: EVALUATION OF AN APPROACH TO REGIONAL ASSESSMENT

EPA Science Inventory

The approach of the Environmental Monitoring and Assessment Program (EMAP) to monitoring of fish tissue contaminants is shown to have utility for regional assessment, and for discrimination of regional from local contamination. The survey sampling design employed EMAP can be used...

Progress in molecular diagnosis of Charcot-Marie-Tooth-disease type 1 (CMT 1, HMSN I) and hereditary neuropathy with liability to pressure palsies (HNPP) by fluorescence in situ hybridization (FISH)-detection of a potential genetic mosaicism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bathke, K.; Liehr. T.; Ekici, A.

1994-09-01

We tested 20 CMT 1 patients characterized according to the criteria of the European CMT consortium by Southern hybridization of MspI restricted genomic DNA with probes pVAW409R1, pVAW412Hec and pEW401HE. In 11 of the 20 CMT 1 cases (55%), we observed a duplication in 17q11.2; one patient had a dinucleotide insertion in exon 6 of the PO-gene (5%). One HNPP case had a typical 17p11.2 deletion. Analysis of CA-repeats was performed with primers RM11GT and Mfd41; SSCP-analysis of the PO, PMP22 and Cx32-genes is in progress. FISH was carried out with probe pVAW409R1. 125 interphase nuclei were analyzed for eachmore » proband by counting the signals per nucleus. Normal cells show a characteristic distribution of signals: 1 signal in 5.9% of nuclei, 2 in 86.3% and 3 in 7.8%. A duplication is indicated by a shift to 3 signals in more than approximately 60% and 2 in less than 25% of the nuclei. In contrast, the 17p11.2 deletion of the HNPP patient shifts to 82.4% of nuclei with a single hybridization signal versus 14.4% with 2 signals. We detected one case with significantly abnormal distribution of interphase nuclei hybridization signals compared to cultures of normal cells and to those with 17p11.2 duplication or deletion: 3.2% nuclei revealed 1 signal, 48.0% two signals and 48.8% 3 signals, indicating a pathogenic but moderate dosis increase compared to the throughout duplicated cases. FISH with probe pVAW409R1 is a versatile tool to detect the HNPP deletion both in interphase nuclei and in metaphase chromosomes. In CMT 1 disease interphase nuclei are required for FISH analysis due to the small duplication of 1.5 Mbp. In contrast to Southern techniques, FISH is able to detect genetic mosaicism.« less

Postponed feeding does not substantially reduce production expense during pond rearing of hybrid catfish fry

USDA-ARS?s Scientific Manuscript database

Production variables of hybrid catfish (ChannelCatfish Ictalurus punctatus×BlueCatfish I. furcatus) reared in nursery ponds and then stocked were compared between fish fed immediately after stocking (standard industry practice) and fish forwhich feedingwas postponed after stocking. Ponds (0.04 ha) w...

GeneCount: genome-wide calculation of absolute tumor DNA copy numbers from array comparative genomic hybridization data

PubMed Central

Lyng, Heidi; Lando, Malin; Brøvig, Runar S; Svendsrud, Debbie H; Johansen, Morten; Galteland, Eivind; Brustugun, Odd T; Meza-Zepeda, Leonardo A; Myklebost, Ola; Kristensen, Gunnar B; Hovig, Eivind; Stokke, Trond

2008-01-01

Absolute tumor DNA copy numbers can currently be achieved only on a single gene basis by using fluorescence in situ hybridization (FISH). We present GeneCount, a method for genome-wide calculation of absolute copy numbers from clinical array comparative genomic hybridization data. The tumor cell fraction is reliably estimated in the model. Data consistent with FISH results are achieved. We demonstrate significant improvements over existing methods for exploring gene dosages and intratumor copy number heterogeneity in cancers. PMID:18500990

Factors in exposure assessment: Ethnic and socio-economic differences in fishing and consumption of fish caught along the Savannah River

DOE Office of Scientific and Technical Information (OSTI.GOV)

Burger, J.; Stephens, W.L.; Boring, C.S.

1999-06-01

South Carolina has issued fish consumption advisories for the Savannah River based on mercury and radionuclide levels. The authors examine differences in fishing rates and fish consumption of 258 people interviewed while fishing along the Savannah River, as a function of age, education, ethnicity, employment history, and income, and test the assumption that the average consumption of fish is less than the recreational value of 19 kg/year assumed by risk assessors. Ethnicity and education contributed significantly to explaining variations in number of fish meals per month, serving size, and total quantity of fish consumed per year. Blacks fished more often,more » ate more fish meals of slightly larger serving sizes, and consumed more fish per year than did Whites. Although education and income were correlated, education contributed most significantly to behavior; people who did not graduate from high school ate fish more often, ate more fish per year, and ate more whole fish than people who graduated from high school. Computing consumption of fish for each person individually indicates that (1) people who eat fish more often also eat larger portions, (2) a substantial number of people consume more than the amount of fish used to compute risk to recreational fishermen, (3) some people consume more than the subsistence level default assumption (50 kg/year) and (4) Blacks consume more fish per year than Whites, putting them at greater risk from contaminants in fish. Overall, ethnicity, age, and education contributed to variations in fishing behavior and consumption.« less

ELEMENTAL FISH TISSUE CONTAMINATION IN NORTHEASTERN U.S. LAKES: EVALUATION OF AN APPROACH TO REGIONAL ASSESSMENT

EPA Science Inventory

The approach of the Environmental Monitoring and Assessment Program (EMAP) to monitoring of fish tissue contaminants is shown to have utility for regional assessment,and for discrimination of regional from local contamination.The survey sampling design employed by EMAP can be use...

Relative and combined effects of habitat and fishing on reef fish communities across a limited fishing gradient at Ningaloo.

PubMed

Wilson, Shaun K; Babcock, Russ C; Fisher, Rebecca; Holmes, Thomas H; Moore, James A Y; Thomson, Damian P

2012-10-01

Habitat degradation and fishing are major drivers of temporal and spatial changes in fish communities. The independent effects of these drivers are well documented, but the relative importance and interaction between fishing and habitat shifts is poorly understood, particularly in complex systems such as coral reefs. To assess the combined and relative effects of fishing and habitat we examined the composition of fish communities on patch reefs across a gradient of high to low structural complexity in fished and unfished areas of the Ningaloo Marine Park, Western Australia. Biomass and species richness of fish were positively correlated with structural complexity of reefs and negatively related to macroalgal cover. Total abundance of fish was also positively related to structural complexity, however this relationship was stronger on fished reefs than those where fishing is prohibited. The interaction between habitat condition and fishing pressure is primarily due to the high abundance of small bodied planktivorous fish on fished reefs. However, the influence of management zones on the abundance and biomass of predators and target species is small, implying spatial differences in fishing pressure are low and unlikely to be driving this interaction. Our results emphasise the importance of habitat in structuring reef fish communities on coral reefs especially when gradients in fishing pressure are low. The influence of fishing effort on this relationship may however become more important as fishing pressure increases. Copyright © 2012 Elsevier Ltd. All rights reserved.

Fluorescence in situ hybridization of TP53 for the detection of chromosome 17 abnormalities in myelodysplastic syndromes.

PubMed

Sánchez-Castro, Judit; Marco-Betés, Víctor; Gómez-Arbonés, Xavier; García-Cerecedo, Tomás; López, Ricard; Talavera, Elisabeth; Fernández-Ruiz, Sara; Ademà, Vera; Marugan, Isabel; Luño, Elisa; Sanzo, Carmen; Vallespí, Teresa; Arenillas, Leonor; Marco Buades, Josefa; Batlle, Ana; Buño, Ismael; Martín Ramos, María Luisa; Blázquez Rios, Beatriz; Collado Nieto, Rosa; Vargas, Ma Teresa; González Martínez, Teresa; Sanz, Guillermo; Solé, Francesc

2015-01-01

Conventional G-banding cytogenetics (CC) detects chromosome 17 (chr17) abnormalities in 2% of patients with de novo myelodysplastic syndromes (MDS). We used CC and fluorescence in situ hybridization (FISH) (LSI p53/17p13.1) to assess deletion of 17p in 531 patients with de novo MDS from the Spanish Group of Hematological Cytogenetics. FISH detected - 17 or 17p abnormalities in 13 cases (2.6%) in whom no 17p abnormalities were revealed by CC: 0.9% of patients with a normal karyotype, 0% in non-informative cytogenetics, 50% of patients with a chr17 abnormality without loss of 17p and 4.7% of cases with an abnormal karyotype not involving chr17. Our results suggest that applying FISH of 17p13 to identify the number of copies of the TP53 gene could be beneficial in patients with a complex karyotype. We recommend using FISH of 17p13 in young patients with a normal karyotype or non-informative cytogenetics, and always in isolated del(17p).

Pax1, a member of the paired box-containing class of developmental control genes, is mapped to human chromosome 20p11.2 by in situ hybridization (ISH and FISH).

PubMed

Schnittger, S; Rao, V V; Deutsch, U; Gruss, P; Balling, R; Hansmann, I

1992-11-01

Pax-1, a member of a murine multigene family, belongs to the paired box-containing class of developmental control genes first identified in Drosophila. The Pax-1 gene encodes a sequence-specific DNA-binding protein with transcriptional activating properties and has been found to be mutated in the autosomal recessive mutation undulated (un) on mouse chromosome 2 with vertebral anomalies along the entire rostrocaudal axis. By radioactive in situ hybridization (ISH) using a fragment from the murine Pax-1 paired box that is almost identical to the respective sequences from the cognate human gene HuP48 and fluorescence in situ hybridization (FISH) using a complete mouse Pax-1 cDNA, we have assigned the human homologue of murine Pax-1, the PAX1 locus, to chromosome 20p. The map position of PAX1 after FISH (FL-pter value of 0.34 +/- 0.04) corresponds to band p11.2. These results confirm the exceptional homology between human chromosome 20 and the distal segment of mouse chromosome 2, extending from bands F to G, and add PAX1 to the group of genes on 20p like PTPA, PRNP, SCG1, BMP2A, which are located in proximity on both chromosomes.

Flow Sorting of Marine Bacterioplankton after Fluorescence In Situ Hybridization

PubMed Central

Sekar, Raju; Fuchs, Bernhard M.; Amann, Rudolf; Pernthaler, Jakob

2004-01-01

We describe an approach to sort cells from coastal North Sea bacterioplankton by flow cytometry after in situ hybridization with rRNA-targeted horseradish peroxidase-labeled oligonucleotide probes and catalyzed fluorescent reporter deposition (CARD-FISH). In a sample from spring 2003 >90% of the cells were detected by CARD-FISH with a bacterial probe (EUB338). Approximately 30% of the microbial assemblage was affiliated with the Cytophaga-Flavobacterium lineage of the Bacteroidetes (CFB group) (probe CF319a), and almost 10% was targeted by a probe for the β-proteobacteria (probe BET42a). A protocol was optimized to detach cells hybridized with EUB338, BET42a, and CF319a from membrane filters (recovery rate, 70%) and to sort the cells by flow cytometry. The purity of sorted cells was >95%. 16S rRNA gene clone libraries were constructed from hybridized and sorted cells (S-EUB, S-BET, and S-CF libraries) and from unhybridized and unsorted cells (UNHYB library). Sequences related to the CFB group were significantly more frequent in the S-CF library (66%) than in the UNHYB library (13%). No enrichment of β-proteobacterial sequence types was found in the S-BET library, but novel sequences related to Nitrosospira were found exclusively in this library. These bacteria, together with members of marine clade OM43, represented >90% of the β-proteobacteria in the water sample, as determined by CARD-FISH with specific probes. This illustrates that a combination of CARD-FISH and flow sorting might be a powerful approach to study the diversity and potentially the activity and the genomes of different bacterial populations in aquatic habitats. PMID:15466568

Efficiency of fluorescence in situ hybridization for bacterial cell identification in temporary river sediments with contrasting water content.

PubMed

Fazi, Stefano; Amalfitano, Stefano; Pizzetti, Ilaria; Pernthaler, Jakob

2007-09-01

We studied the efficiency of two hybridization techniques for the analysis of benthic bacterial community composition under varying sediment water content. Microcosms were set up with sediments from four European temporary rivers. Wet sediments were dried, and dry sediments were artificially rewetted. The percentage of bacterial cells detected by fluorescence in situ hybridization with fluorescently monolabeled probes (FISH) significantly increased from dry to wet sediments, showing a positive correlation with the community activity measured via incorporation of (3)H leucine. FISH and signal amplification by catalyzed reporter deposition (CARD-FISH) could significantly better detect cells with low activity in dried sediments. Through the application of an optimized cell permeabilization protocol, the percentage of hybridized cells by CARD-FISH showed comparable values in dry and wet conditions. This approach was unrelated to (3)H leucine incorporation rates. Moreover, the optimized protocol allowed a significantly better visualization of Gram-positive Actinobacteria in the studied samples. CARD-FISH is, therefore, proposed as an effective technique to compare bacterial communities residing in sediments with contrasting water content, irrespective of differences in the activity state of target cells. Considering the increasing frequencies of flood and drought cycles in European temporary rivers, our approach may help to better understand the dynamics of microbial communities in such systems.

Wavelet-based compression of M-FISH images.

PubMed

Hua, Jianping; Xiong, Zixiang; Wu, Qiang; Castleman, Kenneth R

2005-05-01

Multiplex fluorescence in situ hybridization (M-FISH) is a recently developed technology that enables multi-color chromosome karyotyping for molecular cytogenetic analysis. Each M-FISH image set consists of a number of aligned images of the same chromosome specimen captured at different optical wavelength. This paper presents embedded M-FISH image coding (EMIC), where the foreground objects/chromosomes and the background objects/images are coded separately. We first apply critically sampled integer wavelet transforms to both the foreground and the background. We then use object-based bit-plane coding to compress each object and generate separate embedded bitstreams that allow continuous lossy-to-lossless compression of the foreground and the background. For efficient arithmetic coding of bit planes, we propose a method of designing an optimal context model that specifically exploits the statistical characteristics of M-FISH images in the wavelet domain. Our experiments show that EMIC achieves nearly twice as much compression as Lempel-Ziv-Welch coding. EMIC also performs much better than JPEG-LS and JPEG-2000 for lossless coding. The lossy performance of EMIC is significantly better than that of coding each M-FISH image with JPEG-2000.

Use of Hybridization Chain Reaction-Fluorescent In Situ Hybridization To Track Gene Expression by Both Partners during Initiation of Symbiosis.

PubMed

Nikolakakis, K; Lehnert, E; McFall-Ngai, M J; Ruby, E G

2015-07-01

The establishment of a productive symbiosis between Euprymna scolopes, the Hawaiian bobtail squid, and its luminous bacterial symbiont, Vibrio fischeri, is mediated by transcriptional changes in both partners. A key challenge to unraveling the steps required to successfully initiate this and many other symbiotic associations is characterization of the timing and location of these changes. We report on the adaptation of hybridization chain reaction-fluorescent in situ hybridization (HCR-FISH) to simultaneously probe the spatiotemporal regulation of targeted genes in both E. scolopes and V. fischeri. This method revealed localized, transcriptionally coregulated epithelial cells within the light organ that responded directly to the presence of bacterial cells while, at the same time, provided a sensitive means to directly show regulated gene expression within the symbiont population. Thus, HCR-FISH provides a new approach for characterizing habitat transition in bacteria and for discovering host tissue responses to colonization. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Mercury concentrations in fish from three major lakes in north Mississippi: Spatial and temporal differences and human health risk assessment.

PubMed

Wolff, Stacy; Brown, Garry; Chen, Jingjing; Meals, Keith; Thornton, Cammi; Brewer, Steve; Cizdziel, James V; Willett, Kristine L

2016-01-01

The goal of this study was to compare total mercury (Hg) concentrations in fish muscle tissue and assess consumption health risks of fish collected from three north Mississippi lakes (Sardis, Enid, and Grenada) that are extensively used for fishing and recreation. Largemouth bass (LMB; n = 64), channel catfish (CC; n = 72), and white crappie (WC; n = 100), which represent a range of trophic levels, were collected during spring 2013 and 2014. Creel data estimated that anglers harvested approximately 370,000 kg of WC, 27,000 kg of CC, and 15,000 kg of LMB from the lakes annually. Median Hg wet weight concentrations were highest in LMB (443 ng/g), followed by CC (211 ng/g) and WC (192 ng/g). Fish-Hg concentrations were lower than those reported in fish >10 years ago. There were significant differences between lakes consistent across species. Grenada length-normalized fish-Hg concentrations were higher than those from Enid and Sardis. Because existing consumption advisories for CC are length based, the lack of relationship between length and Hg concentration indicated that the recommendations may not be sufficiently protective. Further, five different risk assessment paradigms yielded hazard quotient (HQ) values suggesting that existing fish consumption advisories may be insufficient to protect adults and especially children from exposure to Hg.

Detecting Circular RNAs by RNA Fluorescence In Situ Hybridization.

PubMed

Zirkel, Anne; Papantonis, Argyris

2018-01-01

Fluorescence in situ hybridization (FISH) coupled to high-resolution microscopy is a powerful method for analyzing the subcellular localization of RNA. However, the detection of circular RNAs (circRNAs) using microscopy is challenging because the only feature of a circRNA that can be used for the probe design is its junction. Circular RNAs are expressed at varying levels, and for their efficient monitoring by FISH, background fluorescence levels need to be kept low. Here, we describe a FISH protocol coupled to high-precision localizations using a single fluorescently labeled probe spanning the circRNA junction; this allows circRNA detection in mammalian cells with high signal-to-noise ratios.

A FISH-based method for assessment of HER-2 amplification status in breast cancer circulating tumor cells following CellSearch isolation.

PubMed

Frithiof, Henrik; Aaltonen, Kristina; Rydén, Lisa

2016-01-01

Amplification of the HER-2/neu ( HER-2 ) proto-oncogene occurs in 10%-15% of primary breast cancer, leading to an activated HER-2 receptor, augmenting growth of cancer cells. Tumor classification is determined in primary tumor tissue and metastatic biopsies. However, malignant cells tend to alter their phenotype during disease progression. Circulating tumor cell (CTC) analysis may serve as an alternative to repeated biopsies. The Food and Drug Administration-approved CellSearch system allows determination of the HER-2 protein, but not of the HER-2 gene. The aim of this study was to optimize a fluorescence in situ hybridization (FISH)-based method to quantitatively determine HER-2 amplification in breast cancer CTCs following CellSearch-based isolation and verify the method in patient samples. Using healthy donor blood spiked with human epidermal growth factor receptor 2 (HER-2)-positive breast cancer cell lines, SKBr-3 and BT-474, and a corresponding negative control (the HER-2-negative MCF-7 cell line), an in vitro CTC model system was designed. Following isolation in the CellSearch system, CTC samples were further enriched and fixed on microscope slides. Immunocytochemical staining with cytokeratin and 4',6-diamidino-2'-phenylindole dihydrochloride identified CTCs under a fluorescence microscope. A FISH-based procedure was optimized by applying the HER2 IQFISH pharmDx assay for assessment of HER-2 amplification status in breast cancer CTCs. A method for defining the presence of HER-2 amplification in single breast cancer CTCs after CellSearch isolation was established using cell lines as positive and negative controls. The method was validated in blood from breast cancer patients showing that one out of six patients acquired CTC HER-2 amplification during treatment against metastatic disease. HER-2 amplification status of CTCs can be determined following CellSearch isolation and further enrichment. FISH is superior to protein assessment of HER-2 status in

Characterization and expression of Na+/K+-ATPase in gills and kidneys of the Teleost fish Oreochromis mossambicus, Oreochromis urolepis hornorum and their hybrids in response to salinity challenge.

PubMed

Zhu, Huaping; Liu, Zhigang; Gao, Fengying; Lu, Maixin; Liu, Yujiao; Su, Huanhuan; Ma, Dongmei; Ke, Xiaoli; Wang, Miao; Cao, Jianmeng; Yi, Mengmeng

2018-05-28

Tilapia (Oreochromis mossambicus, O. urolepis hornorum, their hybrids O. mossambicus♀ × O. hornorum♂ and O. hornorum♀ × O. mossambicus♂) were exposed to a high salinity environment to evaluate their osmoregulatory responses. The plasma osmolality of all the tilapia species were elevated with the salinity challenge. The activities of Na + /K + -ATPase (NKA) in both the gill and kidney showed a similar increased change tendency compared with the control. The distribution of NKA α1 mRNA in all the examined tissues suggested that NKA α1 has a possible housekeeping role for this isoform. The amount of NKA α1 mRNA in the gill and kidney was elevated in the four fishes with similar expression patterns after transfer from freshwater to seawater. The NKAα1 mRNA expression levels in the gill reached their peak level at 24 h after transfer (P < 0.01) compared to the freshwater group, following decreases in the pretreatment level at 48 h (P > 0.05). However, the NKAα1 mRNA expression levels in the kidney were not significantly affected with increasing environmental salinity (P > 0.05). The differences in the responses to saltwater challenge may be associated with differences in saltwater tolerance between the four tilapia. The drastic increase in the plasma osmolality, NKA activities and mRNA expression suggested that the hybrids (O. mossambicus♀ × O. hornorum♂) possess heterosis in salinity responsiveness compared to that of both the parents, indicating a maternal effect on the salinity tolerance of the tilapia hybrids. This study provides a theoretical basis to further study the mechanism of fish osmoregulation response to salinity challenge. Copyright © 2018 Elsevier Inc. All rights reserved.

A hybrid model to assess the impact of climate variability on streamflow for an ungauged mountainous basin

NASA Astrophysics Data System (ADS)

Wang, Chong; Xu, Jianhua; Chen, Yaning; Bai, Ling; Chen, Zhongsheng

2018-04-01

To quantitatively assess the impact of climate variability on streamflow in an ungauged mountainous basin is a difficult and challenging work. In this study, a hybrid model combing downscaling method based on earth data products, back propagation artificial neural networks (BPANN) and weights connection method was developed to explore an approach for solving this problem. To validate the applicability of the hybrid model, the Kumarik River and Toshkan River, two headwaters of the Aksu River, were employed to assess the impact of climate variability on streamflow by using this hybrid model. The conclusion is that the hybrid model presented a good performance, and the quantitative assessment results for the two headwaters are: (1) the precipitation respectively increased by 48.5 and 41.0 mm in the Kumarik catchment and Toshkan catchment, and the average annual temperature both increased by 0.1 °C in the two catchments during each decade from 1980 to 2012; (2) with the warming and wetting climate, the streamflow respectively increased 1.5 × 108 and 3.3 × 108 m3 per decade in the Kumarik River and the Toshkan River; and (3) the contribution of the temperature and precipitation to the streamflow, which were 64.01 ± 7.34, 35.99 ± 7.34 and 47.72 ± 8.10, 52.26 ± 8.10%, respectively in the Kumarik catchment and Toshkan catchment. Our study introduced a feasible hybrid model for the assessment of the impact of climate variability on streamflow, which can be used in the ungauged mountainous basin of Northwest China.

AN ASSESSMENT OF FLYWHEEL HIGH POWER ENERGY STORAGE TECHNOLOGY FOR HYBRID VEHICLES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hansen, James Gerald

2012-02-01

An assessment has been conducted for the DOE Vehicle Technologies Program to determine the state of the art of advanced flywheel high power energy storage systems to meet hybrid vehicle needs for high power energy storage and energy/power management. Flywheel systems can be implemented with either an electrical or a mechanical powertrain. The assessment elaborates upon flywheel rotor design issues of stress, materials and aspect ratio. Twelve organizations that produce flywheel systems submitted specifications for flywheel energy storage systems to meet minimum energy and power requirements for both light-duty and heavy-duty hybrid applications of interest to DOE. The most extensivemore » experience operating flywheel high power energy storage systems in heavy-duty and light-duty hybrid vehicles is in Europe. Recent advances in Europe in a number of vehicle racing venues and also in road car advanced evaluations are discussed. As a frame of reference, nominal weight and specific power for non-energy storage components of Toyota hybrid electric vehicles are summarized. The most effective utilization of flywheels is in providing high power while providing just enough energy storage to accomplish the power assist mission effectively. Flywheels are shown to meet or exceed the USABC power related goals (discharge power, regenerative power, specific power, power density, weight and volume) for HEV and EV batteries and ultracapacitors. The greatest technical challenge facing the developer of vehicular flywheel systems remains the issue of safety and containment. Flywheel safety issues must be addressed during the design and testing phases to ensure that production flywheel systems can be operated with adequately low risk.« less

A fine-scale assessment of using barriers to conserve native stream salmonids: a case study in Akokala Creek, Glacier National Park, USA

USGS Publications Warehouse

Muhlfeld, Clint C.; D'Angelo, Vincent S.; S. T. Kalinowski,; Landguth, Erin L.; C. C. Downs,; J. Tohtz,; Kershner, Jeffrey L.

2012-01-01

Biologists are often faced with the difficult decision in managing native salmonids of where and when to install barriers as a conservation action to prevent upstream invasion of nonnative fishes. However, fine-scale approaches to assess long-term persistence of populations within streams and watersheds chosen for isolation management are often lacking. We employed a spatially-explicit approach to evaluate stream habitat conditions, relative abundance, and genetic diversity of native westslope cutthroat trout (Oncorhynchus clarkii lewisi) within the Akokala Creek watershed in Glacier National Park- a population threatened by introgressive hybridization with nonnative rainbow trout (O. mykiss) from nearby sources. The systematic survey of 24 stream reaches showed broad overlap in fish population and suitable habitat characteristics among reaches and no natural barriers to fish migration were found. Analysis of population structure using 16 microsatellite loci showed modest amounts of genetic diversity among reaches, and that fish from Long Bow Creek were the only moderately distinct genetic group. We then used this information to assess the potential impacts of three barrier placement scenarios on long-term population persistence and genetic diversity. The two barrier placement scenarios in headwater areas generally failed to meet general persistence criteria for minimum population size (2,500 individuals, Ne = 500), maintenance of long-term genetic diversity (He), and no population subdivision. Conversely, placing a barrier near the stream mouth and selectively passing non-hybridized, migratory spawners entering Akokala Creek met all persistence criteria and may offer the best option to conserve native trout populations and life history diversity. Systematic, fine-scale stream habitat, fish distribution, and genetic assessments in streams chosen for barrier installation are needed in conjunction with broader scale assessments to understand the potential impacts of

Hybrid Wing Body Aircraft System Noise Assessment with Propulsion Airframe Aeroacoustic Experiments

NASA Technical Reports Server (NTRS)

Thomas, Russell H.; Burley, Casey L.; Olson, Erik D.

2010-01-01

A system noise assessment of a hybrid wing body configuration was performed using NASA s best available aircraft models, engine model, and system noise assessment method. A propulsion airframe aeroacoustic effects experimental database for key noise sources and interaction effects was used to provide data directly in the noise assessment where prediction methods are inadequate. NASA engine and aircraft system models were created to define the hybrid wing body aircraft concept as a twin engine aircraft with a 7500 nautical mile mission. The engines were modeled as existing technology high bypass ratio turbofans. The baseline hybrid wing body aircraft was assessed at 22 dB cumulative below the FAA Stage 4 certification level. To determine the potential for noise reduction with relatively near term technologies, seven other configurations were assessed beginning with moving the engines two fan nozzle diameters upstream of the trailing edge and then adding technologies for reduction of the highest noise sources. Aft radiated noise was expected to be the most challenging to reduce and, therefore, the experimental database focused on jet nozzle and pylon configurations that could reduce jet noise through a combination of source reduction and shielding effectiveness. The best configuration for reduction of jet noise used state-of-the-art technology chevrons with a pylon above the engine in the crown position. This configuration resulted in jet source noise reduction, favorable azimuthal directivity, and noise source relocation upstream where it is more effectively shielded by the limited airframe surface, and additional fan noise attenuation from acoustic liner on the crown pylon internal surfaces. Vertical and elevon surfaces were also assessed to add shielding area. The elevon deflection above the trailing edge showed some small additional noise reduction whereas vertical surfaces resulted in a slight noise increase. With the effects of the configurations from the

The IUCN Red List of Threatened Species: an assessment of coral reef fishes in the US Pacific Islands

NASA Astrophysics Data System (ADS)

Zgliczynski, B. J.; Williams, I. D.; Schroeder, R. E.; Nadon, M. O.; Richards, B. L.; Sandin, S. A.

2013-09-01

Widespread declines among many coral reef fisheries have led scientists and managers to become increasingly concerned over the extinction risk facing some species. To aid in assessing the extinction risks facing coral reef fishes, large-scale censuses of the abundance and distribution of individual species are critically important. We use fisheries-independent data collected as part of the NOAA Pacific Reef Assessment and Monitoring Program from 2000 to 2009 to describe the range and density across the US Pacific of coral reef fishes included on The International Union for the Conservation of Nature's (IUCN) 2011 Red List of Threatened Species. Forty-five species, including sharks, rays, groupers, humphead wrasse ( Cheilinus undulatus), and bumphead parrotfish ( Bolbometopon muricatum), included on the IUCN List, were recorded in the US Pacific Islands. Most species were generally rare in the US Pacific with the exception of a few species, principally small groupers and reef sharks. The greatest diversity and densities of IUCN-listed fishes were recorded at remote and uninhabited islands of the Pacific Remote Island Areas; in general, lower densities were observed at reefs of inhabited islands. Our findings complement IUCN assessment efforts, emphasize the efficacy of large-scale assessment and monitoring efforts in providing quantitative data on reef fish assemblages, and highlight the importance of protecting populations at remote and uninhabited islands where some species included on the IUCN Red List of Threatened Species can be observed in abundance.

A comparison between benthic gillnet and bottom trawl for assessing fish assemblages in a shallow eutrophic lake near the Changjiang River estuary

NASA Astrophysics Data System (ADS)

Li, Yalei; Liu, Qigen; Chen, Liping; Zhao, Liangjie; Wu, Hao; Chen, Liqiao; Hu, Zhongjun

2018-03-01

Two fishing methods including gillnetting and trawling to estimate attributes of fish assemblage were compared in Dianshan Lake from August 2009 to July 2010. Species composition differed significantly between the gears, with four significant contributors in gillnet catches and one in trawl catches. Trawling collected more proportions of benthic species by number and biomass than gillnetting. Size distribution was significantly influenced by fishing technique; gillnetting captured relatively less small-sized fishes and trawling captured less large-sized individuals. Trawling produced species richness closer to the one expected than gillnetting. On the whole, trawl catch was a quadratic polynomial function of gillnet catch and a significantly negative correlation was found between them, both of which varied as different polynomial functions of temperature. However, trawl and gillnet catches were significantly correlated only in one of five month groups. It is concluded that single-gear-based surveys can be misleading in assessments of attributes of fish assemblages, bottom trawling is a more effective gear for assessing fish diversity than benthic gillnetting, and using gillnet catches as an indicator of fish density depends on fishing season in the lake.

Towards Fluorescence In Vivo Hybridization (FIVH) Detection of H. pylori in Gastric Mucosa Using Advanced LNA Probes

PubMed Central

Fontenete, Sílvia; Leite, Marina; Guimarães, Nuno; Madureira, Pedro; Ferreira, Rui Manuel; Figueiredo, Céu; Wengel, Jesper; Azevedo, Nuno Filipe

2015-01-01

In recent years, there have been several attempts to improve the diagnosis of infection caused by Helicobacter pylori. Fluorescence in situ hybridization (FISH) is a commonly used technique to detect H. pylori infection but it requires biopsies from the stomach. Thus, the development of an in vivo FISH-based method (FIVH) that directly detects and allows the visualization of the bacterium within the human body would significantly reduce the time of analysis, allowing the diagnosis to be performed during endoscopy. In a previous study we designed and synthesized a phosphorothioate locked nucleic acid (LNA)/ 2’ O-methyl RNA (2’OMe) probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH). In conclusion, this methodology is a promising approach that might be used in vivo in the future in combination with a confocal laser endomicroscope for H. pylori visualization. PMID:25915865

FISHing for bacteria in food--a promising tool for the reliable detection of pathogenic bacteria?

PubMed

Rohde, Alexander; Hammerl, Jens Andre; Appel, Bernd; Dieckmann, Ralf; Al Dahouk, Sascha

2015-04-01

Foodborne pathogens cause millions of infections every year and are responsible for considerable economic losses worldwide. The current gold standard for the detection of bacterial pathogens in food is still the conventional cultivation following standardized and generally accepted protocols. However, these methods are time-consuming and do not provide fast information about food contaminations and thus are limited in their ability to protect consumers in time from potential microbial hazards. Fluorescence in situ hybridization (FISH) represents a rapid and highly specific technique for whole-cell detection. This review aims to summarize the current data on FISH-testing for the detection of pathogenic bacteria in different food matrices and to evaluate its suitability for the implementation in routine testing. In this context, the use of FISH in different matrices and their pretreatment will be presented, the sensitivity and specificity of FISH tests will be considered and the need for automation shall be discussed as well as the use of technological improvements to overcome current hurdles for a broad application in monitoring food safety. In addition, the overall economical feasibility will be assessed in a rough calculation of costs, and strengths and weaknesses of FISH are considered in comparison with traditional and well-established detection methods. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Life cycle assessment on microalgal biodiesel production using a hybrid cultivation system.

PubMed

Adesanya, Victoria O; Cadena, Erasmo; Scott, Stuart A; Smith, Alison G

2014-07-01

A life cycle assessment (LCA) was performed on a putative biodiesel production plant in which the freshwater alga Chlorella vulgaris, was grown using an existing system similar to a published commercial-scale hybrid cultivation. The hybrid system couples airlift tubular photobioreactors with raceway ponds in a two-stage process for high biomass growth and lipid accumulation. The results show that microalgal biodiesel production would have a significantly lower environmental impact than fossil-derived diesel. Based on the functional unit of 1 ton of biodiesel produced, the hybrid cultivation system and hypothetical downstream process (base case) would have 42% and 38% savings in global warming potential (GWP) and fossil-energy requirements (FER) when compared to fossil-derived diesel, respectively. Sensitivity analysis was performed to identify the most influential process parameters on the LCA results. The maximum reduction in GWP and FER was observed under mixotrophic growth conditions with savings of 76% and 75% when compared to conventional diesel, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

Simazine biodegradation in soil: analysis of bacterial community structure by in situ hybridization.

PubMed

Caracciolo, Anna Barra; Grenni, Paola; Ciccoli, Roberto; Di Landa, Giuseppe; Cremisini, Carlo

2005-09-01

Pesticide and nitrate contamination of soil and groundwater from agriculture is an environmental and public health concern worldwide. Simazine, 6-chloro-N2,N4-diethyl-1,3,5-triazine-2,4-diamine, is a triazine herbicide used in agriculture for selective weed control with several types of crops and it is frequently applied to soils receiving N-fertilizers. Degradation experiments were performed in the laboratory to assess whether the biodegradation of simazine in soil may be influenced by the presence of urea. Simazine degradation rates under different experimental conditions (presence/absence of urea, microbiologically active/sterilized soil) were assessed together with the formation, degradation and transformation of its main metabolites in soil. Simazine degradation was affected by the presence of urea, in terms both of a smaller half-life (t(1/2)) and of a higher amount of desethyl-simazine formed. The soil bacterial community was also studied. Microbial abundances were determined by epifluorescence direct counting. Moreover in situ hybridization with rRNA-targeted fluorescent oligonucleotide probes was used to analyze the bacterial community structure. Fluorescent in situ hybridization (FISH) was used to detect specific groups of bacteria such as the alpha,beta,gamma-subdivisions of Proteobacteria, Gram-positive bacteria with a high G + C DNA content, Planctomycetes, Betaproteobacterial ammonia-oxidizing bacteria and nitrifying bacteria. The presence of the herbicide and/or urea affected the bacterial community structure, showing that FISH is a valuable tool for determining the response of bacterial populations to different environmental conditions. Copyright 2005 Society of Chemical Industry

Assessing the New American Society of Clinical Oncology/College of American Pathologists Guidelines for HER2 Testing by Fluorescence In Situ Hybridization: Experience of an Academic Consultation Practice.

PubMed

Press, Michael F; Villalobos, Ivonne; Santiago, Angela; Guzman, Roberta; Cervantes, Monica; Gasparyan, Armen; Campeau, Anaamika; Ma, Yanling; Tsao-Wei, Denice D; Groshen, Susan

2016-04-15

Context .- Evaluation of HER2 gene amplification by fluorescence in situ hybridization (FISH) was changed by recent American Society of Clinical Oncology/College of American Pathologists (ASCO-CAP) guidelines. Objective . -To determine frequencies and assess patterns of HER2 protein expression for each ASCO-CAP guideline FISH category among 7526 breast cancers accrued to our consultation practice. Design .- We retrospectively reevaluated the HER2 FISH status of breast cancers in our consultation practice according to ASCO-CAP FISH guidelines, and documented HER2 protein levels in each category. Results . -According to new guidelines, 17.7% of our consultation breast cancers were "ISH-positive" with HER2:CEP17 FISH ratios ≥2.0 and average HER2 gene copies per cell ≥4.0 (group 1); 0.4% were "ISH-positive" with ratios ≥2.0 and average copies <4.0 (group 2); 0.6% were "ISH-positive" with ratios <2.0 and average copies ≥6.0 (group 3); 4.6% were "ISH-equivocal" with ratios <2.0 and average copies ≥4.0 and <6.0 (group 4); and 76.7% were "ISH-negative" with ratios <2.0 and average copies <4.0 (group 5). However, only groups 1 (HER2 amplified) and 5 (HER2 not amplified) agreed with our previously reported status, and only these groups demonstrated the expected immunohistochemistry status, overexpression and low expression, respectively. Groups 2 and 4 breast cancers lacked overexpression, whereas group 3 was not significantly associated with either increased or decreased HER2 expression. Conclusions .- Although the status of approximately 95% of our cases (groups 1 and 5) is not affected by the new guidelines, those of the other 5% (groups 2-4) conflict with previous HER2 gene amplification status and with HER2 status by immunohistochemistry.

A histology-based fish health assessment of the tigerfish, Hydrocynus vittatus from a DDT-affected area

NASA Astrophysics Data System (ADS)

McHugh, K. J.; Smit, N. J.; Van Vuren, J. H. J.; Van Dyk, J. C.; Bervoets, L.; Covaci, A.; Wepener, V.

The Pongolapoort Dam (PPD) in the Phongola River, Kwa-Zulu Natal, South Africa and the surrounding area are classified as intermediate to low risk malaria areas and are continually being treated with DDT for malaria vector control. DDT is known as an endocrine disrupting chemical posing estrogenic and anti-androgenic properties and therefore might impact on the health of the 18 freshwater fish species found within this system. Of these species the tigerfish, Hydrocynus vittatus, is targeted by both recreational and local subsistence fishermen and was recently included in the South African threatened or protected species list. Their protected status and importance as a food source therefore emphasises the need for their health status elucidation. Previous research on H. vittatus in the Phongola floodplain highlighted DDT biomagnification in this species. Recent data show that the sum of the DDT levels in February 2009 (5403.9 ng/g lipid) and July 2009 (5537.4 ng/g lipid) is still comparable to the high levels found 30 years earlier. The aim of the current study was thus to determine the health status of H. vittatus in relation to DDT exposure by means of a histology-based fish health assessment protocol. Tigerfish were collected in February 2009 ( n = 30) and July 2009 ( n = 15) and gill, kidney and liver tissue were subjected to histological analyses. Mean Index values showed that the Kidney Index ( IK), Gill Index ( IG) and Fish Index ( IFISH) were higher in fish from the February survey while the Liver Index was higher in those collected during July. Liver alterations identified included intercellular oedema, granular degeneration, vacuolation, nuclear pleomorphism and lymphocyte infiltration. Kidney alterations included dilation of the glomerulus capillaries, vacuolation and hyaline droplet degeneration. Gill alterations identified included telangiectasia and hyperplasia of the secondary lamella, congestion, and rupture of pillar cells. Although histological

Characterization of de novo duplications in eight patients by using fluorescence in situ hybridization with chromosome-specific DNA libraries

DOE Office of Scientific and Technical Information (OSTI.GOV)

Leana-Cox, J.; Wulfsberg, E.; Raffel, L.J.

Fluorescence in situ hybridization (FISH) with chromosome-specific DNA libraries was performed on samples from eight patients with de novo chromosomal duplications. In five cases, the clinical phenotype and/or cytogenetic evaluations suggested a likely origin of the duplicated material. In the remaining three cases, careful examination of the GTG-banding pattern indicated multiple possible origins; hybridization with more than one chromosome-specific library was performed on two of these cases. In all cases, FISH conclusively identified the chromosomal origin of the duplicated material. In addition, the hybridization pattern was useful in quantitatively delineating the duplication in two cases. 21 refs., 2 figs., 1more » tab.« less

Evaluation of telomere length in human cardiac tissues using cardiac quantitative FISH.

PubMed

Sharifi-Sanjani, Maryam; Meeker, Alan K; Mourkioti, Foteini

2017-09-01

Telomere length has been correlated with various diseases, including cardiovascular disease and cancer. The use of currently available telomere-length measurement techniques is often restricted by the requirement of a large amount of cells (Southern-based techniques) or the lack of information on individual cells or telomeres (PCR-based methods). Although several methods have been used to measure telomere length in tissues as a whole, the assessment of cell-type-specific telomere length provides valuable information on individual cell types. The development of fluorescence in situ hybridization (FISH) technologies enables the quantification of telomeres in individual chromosomes, but the use of these methods is dependent on the availability of isolated cells, which prevents their use with fixed archival samples. Here we describe an optimized quantitative FISH (Q-FISH) protocol for measuring telomere length that bypasses the previous limitations by avoiding contributions from undesired cell types. We have used this protocol on small paraffin-embedded cardiac-tissue samples. This protocol describes step-by-step procedures for tissue preparation, permeabilization, cardiac-tissue pretreatment and hybridization with a Cy3-labeled telomeric repeat complementing (CCCTAA) 3 peptide nucleic acid (PNA) probe coupled with cardiac-specific antibody staining. We also describe how to quantify telomere length by means of the fluorescence intensity and area of each telomere within individual nuclei. This protocol provides comparative cell-type-specific telomere-length measurements in relatively small human cardiac samples and offers an attractive technique to test hypotheses implicating telomere length in various cardiac pathologies. The current protocol (from tissue collection to image procurement) takes ∼28 h along with three overnight incubations. We anticipate that the protocol could be easily adapted for use on different tissue types.

Effects of fire on fish populations: Landscape perspectives on persistance of native fishes and nonnative fish invasions

USGS Publications Warehouse

Dunham, J.B.; Young, M.; Gresswell, Robert E.; Rieman, B.

2003-01-01

Our limited understanding of the short and long-term effects of fire on fish contributes to considerable uncertainty in assessments of the risks and benefits of fire management alternatives. A primary concern among the many potential effects of fire is the effects of fire and fire management on persistence of native fish populations. Limited evidence suggests vulnerability of fish to fire is contingent upon the quality of affected habitats, the amount and distribution of habitat (habitat fragmentation), and habitat specificity of the species in question. Species with narrow habitat requirements in highly degraded and fragmented systems are likely to be most vulnerable to fire and fire-related disturbance. In addition to effects of fire on native fish, there are growing concerns about the effects of fire on nonnative fish invasions. The role of fire in facilitating invasions by nonnative fishes is unknown, but experience with other species suggests some forms of disturbance associated with fire may facilitate invasion. Management efforts to promote persistence of fishes in fire-prone landscapes can take the form of four basic alternatives: (1) pre-fire management; (2) post-fire management; (3) managing fire itself (e.g. fire fighting); and (4) monitoring and adaptive management. Among these alternatives, pre-fire management is likely to be most effective. Effective pre-fire management activities will address factors that may render fish populations more vulnerable to the effects of fire (e.g. habitat degradation, fragmentation, and nonnative species). Post-fire management is also potentially important, but suffers from being a reactive approach that may not address threats in time to avert them. Managing fire itself can be important in some contexts, but negative consequences for fish populations are possible (e.g. toxicity of fire fighting chemicals to fish). Monitoring and adaptive management can provide important new information for evaluating alternatives, but

Evaluating analytical approaches for estimating pelagic fish biomass using simulated fish communities

USGS Publications Warehouse

Yule, Daniel L.; Adams, Jean V.; Warner, David M.; Hrabik, Thomas R.; Kocovsky, Patrick M.; Weidel, Brian C.; Rudstam, Lars G.; Sullivan, Patrick J.

2013-01-01

Pelagic fish assessments often combine large amounts of acoustic-based fish density data and limited midwater trawl information to estimate species-specific biomass density. We compared the accuracy of five apportionment methods for estimating pelagic fish biomass density using simulated communities with known fish numbers that mimic Lakes Superior, Michigan, and Ontario, representing a range of fish community complexities. Across all apportionment methods, the error in the estimated biomass generally declined with increasing effort, but methods that accounted for community composition changes with water column depth performed best. Correlations between trawl catch and the true species composition were highest when more fish were caught, highlighting the benefits of targeted trawling in locations of high fish density. Pelagic fish surveys should incorporate geographic and water column depth stratification in the survey design, use apportionment methods that account for species-specific depth differences, target midwater trawling effort in areas of high fish density, and include at least 15 midwater trawls. With relatively basic biological information, simulations of fish communities and sampling programs can optimize effort allocation and reduce error in biomass estimates.

USING A FISH INDEX TO ASSESS HABITAT QUALITY IN NARRAGANSETT BAY, RHODE ISLAND

EPA Science Inventory

We developed an estuarine index of biotic integrity to assess habitat quality in Narragansett Bay, Rhode Island. Fish were collected at 18 fixed stations with a 61-m x 3.05-m beach seine once per month in July and August from 1988 to 1999. Stations were designated high or low qua...

Effects of fire on fish populations: Landscape perspectives on persistence of native fishes and nonnative fish invasions

Treesearch

Jason B. Dunham; Michael K. Young; Robert E. Gresswell; Bruce E. Rieman

2003-01-01

Our limited understanding of the short and long-term effects of fire on fish contributes to considerable uncertainty in assessments of the risks and benefits of fire management alternatives. A primary concern among the many potential effects of fire is the effects of fire and fire management on persistence of native fish populations. Limited evidence suggests...

Preservation of large-scale chromatin structure in FISH experiments

PubMed Central

Hepperger, Claudia; Otten, Simone; von Hase, Johann

2006-01-01

The nuclear organization of specific endogenous chromatin regions can be investigated only by fluorescence in situ hybridization (FISH). One of the two fixation procedures is typically applied: (1) buffered formaldehyde or (2) hypotonic shock with methanol acetic acid fixation followed by dropping of nuclei on glass slides and air drying. In this study, we compared the effects of these two procedures and some variations on nuclear morphology and on FISH signals. We analyzed mouse erythroleukemia and mouse embryonic stem cells because their clusters of subcentromeric heterochromatin provide an easy means to assess preservation of chromatin. Qualitative and quantitative analyses revealed that formaldehyde fixation provided good preservation of large-scale chromatin structures, while classical methanol acetic acid fixation after hypotonic treatment severely impaired nuclear shape and led to disruption of chromosome territories, heterochromatin structures, and large transgene arrays. Our data show that such preparations do not faithfully reflect in vivo nuclear architecture. Electronic supplementary material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00412-006-0084-2 and is accessible for authorized users. PMID:17119992

Occurrence and human dietary assessment of fluoroquinolones antibiotics in cultured fish around tai lake, China.

PubMed

Song, Chao; Zhang, Cong; Kamira, Barry; Qiu, Liping; Fan, Limin; Wu, Wei; Meng, Shunlong; Hu, Gengdong; Chen, Jiazhang

2017-11-01

Fluoroquinolone antibiotics are widely used in the production of aquatic products and considered to be a significant contributing factor to the burden of both natural and aquaculture environments. However, the main types of fluoroquinolones present in aquaculture systems have not been determined. The objectives of the present study were to explore the occurrence of residual fluoroquinolone antibiotics in fish muscle tissues sampled from across the entire aquaculture season in the Tai Lake basin in China and to assess the dietary risks associated with the upcoming vendible fish in the last month of the aquaculture season. Fluoroquinolones were detected in 95.69% of all fish samples, and the concentrations ranged from the limit of quantification (LOQ) to 47 108.00 μg · kg -1 . Enrofloxacin contributed the most among the 9 fluoroquinolone antibiotics tested. Of the 4 fish species studied, enrofloxacin was present in bream at significant (p < 0.05) concentrations in August, with an average value of 321.45 μg · kg -1 , while enrofloxacin concentrations peaked in crab and shrimp in September, with average values of 6949.60 and 460.82 μg · kg -1 , respectively. However, different patterns were observed in perch, suggesting that other categories of antibiotics may be used in the production of this fish. Dietary risk assessment showed that the residual levels and estimated daily intake in bream, perch, and shrimp were far below the maximum residual level and acceptable daily intake. However, the average residual level in crab exceeded the maximum residual level, and the estimated daily intake value accounted for 78.49% of the acceptable daily intake, indicating potential consumption risk. In summary, the present study aims to guide the production and consumption of aquatic products. Environ Toxicol Chem 2017;36:2899-2905. © 2017 SETAC. © 2017 SETAC.

From Fishing to Fish Processing: Separation of Fish from Crustaceans in the Norway Lobster-Directed Multispecies Trawl Fishery Improves Seafood Quality

PubMed Central

Karlsen, Junita D.; Krag, Ludvig Ahm; Albertsen, Christoffer Moesgaard; Frandsen, Rikke Petri

2015-01-01

Fishing gears have negative impacts on seafood quality, especially on fish in the mixed trawl fishery targeting Norway lobster (Nephrops norvegicus). In this fishery, which is worth about €80 millions in Denmark alone, the quality of fish can be significantly improved by simple gear changes. A trawl codend divided into an upper and lower codend was designed to separate fish from Norway lobster during the fishing process by encourage fish to swim into the upper codend by using a frame at the entrance of the lower codend. Separate codends for fish and Norway lobster in the same gear provide the opportunity to selectively reduce small low-value fish, which will reduce catch weight and sorting time onboard the vessel. For this horizontally divided test codend and a standard codend, in which the catch was mixed, quality assessments were performed on the same batches of fish during three steps of the value chain: i) aboard the fishing vessel; ii) at the Fishermen’s Collection Central, and iii) in the production plant. Four species of fish and fillets from fish caught in the upper codend of the test codend were of significantly better quality for several of the assessed parameters compared with those caught in the standard codend: i) newly caught fish showed significantly less scale loss and discolourations and had significantly better texture; ii) landed fish had significantly better skin appearance and texture and significantly fewer discolourations; and iii) fillets showed significantly fewer blood spots and had significantly better texture. There were no differences in injuries for newly caught fish or gaping and bruises for fillets between the test and standard codends. The decrease in catch-related damages in the test codend is explained by little contact between fish and animals with hard or spiny surfaces due to successful separation of fish and Norway lobster into the upper and lower codends, respectively, and by lower catch weight in the upper codend of the

Genome profiling of ovarian adenocarcinomas using pangenomic BACs microarray comparative genomic hybridization

PubMed Central

Caserta, Donatella; Benkhalifa, Moncef; Baldi, Marina; Fiorentino, Francesco; Qumsiyeh, Mazin; Moscarini, Massimo

2008-01-01

Background Routine cytogenetic investigations for ovarian cancers are limited by culture failure and poor growth of cancer cells compared to normal cells. Fluorescence in situ Hybridization (FISH) application or classical comparative genome hybridization techniques are also have their own limitations in detecting genome imbalance especially for small changes that are not known ahead of time and for which FISH probes could not be thus designed. Methods We applied microarray comparative genomic hybridization (A-CGH) using one mega base BAC arrays to investigate chromosomal disorders in ovarian adenocarcinoma in patients with familial history. Results Our data on 10 cases of ovarian cancer revealed losses of 6q (4 cases mainly mosaic loss), 9p (4 cases), 10q (3 cases), 21q (3 cases), 22q (4 cases) with association to a monosomy X and gains of 8q and 9q (occurring together in 8 cases) and gain of 12p. There were other abnormalities such as loss of 17p that were noted in two profiles of the studied cases. Total or mosaic segmental gain of 2p, 3q, 4q, 7q and 13q were also observed. Seven of 10 patients were investigated by FISH to control array CGH results. The FISH data showed a concordance between the 2 methods. Conclusion The data suggest that A-CGH detects unique and common abnormalities with certain exceptions such as tetraploidy and balanced translocation, which may lead to understanding progression of genetic changes as well as aid in early diagnosis and have an impact on therapy and prognosis. PMID:18492273

Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF).

PubMed

Honeker, Linnea K; Root, Robert A; Chorover, Jon; Maier, Raina M

2016-12-01

Metal(loid)-contamination of the environment due to anthropogenic activities is a global problem. Understanding the fate of contaminants requires elucidation of biotic and abiotic factors that influence metal(loid) speciation from molecular to field scales. Improved methods are needed to assess micro-scale processes, such as those occurring at biogeochemical interfaces between plant tissues, microbial cells, and metal(loid)s. Here we present an advanced method that combines fluorescence in situ hybridization (FISH) with synchrotron-based multiple-energy micro-focused X-ray fluorescence microprobe imaging (ME μXRF) to examine colocalization of bacteria and metal(loid)s on root surfaces of plants used to phytostabilize metalliferous mine tailings. Bacteria were visualized on a small root section using SytoBC nucleic acid stain and FISH probes targeting the domain Bacteria and a specific group (Alphaproteobacteria, Gammaproteobacteria, or Actinobacteria). The same root region was then analyzed for elemental distribution and metal(loid) speciation of As and Fe using ME μXRF. The FISH and ME μXRF images were aligned using ImageJ software to correlate microbiological and geochemical results. Results from quantitative analysis of colocalization show a significantly higher fraction of As colocalized with Fe-oxide plaques on the root surfaces (fraction of overlap 0.49±0.19) than to bacteria (0.072±0.052) (p<0.05). Of the bacteria that colocalized with metal(loid)s, Actinobacteria, known for their metal tolerance, had a higher correlation with both As and Fe than Alphaproteobacteria or Gammaproteobacteria. This method demonstrates how coupling these micro-techniques can expand our understanding of micro-scale interactions between roots, metal(loid)s and microbes, information that should lead to improved mechanistic models of metal(loid) speciation and fate. Copyright © 2016 Elsevier B.V. All rights reserved.

A hybrid ARIMA and neural network model applied to forecast catch volumes of Selar crumenophthalmus

NASA Astrophysics Data System (ADS)

Aquino, Ronald L.; Alcantara, Nialle Loui Mar T.; Addawe, Rizavel C.

2017-11-01

The Selar crumenophthalmus with the English name big-eyed scad fish, locally known as matang-baka, is one of the fishes commonly caught along the waters of La Union, Philippines. The study deals with the forecasting of catch volumes of big-eyed scad fish for commercial consumption. The data used are quarterly caught volumes of big-eyed scad fish from 2002 to first quarter of 2017. This actual data is available from the open stat database published by the Philippine Statistics Authority (PSA)whose task is to collect, compiles, analyzes and publish information concerning different aspects of the Philippine setting. Autoregressive Integrated Moving Average (ARIMA) models, Artificial Neural Network (ANN) model and the Hybrid model consisting of ARIMA and ANN were developed to forecast catch volumes of big-eyed scad fish. Statistical errors such as Mean Absolute Errors (MAE) and Root Mean Square Errors (RMSE) were computed and compared to choose the most suitable model for forecasting the catch volume for the next few quarters. A comparison of the results of each model and corresponding statistical errors reveals that the hybrid model, ARIMA-ANN (2,1,2)(6:3:1), is the most suitable model to forecast the catch volumes of the big-eyed scad fish for the next few quarters.

Development of a spatially distributed model of fish population density for habitat assessment of rivers

NASA Astrophysics Data System (ADS)

Sui, Pengzhe; Iwasaki, Akito; Ryo, Masahiro; Saavedra, Oliver; Yoshimura, Chihiro

2013-04-01

Flow conditions play an important role in sustaining biodiversity of river ecosystem. However, their relations to freshwater fishes, especially to fish population density, have not been clearly described. This study, therefore, aimed to propose a new methodology to quantitatively link habitat conditions, including flow conditions and other physical conditions, to population density of fish species. We developed a basin-scale fish distribution model by integrating the concept of habitat suitability assessment with a distributed hydrological model (DHM) in order to estimate fish population density with particular attention to flow conditions. Generalized linear model (GLM) was employed to evaluate the relationship between population density of fish species and major environmental factors. The target basin was Sagami River in central Japan, where the river reach was divided into 10 sections by estuary, confluences of tributaries, and river-crossing structures (dams, weirs). The DHM was employed to simulate river discharge from 1998 to 2005, which was used to calculate 10 flow indices including mean discharge, 25th and 75th percentile discharge, duration of low and high flows, number of floods. In addition, 5 water quality parameters and 13 other physical conditions (such as basin area, river width, mean diameter of riverbed material, and number of river-crossing structures upstream and downstream) of each river section were considered as environmental variables. In case of Sagami River, 10 habitat variables among them were then selected based on their correlations to avoid multicollinearity. Finally, the best GLM was developed for each species based on Akaike's information criterion. As results, population densities of 16 fish species in Sagami River were modelled, and correlation coefficients between observed and calculated population densities for 10 species were more than 0.70. The key habitat factors for population density varied among fish species. Minimum

Assessing Potential Conservation and Restoration Areas of Freshwater Fish Fauna in the Indian River Basins

NASA Astrophysics Data System (ADS)

Bhatt, Jay P.; Manish, Kumar; Mehta, Rajender; Pandit, Maharaj K.

2016-05-01

Conservation efforts globally are skewed toward terrestrial ecosystems. To date, conservation of aquatic ecosystems, in particular fish fauna, is largely neglected. We provide a country-wide assessment of Indian river ecosystems in order to identify and prioritize areas for protection and restoration of freshwater fish fauna. Using various biodiversity and anthropogenic attributes, coupled with tools of ecological modeling, we delineated areas for fish fauna conservation and restoration in the 20 major river basins of India. To do this, we used prioritization analyses and reserve selection algorithms to derive conservation value index (CVI) and vulnerability index (VI) of the river basins. CVI was estimated using endemicity, rarity, conservation value, and taxonomic singularity, while VI was estimated using a disturbance index derived from percent geographic area of the basin under human settlements, human population density, predominant land use, and total number of exotic fish species in each basin. The two indices, CVI and VI, were converted into geo-referenced maps, and each map was super-imposed onto species richness and forest cover maps, respectively. After superimposition, areas with high CVI and low VI shade intensities were delineated for conservation, while areas with high CVI and high VI shade intensities were demarcated for restoration. In view of the importance of freshwater fish for human livelihoods and consumption, and ecosystems of India's rivers, we call for urgent attention to the conservation of their fish fauna along with restoration of their degraded habitats.

Automation of ALK gene rearrangement testing with fluorescence in situ hybridization (FISH): a feasibility study.

PubMed

Zwaenepoel, Karen; Merkle, Dennis; Cabillic, Florian; Berg, Erica; Belaud-Rotureau, Marc-Antoine; Grazioli, Vittorio; Herelle, Olga; Hummel, Michael; Le Calve, Michele; Lenze, Dido; Mende, Stefanie; Pauwels, Patrick; Quilichini, Benoit; Repetti, Elena

2015-02-01

In the past several years we have observed a significant increase in our understanding of molecular mechanisms that drive lung cancer. Specifically in the non-small cell lung cancer sub-types, ALK gene rearrangements represent a sub-group of tumors that are targetable by the tyrosine kinase inhibitor Crizotinib, resulting in significant reductions in tumor burden. Phase II and III clinical trials were performed using an ALK break-apart FISH probe kit, making FISH the gold standard for identifying ALK rearrangements in patients. FISH is often considered a labor and cost intensive molecular technique, and in this study we aimed to demonstrate feasibility for automation of ALK FISH testing, to improve laboratory workflow and ease of testing. This involved automation of the pre-treatment steps of the ALK assay using various protocols on the VP 2000 instrument, and facilitating automated scanning of the fluorescent FISH specimens for simplified enumeration on various backend scanning and analysis systems. The results indicated that ALK FISH can be automated. Significantly, both the Ikoniscope and BioView system of automated FISH scanning and analysis systems provided a robust analysis algorithm to define ALK rearrangements. In addition, the BioView system facilitated consultation of difficult cases via the internet. Copyright © 2015 Elsevier Inc. All rights reserved.

Novel expression of the stanniocalcin gene in fish.

PubMed

McCudden, C R; Kogon, M R; DiMattia, G E; Wagner, G F

2001-10-01

It is currently accepted that the fish stanniocalcin (STC) gene is expressed exclusively in the corpuscles of Stannius (CS), unique endocrine glands on the kidneys of bony fishes. In this study, we have re-examined the pattern of fish STC gene expression in the light of the recent evidence for widespread expression of the gene in mammals. Surprisingly, we found by Northern blotting that the fish gene was also expressed in the kidneys and gonads, in addition to the CS glands. Moreover, Southern blotting of RT-PCR products revealed STC mRNA transcripts in all tissues assayed, including brain, heart, gill, muscle and intestine. In situ hybridization studies using digoxigenin-labeled riboprobes localized STC mRNA to chondrocytes, and both mature and developing nephritic tubules. Immunocytochemical staining indicated that the STC protein was widespread in cells of the gill, kidney, brain, eye, pseudobranch and skin. We also characterized the salmon STC gene, establishing that it was comprised of five exons as opposed to four in mammals. A single transcription start site was identified by primer extension 99 bp upstream of the start codon. This is the first evidence of STC gene expression in fish tissues other than the CS glands and suggests that, as in mammals, fish STC operates via both local and endocrine pathways.

Towards an alternative for the acute fish LC(50) test in chemical assessment: the fish embryo toxicity test goes multi-species -- an update.

PubMed

Braunbeck, Thomas; Boettcher, Melanie; Hollert, Henner; Kosmehl, Thomas; Lammer, Eva; Leist, Erik; Rudolf, Mark; Seitz, Nadja

2005-01-01

After its standardisation at the national level in Germany (DIN 38415-6, 2001, 2001), the 48 h sewage testing assay with zebrafish (Danio rerio) embryos has been submitted for standardisation to ISO. As an alternative to the conventional acute (96 h) fish test, a modified fish embryo test will be submitted to the OECD for chemical testing in late 2005. For this, a protocol originally designed for zebrafish was adapted to fit also the requirements of other OECD species, namely medaka (Oryzias latipes) and fathead minnow (Pimephales promelas). Results document that the transfer of the protocol is possible with only minor modifications. Data obtained from embryo tests with the three species are comparable. Statistical analysis of existing zebrafish embryo toxicity data resulted in the conclusions (1) that there is a reliable correlation between the fish embryo test and the acute fish test, (2) that the confidence belt of the regression line was relatively small, but that the prediction range was relatively wide. The regression thus seems appropriate to describe the relationship between acute fish and embryo LC(50) with good confidence, but is less appropriate as a prediction model. Investigations into oxygen requirements of zebrafish embryos reveal that they adapt to a broad range of oxygen levels and survive at concentrations of 2 mg/l without malformations. Zebrafish embryos can thus be exposed in very small toxicant volumes (100 microl), which is of particular interest for the testing of metabolites. Dechorionation studies with 48 h old zebrafish embryos indicate that the barrier function of the chorion increases with the lipophilicity of the test compound. Finally, examples are given as to how additional endpoints can be incorporated into the fish embryo test protocol to extend its scope, e.g. to sediment toxicity assessment or genotoxicity and mutagenicity testing.

Assessing impacts of land-applied manure from concentrated animal feeding operations on fish populations and communities.

PubMed

Leet, Jessica K; Lee, Linda S; Gall, Heather E; Goforth, Reuben R; Sassman, Stephen; Gordon, Denise A; Lazorchak, James M; Smith, Mark E; Jafvert, Chad T; Javfert, Chad T; Sepúlveda, Maria S

2012-12-18

Concentrated animal feeding operation (CAFO) manure is a cost-effective fertilizer. In the Midwest, networks of subsurface tile-drains expedite transport of animal hormones and nutrients from land-applied CAFO manure to adjacent waterways. The objective of this study was to evaluate impacts of land-applied CAFO manure on fish populations and communities. Water chemistry including hormone, pesticide, and nutrient concentrations was characterized from study sites along with fish assemblage structure, growth, and endocrine disruption assessed in selected fish species. Although most CAFO water samples had hormone concentrations <1 ng/L, equivalent concentrations for 17β-E2 and 17α-TB peaked at >30 ng/L each during the period of spawning, hatching, and development for resident fishes. CAFO sites had lower fish species richness, and fishes exhibited faster somatic growth and lower reproductive condition compared to individuals from the reference site. Fathead minnows (Pimephales promelas) exposed to CAFO ditchwater during early developmental stages exhibited significantly skewed sex ratios toward males. Maximum observed hormone concentrations were well above the lowest observable effect concentrations for these hormones; however, complexities at the field scale make it difficult to directly relate hormone concentration and impacts on fish. Complicating factors include the consistent presence of pesticides and nutrients, and the difference in temperature and stream architecture of the CAFO-impacted ditches compared to the reference site (e.g., channelization, bottom substrate, shallow pools, and riparian cover).

[Rapid ecological assessment of tropical fish communities in a gold mine area of Costa Rica].

PubMed

Espinoza Mendiola, Mario

2008-12-01

Gold mining impacts have generated a great concern regarding aquatic systems and habitat fragmentation. Anthropogenic disturbances on the structure and heterogeneity of a system can have an important effect on aquatic community stability. Ecological rapid assessments (1996, 2002, and 2007) were employed to determine the structure, composition and distribution of tropical fish communities in several rivers and smaller creeks from a gold mining area in Cerro Crucitas, Costa Rica. In addition, species composition and relative abundance were related with habitat structure. A total of 35 species were registered, among which sardine Astyanax aeneus (Characidae) and livebearer Alfaro cultratus (Poeciliidae) were the most abundant fish (71%). The highest species richness was observed in Caño Crucitas (s=19) and Minas Creek (s=18). Significant differences in fish communities structure and composition from Infiernillo river and Minas creek were observed (lamda = 0.0, F(132, 66) = 2.24, p < 0.001). Presence and/or absence of certain species such as Dormitor gobiomorus, Rhamdia nicaraguensis, Parachromis loiseillei and Atractosteus tropicus explained most of the spatial variation among sites. Habitat structure also contributed to explain differences among sites (lamda = 0.004, F(60.183) = 5.52, p < 0.001). Substratum (soft and hard bottom types) and habitat attributes (elevation, width and depth) explained most of the variability observed in Infiernillo River, Caño Crucitas and Tamagá Creek. In addition, a significant association between fish species and habitat structure was observed. This study reveals a high complexity in tropical fish communities that inhabit a gold mine area. Furthermore, it highlights the importance of habitat heterogeneity in fish community dynamics. The loss and degradation of aquatic systems in Cerro Crucitas can have a strong negative effect on fish community structure and composition of local species. A better understanding of the use of specific

Assessment of fish communities in a Mediterranean MPA: Can a seasonal no-take zone provide effective protection?

NASA Astrophysics Data System (ADS)

Dimitriadis, Charalampos; Sini, Maria; Trygonis, Vasilis; Gerovasileiou, Vasilis; Sourbès, Laurent; Koutsoubas, Drosos

2018-07-01

The efficacy of a Mediterranean Marine Protected Area (National Marine Park of Zakynthos - NMPZ, Ionian Sea, Greece) that implements a seasonal no-take zone as part of its management scheme was assessed using fish data collected in situ with underwater visual census. Sampling was conducted at two habitat types (Posidonia oceanica meadows and rocky reefs) that occur at sites of different protection level with respect to fisheries (high protection: seasonal no-take zone within the MPA; intermediate: zones within the MPA where small-scale fishing is allowed; none: areas outside the MPA, where all types of fishing are allowed, including trawlers, purse seiners, and recreational fishing). The data were used to examine the effects of protection level and habitat type on community parameters, trophic structure and functional diversity of fish populations that occupy the upper sublittoral zone. Overall, habitat type had a more pronounced effect than protection level on all investigated parameters. Biomass, density and number of fish species with low commercial value were higher in sites of intermediate protection, but no substantial fisheries-related ecological benefits were detected for targeted fish in the seasonal no-take zone. Conducted 8 years after the initial implementation of the seasonal no-take management scheme, our study suggests that existing fishing regulations in the NMPZ provide some measurable effects, but fall short of maintaining sufficient protection for the recovery of apex predators or other commercially important fish species. A revision of the existing zoning system to include permanent no-take zones, alongside the regulation of professional fishing and all extractive activities in the rest of the MPA, are strongly encouraged in order to enhance the effectiveness of fisheries management.

Assessing the utility of eDNA as a tool to survey reef-fish communities in the Red Sea

NASA Astrophysics Data System (ADS)

DiBattista, Joseph D.; Coker, Darren J.; Sinclair-Taylor, Tane H.; Stat, Michael; Berumen, Michael L.; Bunce, Michael

2017-12-01

Relatively small volumes of water may contain sufficient environmental DNA (eDNA) to detect target aquatic organisms via genetic sequencing. We therefore assessed the utility of eDNA to document the diversity of coral reef fishes in the central Red Sea. DNA from seawater samples was extracted, amplified using fish-specific 16S mitochondrial DNA primers, and sequenced using a metabarcoding workflow. DNA sequences were assigned to taxa using available genetic repositories or custom genetic databases generated from reference fishes. Our approach revealed a diversity of conspicuous, cryptobenthic, and commercially relevant reef fish at the genus level, with select genera in the family Labridae over-represented. Our approach, however, failed to capture a significant fraction of the fish fauna known to inhabit the Red Sea, which we attribute to limited spatial sampling, amplification stochasticity, and an apparent lack of sequencing depth. Given an increase in fish species descriptions, completeness of taxonomic checklists, and improvement in species-level assignment with custom genetic databases as shown here, we suggest that the Red Sea region may be ideal for further testing of the eDNA approach.

Assessment of metal and bacterial contamination in cultivated fish and impact on human health for residents living in the Mekong Delta.

PubMed

Chanpiwat, Penradee; Sthiannopkao, Suthipong; Widmer, Kenneth; Himeno, Seiichiro; Miyataka, Hideki; Vu, Ngoc-Ut; Tran, Van-Viet; Pham, Thi-Tuyet-Ngan

2016-11-01

Fish is the main source of animal protein and micronutrients for inhabitants in the lower Mekong River basin. Consumption of fish in the basin ranges from 41 to 51 kg capita(-1) year(-1). Thus, concerns of human health impacts caused by daily intake of metals contained in fish, and the incidence of bacterial contamination from Listeria and Escherichia coli have been raised. This study was conducted to 1) determine concentrations of metals, fecal indicator organisms, and Listeria spp. in cultivated common diet fish, and 2) assess human health risks as results of fish consumption on a daily basis. The results showed significant impacts of metal accumulation in fish especially from the intensive aquaculture. Chemical use to promote the rapid allometric growth of fish was expected to be the explanation for this finding. Concentrations of metals contained in different fish species were not statistically different with the exceptions of Na, Mn, and Zn. This might be due to the mobility of elements in aquaculture farms. Listeria and E. coli log CFU/g were 1.36 ± 0.11 (standard error) and 1.57 ± 0.1 s.e., respectively with higher counts observed in samples collected in market sites. Lastly, for human health risk assessment via fish consumption, it was found that hazard quotients of consuming As, Cu, and Zn contained in all fish species could contribute adverse health effects to the local residents (hazard quotients higher than 1). Therefore, risk management measures must be promoted and implemented in all study areas to reduce potential risks to local Vietnamese residents. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Hybrid Process Fidelity Assessment in a Home-based Randomized Clinical Trial

PubMed Central

WILDE, MARY H.; LIEBEL, DIANNE; FAIRBANKS, EILEEN; WILSON, PAULA; LASH, MARGARET; SHAH, SHIVANI; McDONALD, MARGARET V.; BRASCH, JUDITH; ZHANG, FENG; SCHEID, EILEEN; McMAHON, JAMES M.

2016-01-01

A process fidelity assessment was conducted as a nested study within a home-based randomized clinical trial teaching self-management to 101 long-term indwelling urinary catheter users in the treatment group. Our hybrid model combined external assessments (outside observations and tape recordings) with internal evaluation methods (through study nurse forms and notes) for a comprehensive process fidelity assessment. Barriers, patient-related issues, and nurse perspectives were identified demonstrating the complexity in home care intervention research. The complementary and synergistic approaches provided in depth information about the context of the delivery and the impact of the intervention on study outcomes. PMID:25894688

State-of-the-art assessment of electric and hybrid vehicles

NASA Technical Reports Server (NTRS)

1978-01-01

Data are presented that were obtained from the electric and hybrid vehicles tested, information collected from users of electric vehicles, and data and information on electric and hybrid vehicles obtained on a worldwide basis from manufacturers and available literature. The data given include: (1) information and data base (electric and hybrid vehicle systems descriptions, sources of vehicle data and information, and sources of component data); (2) electric vehicles (theoretical background, electric vehicle track tests, user experience, literature data, and summary of electric vehicle status); (3) electric vehicle components (tires, differentials, transmissions, traction motors, controllers, batteries, battery chargers, and component summary); and (4) hybrid vehicles (types of hybrid vehicles, operating modes, hybrid vehicles components, and hybrid vehicles performance characteristics).

Biological assessments of Appalachian streams based on predictive models for fish, macroinvertebrate, and diatom assemblages

USGS Publications Warehouse

Carlisle, D.M.; Hawkins, C.P.; Meador, M.R.; Potapova, M.; Falcone, J.

2008-01-01

We developed empirical models for fish, macroinvertebrate, and diatom assemblages to assess the biological condition of 268 streams sampled from 1993 to 2002 in 7 major river basins in the Appalachian region of the USA. These models estimate the expected taxonomic composition at each site based on observed variation in taxonomic composition at reference sites. The index, O/E, is the ratio of the number of predicted taxa that were observed (O) to that expected (E) to occur at a site and is a measure of taxonomic completeness. We compared how O/E for each assemblage varied among major landuse settings and whether impaired assemblages were associated with particular physicochemical conditions. We also examined concordance among assemblages in their response to stress. Biological, chemical, and physical data were collected following consistent protocols. We used land-cover criteria, published data, and topographic maps to classify sites by major landuse setting. Fish, macroinvertebrate, and diatom assemblages had been sampled at 73, 108, and 52, respectively, of the least disturbed sites used to establish reference conditions. The models accounted for a substantial portion of the natural variation in taxonomic composition across sites that was associated with biogeographic, climatic, and basin-scale factors and generally were unbiased across the range of environmental gradients observed in the region. Assessments at nonreference sites showed that impairment of fish and macroinvertebrate assemblages was most strongly associated with agriculture and urban land uses, whereas impairment of diatom assemblages was most strongly associated with mining in the basin. Concordance in assessments among assemblages was not strong. Assessments based on 2 assemblages differed in 28 to 57% of cases, and assessments were never concordant for cases where all 3 assemblages were sampled. Furthermore, only 1/2 of these cases would have been assessed as ecologically impaired had only 1

quenched-smFISH: Counting small RNA in Pathogenic Bacteria

NASA Astrophysics Data System (ADS)

Shepherd, Douglas; Li, Nan; Micheva-Viteva, Sofiya; Munsky, Brian; Hong-Geller, Elizabeth; Werner, James

2014-03-01

Here, we present a modification to single-molecule fluorescence in situ hybridization, quenched smFISH (q-smFISH), that enables quantitative detection and analysis of small RNA (sRNA) expressed in bacteria. We show that short nucleic acid targets can be detected when the background of unbound singly dye-labeled DNA oligomers is reduced through hybridization with a set of complementary DNA oligomers labeled with a fluorescence quencher. Exploiting an automated, multi-color wide-field microscope and GPU-accelerated data analysis package, we analyzed the statistics of sRNA expression in thousands of individual Yersinia pseudotuberculosis and Yersinia pestis bacteria before and during a simulated infection. Before infection, we find only a small fraction of either bacteria express the small RNAs YSR35 or YSP8. The copy numbers of these RNA are increased during simulated infection, suggesting a role in pathogenesis. The ability to directly quantify expression level changes of sRNA in single cells as a function of external stimuli provides key information on the role of sRNA in bacterial regulatory networks.

Public health assessment of dioxin-contaminated fish at former US airbase, Bien Hoa, Vietnam.

PubMed

Durant, James T; Boivin, Thomas G; Pohl, Hana R; Sinks, Thomas H

2015-01-01

Ponds at the former US airbase at Ben Hoa, Vietnam are contaminated with Agent Orange. The ponds had been used for aquaculture, and in all likelihood, fish from those ponds have been sold to the public. We assessed human exposure to 2,3,7,8-tetrachloro-dibenzo-dioxin (2,3,7,8-TCDD) in fish samples from the ponds. For on-base tilapia, muscle concentrations 2,3,7,8-TCDD ranged from 1.4 to 32.7 pg/g. Fat concentrations ranged from 73.3 to 3990 pg/g. Estimated human exposure doses exceed international guidelines and exceed 2,3,7,8-TCDD's lowest adverse effect levels. The Bien Hoa fishponds are a completed human pathway for TCDD exposure.

Assessing effects of water abstraction on fish assemblages in Mediterranean streams

USGS Publications Warehouse

Benejam, Lluis; Angermeier, Paul L.; Munne, Antoni; García-Berthou, Emili

2010-01-01

1. Water abstraction strongly affects streams in arid and semiarid ecosystems, particularly where there is a Mediterranean climate. Excessive abstraction reduces the availability of water for human uses downstream and impairs the capacity of streams to support native biota. 2. We investigated the flow regime and related variables in six river basins of the Iberian Peninsula and show that they have been strongly altered, with declining flows (autoregressive models) and groundwater levels during the 20th century. These streams had lower flows and more frequent droughts than predicted by the official hydrological model used in this region. Three of these rivers were sometimes dry, whereas there were predicted by the model to be permanently flowing. Meanwhile, there has been no decrease in annual precipitation. 3. We also investigated the fish assemblage of a stream in one of these river basins (Tordera) for 6 years and show that sites more affected by water abstraction display significant differences in four fish metrics (catch per unit effort, number of benthic species, number of intolerant species and proportional abundance of intolerant individuals) commonly used to assess the biotic condition of streams. 4. We discuss the utility of these metrics in assessing impacts of water abstraction and point out the need for detailed characterisation of the natural flow regime (and hence drought events) prior to the application of biotic indices in streams severely affected by water abstraction. In particular, in cases of artificially dry streams, it is more appropriate for regulatory agencies to assign index scores that reflect biotic degradation than to assign ‘missing’ scores, as is presently customary in assessments of Iberian streams.

Optimizing fish sampling for fish - mercury bioaccumulation factors

USGS Publications Warehouse

Scudder Eikenberry, Barbara C.; Riva-Murray, Karen; Knightes, Christopher D.; Journey, Celeste A.; Chasar, Lia C.; Brigham, Mark E.; Bradley, Paul M.

2015-01-01

Fish Bioaccumulation Factors (BAFs; ratios of mercury (Hg) in fish (Hgfish) and water (Hgwater)) are used to develop Total Maximum Daily Load and water quality criteria for Hg-impaired waters. Both applications require representative Hgfish estimates and, thus, are sensitive to sampling and data-treatment methods. Data collected by fixed protocol from 11 streams in 5 states distributed across the US were used to assess the effects of Hgfish normalization/standardization methods and fish sample numbers on BAF estimates. Fish length, followed by weight, was most correlated to adult top-predator Hgfish. Site-specific BAFs based on length-normalized and standardized Hgfish estimates demonstrated up to 50% less variability than those based on non-normalized Hgfish. Permutation analysis indicated that length-normalized and standardized Hgfish estimates based on at least 8 trout or 5 bass resulted in mean Hgfish coefficients of variation less than 20%. These results are intended to support regulatory mercury monitoring and load-reduction program improvements.

Environmental assessment, K Pool fish rearing, Hanford Site, Richland, Washington

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1996-12-01

The US Department of Energy (DOE) has a need to respond to a request to lease facilities at the Hanford Site 100-KE and 100-KW filter plant pools (K Pools) for fish rearing activities. These fish rearing activities would be: (1) business ventures with public and private funds and (2) long-term enhancement and supplementation programs for game fish populations in the Columbia River Basin. The proposed action is to enter into a use permit or lease agreement with the YIN or other parties who would rear fish in the 100-K Area Pools. The proposed action would include necessary piping, pump, andmore » electrical upgrades of the facility; cleaning and preparation of the pools; water withdrawal from the Columbia River, and any necessary water or wastewater treatment; and introduction, rearing and release of fish. Future commercial operations may be included.« less

Hybrid Vehicle Technology Constraints and Application Assessment Study : Volume 3. Sections 5 through 9.

DOT National Transportation Integrated Search

1977-11-01

This four-volume report presents analyses and assessments of both heat engine/battery- and heat engine/flywheel-powered hybrid vehicles to determine if they could contribute to near-term (1980-1990) reductions in transportation energy consumption und...

Hybrid Vehicle Technology Constraints and Application Assessment Study : Volume 2. Sections 1 through 4.

DOT National Transportation Integrated Search

1977-01-01

This four-volume report presents analyses and assessments of both heat engine/battery- and heat engine/flywheel-powered hybrid vehicles to determine if they could contribute to near-term (1980-1990) reductions in transportation energy consumption und...

Dissemination of streptococcal pyrogenic exotoxin G (spegg) with an IS-like element in fish isolates of Streptococcus dysgalactiae.

PubMed

Abdelsalam, Mohamed; Chen, Shih-Chu; Yoshida, Terutoyo

2010-08-01

The Lancefield group C alpha-hemolytic Streptococcus dysgalactiae ssp. dysgalactiae (GCSD) causes systemic granulomatous inflammatory disease and high mortality rates in infected fish. Superantigen and streptolysin S genes are the most important virulence factors contributing to an invasive streptococcal infection. PCR amplification revealed that all strains isolated from moribund fish harbored the streptolysin S structural gene (sagA). GCSD fish isolates were PCR negative for emm, speA, speB, speC, speM, smeZ, and ssa. However, the size of the streptococcal pyrogenic exotoxin G (spegg) locus, a superantigen, in positive S. dysgalactiae fish and pig strains was variable. The ORF of the spegg locus of 26 GCSD fish strains and one GCSD pig strain was inserted with IS981SC. Interestingly, the ORF of the spegg locus of two fish strains of GCSD collected in Malaysia was inserted with an IS981SC-IS1161 hybrid IS element. The hybrid IS element was found in all of the GCSD fish isolates and one GCSD pig through PCR screening. Although no insertion sequence (IS) was detected in the spegg locus of S. dysgalactiae ssp. equisimilis (GCSE) strains, a five-nucleotide deletion mutation was detected in the ORF of the spegg locus of one GCSE strain at the supposed site of IS981SC insertion, resulting in a frameshift mutation.

Changes in the fish fauna of the Kissimmee River basin, peninsular Florida: Nonnative additions

USGS Publications Warehouse

Nico, L.G.

2005-01-01

Recent decades have seen substantial changes in fish assemblages in rivers of peninsular Florida. The most striking change has involved the addition of nonnative fishes, including taxa from Asia, Africa, and Central and South America. I review recent and historical records of fishes occurring in the Kissimmee River basin (7,800 km2), a low-gradient drainage with 47 extant native fishes (one possibly the result of an early transplant), at least 7 foreign fishes (most of which are widely established), and a stocked hybrid. Kissimmee assemblages include fewer marine fishes than the nearby Peace and Caloosahatchee rivers, and fewer introduced foreign fishes than south Florida canals. Fish assemblages of the Kissimmee and other subtropical Florida rivers are dynamic, due to new introductions, range expansions of nonnative fishes already present, and periodic declines in nonnative fish populations during occasional harsh winters. The addition, dispersal, and abundance of nonnative fishes in the basin is linked to many factors, including habitat disturbance, a subtropical climate, and the fact that the basin is centrally located in a region where drainage boundaries are blurred and introductions of foreign fishes commonplace. The first appearance of foreign fishes in the basin coincided with the complete channelization of the Kissimmee River in the 1970s. Although not a causal factor, artificial waterways connecting the upper lakes and channelization of the Kissimmee River have facilitated dispersal. With one possible exception, there have been no basin-wide losses of native fishes. When assessing change in peninsular Florida waters, extinction or extirpation of fishes appears to be a poor measure of impact. No endemic species are known from peninsular Florida (although some endemic subspecies have been noted). Most native freshwater fishes are themselves descended from recent invaders that reached the peninsula from the main continent. These invasions likely were

Karyotype variability in tropical maize sister inbred lines and hybrids compared with KYS standard line

PubMed Central

Mondin, Mateus; Santos-Serejo, Janay A.; Bertäo, Mônica R.; Laborda, Prianda; Pizzaia, Daniel; Aguiar-Perecin, Margarida L. R.

2014-01-01

Maize karyotype variability has been extensively investigated. The identification of maize somatic and pachytene chromosomes has improved with the development of fluorescence in situ hybridization (FISH) using tandemly repeated DNA sequences as probes. We identified the somatic chromosomes of sister inbred lines that were derived from a tropical flint maize population (Jac Duro [JD]), and hybrids between them, using FISH probes for the 180-bp knob repeat, centromeric satellite (CentC), centromeric satellite 4 (Cent4), subtelomeric clone 4-12-1, 5S ribosomal DNA and nucleolus organizing region DNA sequences. The observations were integrated with data based on C-banded mitotic metaphases and conventional analysis of pachytene chromosomes. Heterochromatic knobs visible at pachynema were coincident with C-bands and 180-bp FISH signals on somatic chromosomes, and most of them were large. Variation in the presence of some knobs was observed among lines. Small 180-bp knob signals were invariant on the short arms of chromosomes 1, 6, and 9. The subtelomeric 4-12-1 signal was also invariant and useful for identifying some chromosomes. The centromere location of chromosomes 2 and 4 differed from previous reports on standard maize lines. Somatic chromosomes of a JD line and the commonly used KYS line were compared by FISH in a hybrid of these lines. The pairing behavior of chromosomes 2 and 4 at pachytene stage in this hybrid was investigated using FISH with chromosome-specific probes. The homologues were fully synapsed, including the 5S rDNA and CentC sites on chromosome 2, and Cent4 and subtelomeric 4-12-1 sites on chromosome 4. This suggests that homologous chromosomes could pair through differential degrees of chromatin packaging in homologous arms differing in size. The results contribute to current knowledge of maize global diversity and also raise questions concerning the meiotic pairing of homologous chromosomes possibly differing in their amounts of repetitive DNA

A comparative assessment of contaminants in fish from four resacas of the Texas, USA-Tamaulipas, Mexico border region

USGS Publications Warehouse

Mora, Miguel A.; Papoulias, Diana M.; Nava, Ismael; Buckler, Denny R.

2001-01-01

A recent survey of contaminant information for the Lower Rio Grande Valley (LRGV), Texas, has shown that little is known about contaminants and their impacts on biota of resacas (oxbows) along the US-Mexico border. In 1996, fish were collected from four resacas in the Texas- Tamaulipas border region to assess contaminant loadings and their impacts on fish and birds. Tissue residue concentrations in fish were analyzed and also compared to two histopathological bioindicators of unhealthy environmental conditions. Of the organochlorine insecticides measured, DDE was the most common and was present at relatively high concentrations (10 ??g/g w/w) at some sites. DDE concentrations were nearly 20 times greater in fish from resacas in Texas than from resacas in Tamaulipas, although the limited sample sizes obtained precluded statistical comparisons. DDE concentrations in fish from the two Texas resacas were also greater than those reported in fish from nearby areas during the 1980s and 1990s. Most trace element concentrations were similar among resacas from Texas and Tamaulipas. Arsenic, however, was two to six times greater in fish from a downtown resaca in Matamoros than in fish from other resacas in Tamaulipas and Texas. The bioindicators, pigment accumulation, and macrophage aggregates (MAs), in general, reflected the contamination indicated by the tissue residues for each site. Overall, it appears that some resacas of the US-Mexico border region are contaminant sinks and could pose potential health or reproductive problems for fish and wildlife, and humans that consume fish from those sites. Published by Elsevier Science Ltd.

Concentrations of heavy metals in marine wild fishes captured from the southern sea of Korea and associated health risk assessments

NASA Astrophysics Data System (ADS)

Hwang, Dong-Woon; Kim, Seong-Soo; Kim, Seong-Gil; Kim, Dong-Sun; Kim, Tae-Hoon

2017-12-01

Concentrations of heavy metals (As, Cd, Cr, Cu, Hg, Pb, and Zn) were determined in edible parts (muscle) of 34 marine wild fish caught from the southern sea of Korea in 2007 and 2008 in order to understand the accumulation pattern of heavy metals in wild fish and to assess the potential health risk posed by fish consumption. The highest concentrations in the muscle of 17 pelagic and 17 demersal fishes were Zn and As, respectively, while the lowest concentration in both fishes was Cd. The mean concentrations of all metals except As in wild fish were much lower than the regulatory limits for fish and fishery products applied in a number of countries. Unlike other metals, As concentration in wild fish of this study region was relatively higher than that found in other country. Estimated daily intake (EDI) of the metals was in the range of 0.05% to 22.5% of the provisional maximum tolerable daily intakes (PMTDI). Similarly, the target hazard quotient (THQ) was below 1.0 for each metal. These results imply that the consumption of the investigated wild fish do not cause significant adverse health effects.

Hybrid Reynolds-Averaged/Large Eddy Simulation of a Cavity Flameholder; Assessment of Modeling Sensitivities

NASA Technical Reports Server (NTRS)

Baurle, R. A.

2015-01-01

Steady-state and scale-resolving simulations have been performed for flow in and around a model scramjet combustor flameholder. The cases simulated corresponded to those used to examine this flowfield experimentally using particle image velocimetry. A variety of turbulence models were used for the steady-state Reynolds-averaged simulations which included both linear and non-linear eddy viscosity models. The scale-resolving simulations used a hybrid Reynolds-averaged / large eddy simulation strategy that is designed to be a large eddy simulation everywhere except in the inner portion (log layer and below) of the boundary layer. Hence, this formulation can be regarded as a wall-modeled large eddy simulation. This effort was undertaken to formally assess the performance of the hybrid Reynolds-averaged / large eddy simulation modeling approach in a flowfield of interest to the scramjet research community. The numerical errors were quantified for both the steady-state and scale-resolving simulations prior to making any claims of predictive accuracy relative to the measurements. The steady-state Reynolds-averaged results showed a high degree of variability when comparing the predictions obtained from each turbulence model, with the non-linear eddy viscosity model (an explicit algebraic stress model) providing the most accurate prediction of the measured values. The hybrid Reynolds-averaged/large eddy simulation results were carefully scrutinized to ensure that even the coarsest grid had an acceptable level of resolution for large eddy simulation, and that the time-averaged statistics were acceptably accurate. The autocorrelation and its Fourier transform were the primary tools used for this assessment. The statistics extracted from the hybrid simulation strategy proved to be more accurate than the Reynolds-averaged results obtained using the linear eddy viscosity models. However, there was no predictive improvement noted over the results obtained from the explicit

A novel approach to assessing environmental disturbance based on habitat selection by zebra fish as a model organism.

PubMed

Araújo, Cristiano V M; Griffith, Daniel M; Vera-Vera, Victoria; Jentzsch, Paul Vargas; Cervera, Laura; Nieto-Ariza, Beatriz; Salvatierra, David; Erazo, Santiago; Jaramillo, Rusbel; Ramos, Luis A; Moreira-Santos, Matilde; Ribeiro, Rui

2018-04-01

Aquatic ecotoxicity assays used to assess ecological risk assume that organisms living in a contaminated habitat are forcedly exposed to the contamination. This assumption neglects the ability of organisms to detect and avoid contamination by moving towards less disturbed habitats, as long as connectivity exists. In fluvial systems, many environmental parameters vary spatially and thus condition organisms' habitat selection. We assessed the preference of zebra fish (Danio rerio) when exposed to water samples from two western Ecuadorian rivers with apparently distinct disturbance levels: Pescadillo River (highly disturbed) and Oro River (moderately disturbed). Using a non-forced exposure system in which water samples from each river were arranged according to their spatial sequence in the field and connected to allow individuals to move freely among samples, we assayed habitat selection by D. rerio to assess environmental disturbance in the two rivers. Fish exposed to Pescadillo River samples preferred downstream samples near the confluence zone with the Oro River. Fish exposed to Oro River samples preferred upstream waters. When exposed to samples from both rivers simultaneously, fish exhibited the same pattern of habitat selection by preferring the Oro River samples. Given that the rivers are connected, preference for the Oro River enabled us to predict a depression in fish populations in the Pescadillo River. Although these findings indicate higher disturbance levels in the Pescadillo River, none of the physical-chemical variables measured was significantly correlated with the preference pattern towards the Oro River. Non-linear spatial patterns of habitat preference suggest that other environmental parameters like urban or agricultural contaminants play an important role in the model organism's habitat selection in these rivers. The non-forced exposure system represents a habitat selection-based approach that can serve as a valuable tool to unravel the factors

Assessing three fish species ecological status in Colorado River, Grand Canyon based on physical habitat and population models.

PubMed

Yao, Weiwei; Chen, Yuansheng

2018-04-01

Colorado River is a unique ecosystem and provides important ecological services such as habitat for fish species as well as water power energy supplies. River management for this ecosystem requires assessment and decision support tools for fish which involves protecting, restoring as well as forecasting of future conditions. In this paper, a habitat and population model was developed and used to determine the levels of fish habitat suitability and population density in Colorado River between Lees Ferry and Lake Mead. The short term target fish populations are also predicted based on native fish recovery strategy. This model has been developed by combining hydrodynamics, heat transfer and sediment transport models with a habitat suitability index model and then coupling with habitat model into life stage population model. The fish were divided into four life stages according to the fish length. Three most abundant and typical native and non-native fish were selected as target species, which are rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta) and flannelmouth sucker (Catostomus latipinnis). Flow velocity, water depth, water temperature and substrates were used as the suitability indicators in habitat model and overall suitability index (OSI) as well as weight usable area (WUA) was used as an indicator in population model. A comparison was made between simulated fish population alteration and surveyed fish number fluctuation during 2000 to 2009. The application of this habitat and population model indicates that this model can be accurate present habitat situation and targets fish population dynamics of in the study areas. The analysis also indicates the flannelmouth sucker population will steadily increase while the rainbow trout will decrease based on the native fish recovery scheme. Copyright © 2018. Published by Elsevier Inc.

The development of a preliminary rock reef fish multimetric index for assessing thermal and urban impacts in a tropical bay.

PubMed

Teixeira-Neves, Tatiana Pires; Neves, Leonardo Mitrano; Araújo, Francisco Gerson

2016-08-15

We developed a multimetric index for assessing ecological conditions in rocky reefs areas to evaluate thermal and urban influences on fish community. Eight metrics were selected to assess thermal influence: (1) total number of species; (2) number of water column species; (3) number of transient species; (4) density of individuals with low resilience; (5) density of omnivores; (6) density of carnivores; (7) number of cryptic species; (8) density of herbivores. For urban influence, six metrics were selected: (1) total density; (2) ratio between the number of rare species and the total number of species; (3) density of individuals with heavy fishing pressure; (4) number of resident species; (5) number of cryptic species; (6) density of herbivores. This preliminary index succeed in discriminating control/impacted sites and proved to be an important tool to assess impacts that alter fish community and have potential to be used in tropical rock reef coastal areas. Copyright © 2016 Elsevier Ltd. All rights reserved.

Use of fluorescence in situ hybridization as a tool for introgression analysis and chromosome identification in coffee (Coffea arabica L.).

PubMed

Herrera, Juan Carlos; D'Hont, Angelique; Lashermes, Philippe

2007-07-01

Fluorescence in situ hybridization (FISH) was used to study the presence of alien chromatin in interspecific hybrids and one introgressed line (S.288) derived from crosses between the cultivated species Coffea arabica and the diploid relatives C. canephora and C. liberica. In situ hybridization using genomic DNA from C. canephora and C. arabica as probes showed elevated cross hybridization along the hybrid genome, confirming the weak differentiation between parental genomes. According to our genomic in situ hybridization (GISH) data, the observed genomic resemblance between the modern C. canephora genome (C) and the C. canephora-derived subgenome of C. arabica (Ca) appears rather considerable. Poor discrimination between C and Ca chromosomes supports the idea of low structural modifications of both genomes since the C. arabica speciation, at least in the frequency and distribution of repetitive sequences. GISH was also used to identify alien chromatin segments on chromosome spreads of a C. liberica-introgressed line of C. arabica. Further, use of GISH together with BAC-FISH analysis gave us additional valuable information about the physical localization of the C. liberica fragments carrying the SH3 factor involved in resistance to the coffee leaf rust. Overall, our results illustrate that FISH analysis is a complementary tool for molecular cytogenetic studies in coffee, providing rapid localization of either specific chromosomes or alien chromatin in introgressed genotypes derived from diploid species displaying substantial genomic differentiation from C. arabica.

In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea

PubMed Central

2013-01-01

Background The freshwater planarian Schmidtea mediterranea has emerged as a powerful model for studies of regenerative, stem cell, and germ cell biology. Whole-mount in situ hybridization (WISH) and whole-mount fluorescent in situ hybridization (FISH) are critical methods for determining gene expression patterns in planarians. While expression patterns for a number of genes have been elucidated using established protocols, determining the expression patterns for particularly low-abundance transcripts remains a challenge. Results We show here that a short bleaching step in formamide dramatically enhances signal intensity of WISH and FISH. To further improve signal sensitivity we optimized blocking conditions for multiple anti-hapten antibodies, developed a copper sulfate quenching step that virtually eliminates autofluorescence, and enhanced signal intensity through iterative rounds of tyramide signal amplification. For FISH on regenerating planarians, we employed a heat-induced antigen retrieval step that provides a better balance between permeabilization of mature tissues and preservation of regenerating tissues. We also show that azide most effectively quenches peroxidase activity between rounds of development for multicolor FISH experiments. Finally, we apply these modifications to elucidate the expression patterns of a few low-abundance transcripts. Conclusion The modifications we present here provide significant improvements in signal intensity and signal sensitivity for WISH and FISH in planarians. Additionally, these modifications might be of widespread utility for whole-mount FISH in other model organisms. PMID:23497040

Fishing drives declines in fish parasite diversity and has variable effects on parasite abundance.

PubMed

Wood, Chelsea L; Sandin, Stuart A; Zgliczynski, Brian; Guerra, Ana Sofía; Micheli, Fiorenza

2014-07-01

Despite the ubiquity and ecological importance of parasites, relatively few studies have assessed their response to anthropogenic environmental change. Heuristic models have predicted both increases and decreases in parasite abundance in response to human disturbance, with empirical support for both. However, most studies focus on one or a few selected parasite species. Here, we assess the abundance of parasites of seven species of coral reef fishes collected from three fished and three unfished islands of the Line Islands archipelago in the central equatorial Pacific. Because we chose fish hosts that spanned different trophic levels, taxonomic groups, and body sizes, we were able to compare parasite responses across a broad cross section of the total parasite community in the presence and absence of fishing, a major human impact on marine ecosystems. We found that overall parasite species richness was substantially depressed on fished islands, but that the response of parasite abundance varied among parasite taxa: directly transmitted parasites were significantly more abundant on fished than on unfished islands, while the reverse was true for trophically transmitted parasites. This probably arises because trophically transmitted parasites require multiple host species, some of which are the top predators most sensitive to fishing impacts. The increase in directly transmitted parasites appeared to be due to fishing-driven compensatory increases in the abundance of their hosts. Together, these results provide support for the predictions of both heuristic models, and indicate that the direction of fishing's impact on parasite abundance is mediated by parasite traits, notably parasite transmission strategies.

Effects of salinity on gastric emptying time in hybrid grouper, Epinephelus fuscoguttatus x E. lanceolattus juveniles

NASA Astrophysics Data System (ADS)

Noor, Noorashikin Md.; Das, Simon Kumar; Cob, Zaidi Che; Ghaffar, Mazlan Abd.

2018-04-01

The newly developed hybrid grouper: tiger grouper (Epinephelus fuscoguttatus) × giant grouper (Epinephelus fuscoguttatus) (TG×GG), has a high resistance towards different environmental condition (eg. in euryhaline environment) due to its genetic improvement. This study aims to investigate the effects of different salinities (10, 15, 20, 25 and 30 ppt) on the gastric emptying time (GET) of the TG×GG hybrid grouper juveniles. The fish were fed with commercial pellet over a 30 days experimental period under controlled laboratory conditions. The GET was determined by X-radiographic method, using barium sulfate (BaSO4) as an inert food marker. The X-radiography images showed that the shortest GET (12 h) was observed in the 15 ppt group, whereas the longest GET (18 h) in 30 ppt group. The results suggests to culture TG×GG hybrid grouper juveniles in 15 ppt with commercial pellet diet as this salinity proliferates faster digestion process which may contribute faster growth rate of this important fish species. Overall, these findings would be useful for the betterment of TG×GG hybrid grouper aquaculture which will eventually boost up the production of this newly developed hybrid grouper species.

Bioavailability of pharmaceuticals in waters close to wastewater treatment plants: use of fish bile for exposure assessment.

PubMed

Lahti, Marja; Brozinski, Jenny-Maria; Segner, Helmut; Kronberg, Leif; Oikari, Aimo

2012-08-01

Pharmaceuticals are ubiquitous in surface waters as a consequence of discharges from municipal wastewater treatment plants. However, few studies have assessed the bioavailability of pharmaceuticals to fish in natural waters. In the present study, passive samplers and rainbow trout were experimentally deployed next to three municipal wastewater treatment plants in Finland to evaluate the degree of animal exposure. Pharmaceuticals from several therapeutic classes (in total 15) were analyzed by liquid chromatography-tandem mass spectrometry in extracts of passive samplers and in bile and blood plasma of rainbow trout held at polluted sites for 10 d. Each approach indicated the highest exposure near wastewater treatment plant A and the lowest near that of plant C. Diclofenac, naproxen, and ibuprofen were found in rainbow trout, and their concentrations in bile were 10 to 400 times higher than in plasma. The phase I metabolite hydroxydiclofenac was also detected in bile. Hence, bile proved to be an excellent sample matrix for the exposure assessment of fish. Most of the monitored pharmaceuticals were found in passive samplers, implying that they may overestimate the actual exposure of fish in receiving waters. Two biomarkers, hepatic vitellogenin and cytochrome P4501A, did not reveal clear effects on fish, although a small induction of vitellogenin mRNA was observed in trout caged near wastewater treatment plants B and C. Copyright © 2012 SETAC.

The fishes of George Washington Carver National Monument, Missouri, 2003

USGS Publications Warehouse

Justus, B.G.; Petersen, James C.

2005-01-01

Fish were collected at six sites at George Washington Carver National Monument by seining and electrofishing during a base-flow period on July 17-18, 2003. Approximately 700 fish were collected and identified at the six sampling sites. Those individuals represented 17 species (and 1 hybrid) and 13 genera. The number of species collected at the five stream sites ranged from 9 to 12; a hybrid sunfish and 4 species were collected from a pond. Fish collected at stream sites were typical of small headwater streams and no species collected in this study are federally-listed threatened or endangered species. The three most common species were the southern redbelly dace, central stoneroller, and green sunfish. Some differences existed between the assemblages (groups of species) collected in 2003 and in the previous inventories. Four of the 17 fish species collected in this inventory previously had not been collected at the monument. However, 11 species collected in one or more of the previous inventories were not collected in this effort. There is no indication that a change in environmental conditions is responsible for the absence of these species; more likely reasons are seasonal variability, extirpation, low population density, and misidentification. Four species collected at George Washington Carver National Monument may be of special interest to National Park Service managers and others. The cardinal shiner and stippled darter are endemic to the Ozark Plateaus. The Arkansas darter is considered a species of conservation concern by the State of Missouri. The grass carp is an introduced species.

Improving Hybrid III injury assessment in steering wheel rim to chest impacts using responses from finite element Hybrid III and human body model.

PubMed

Holmqvist, Kristian; Davidsson, Johan; Mendoza-Vazquez, Manuel; Rundberget, Peter; Svensson, Mats Y; Thorn, Stefan; Törnvall, Fredrik

2014-01-01

The main aim of this study was to improve the quality of injury risk assessments in steering wheel rim to chest impacts when using the Hybrid III crash test dummy in frontal heavy goods vehicle (HGV) collision tests. Correction factors for chest injury criteria were calculated as the model chest injury parameter ratios between finite element (FE) Hybrid III, evaluated in relevant load cases, and the Total Human Model for Safety (THUMS). This is proposed to be used to compensate Hybrid III measurements in crash tests where steering wheel rim to chest impacts occur. The study was conducted in an FE environment using an FE-Hybrid III model and the THUMS. Two impactor shapes were used, a circular hub and a long, thin horizontal bar. Chest impacts at velocities ranging from 3.0 to 6.0 m/s were simulated at 3 impact height levels. A ratio between FE-Hybrid III and THUMS chest injury parameters, maximum chest compression C max, and maximum viscous criterion VC max, were calculated for the different chest impact conditions to form a set of correction factors. The definition of the correction factor is based on the assumption that the response from a circular hub impact to the middle of the chest is well characterized and that injury risk measures are independent of impact height. The current limits for these chest injury criteria were used as a basis to develop correction factors that compensate for the limitations in biofidelity of the Hybrid III in steering wheel rim to chest impacts. The hub and bar impactors produced considerably higher C max and VC max responses in the THUMS compared to the FE-Hybrid III. The correction factor for the responses of the FE-Hybrid III showed that the criteria responses for the bar impactor were consistently overestimated. Ratios based on Hybrid III and THUMS responses provided correction factors for the Hybrid III responses ranging from 0.84 to 0.93. These factors can be used to estimate C max and VC max values when the Hybrid III is

A retrospective on hydroacoustic assessment of fish passage in Alaskan rivers

NASA Astrophysics Data System (ADS)

Burwen, Debby; Fleischman, Steve; Maxwell, Suzanne; Pfisterer, Carl

2005-04-01

The Alaska Department of Fish and Game (ADFG) has enumerated fish stocks in rivers for over 30 years using a variety of acoustic technologies including single-, dual-, and split-beam sonar. Most recently, ADFG has evaluated a relatively new sonar technology at several sites in Alaska to determine its applicability to counting migrating fish in rivers. The new system, called a Dual frequency IDentification SONar (DIDSON), is a high-definition imaging sonar designed and manufactured by the University of Washington's Applied Physics Lab for military applications such as diver detection and underwater mine identification. Results from experiments conducted in 2002-2004 indicate that DIDSON provides significant improvements in our ability to detect, track, and determine the direction of travel of migrating fish in rivers. One of the most powerful uses of the DIDSON has been to combine its camera-like images of fish swimming behavior with corresponding split-beam data. These linked datasets have allowed us to evaluate the effects of fish orientation and swimming behavior on echo shape parameters that have proven useful in the classification of certain fish species.

Genetic investigation of natural hybridization between rainbow and coastal cutthroat trout in the copper River Delta, Alaska

USGS Publications Warehouse

Williams, I.; Reeves, G.H.; Graziano, S.L.; Nielsen, J.L.

2007-01-01

Molecular genetic methods were used to quantify natural hybridization between rainbow trout Oncorhynchus mykiss or steelhead (anadromous rainbow trout) and coastal cutthroat trout O. clarkii clarkii collected in the Copper River delta, Southeast Alaska. Eleven locations were sampled to determine the extent of hybridization and the distribution of hybrids. Four diagnostic nuclear microsatellite loci and four species-specific simple sequence repeat markers were used in combination with restriction fragment length polymorphism analyses of NADH dehydrogenase 5/6 (ND5/6) mitochondrial DNA (mtDNA) to investigate the genetic structure of trout from both species and identify putative interspecific hybrids. Hybrids were found in 7 of the 11 streams sampled in the Copper River delta, the extent of hybridization across all streams varying from 0% to 58%. Hybrid trout distribution appeared to be nonrandom, most individuals of mixed taxonomic ancestry being detected in streams containing rainbow trout rather than in streams containing coastal cutthroat trout. Genotypic disequilibrium was observed among microsatellite loci in populations with high levels of hybridization. We found no significant correlation between unique stream channel process groups and the number of hybrid fish sampled. Eighty-eight percent of fish identified as first-generation hybrids (F1) in two populations contained coastal cutthroat trout mtDNA, suggesting directionality in hybridization. However, dominance of coastal cutthroat trout mtDNA was not observed at a third location containing F1 hybrids, indicating that interspecific mating behavior varied among locations. Backcrossed individuals were found in drainages lacking F1 hybrids and in populations previously thought to contain a single species. The extent and distribution of backcrossed individuals suggested that at least some hybrids are reproductively viable and backcrossed hybrid offspring move throughout the system.

Comprehensive Genomic Profiling Identifies a Subset of Crizotinib-Responsive ALK-Rearranged Non-Small Cell Lung Cancer Not Detected by Fluorescence In Situ Hybridization

PubMed Central

Hensing, Thomas; Schrock, Alexa B.; Allen, Justin; Sanford, Eric; Gowen, Kyle; Kulkarni, Atul; He, Jie; Suh, James H.; Lipson, Doron; Elvin, Julia A.; Yelensky, Roman; Chalmers, Zachary; Chmielecki, Juliann; Peled, Nir; Klempner, Samuel J.; Firozvi, Kashif; Frampton, Garrett M.; Molina, Julian R.; Menon, Smitha; Brahmer, Julie R.; MacMahon, Heber; Nowak, Jan; Ou, Sai-Hong Ignatius; Zauderer, Marjorie; Ladanyi, Marc; Zakowski, Maureen; Fischbach, Neil; Ross, Jeffrey S.; Stephens, Phil J.; Miller, Vincent A.; Wakelee, Heather

2016-01-01

Introduction. For patients with non-small cell lung cancer (NSCLC) to benefit from ALK inhibitors, sensitive and specific detection of ALK genomic rearrangements is needed. ALK break-apart fluorescence in situ hybridization (FISH) is the U.S. Food and Drug Administration approved and standard-of-care diagnostic assay, but identification of ALK rearrangements by other methods reported in NSCLC cases that tested negative for ALK rearrangements by FISH suggests a significant false-negative rate. We report here a large series of NSCLC cases assayed by hybrid-capture-based comprehensive genomic profiling (CGP) in the course of clinical care. Materials and Methods. Hybrid-capture-based CGP using next-generation sequencing was performed in the course of clinical care of 1,070 patients with advanced lung cancer. Each tumor sample was evaluated for all classes of genomic alterations, including base-pair substitutions, insertions/deletions, copy number alterations and rearrangements, as well as fusions/rearrangements. Results. A total of 47 patients (4.4%) were found to harbor ALK rearrangements, of whom 41 had an EML4-ALK fusion, and 6 had other fusion partners, including 3 previously unreported rearrangement events: EIF2AK-ALK, PPM1B-ALK, and PRKAR1A-ALK. Of 41 patients harboring ALK rearrangements, 31 had prior FISH testing results available. Of these, 20 were ALK FISH positive, and 11 (35%) were ALK FISH negative. Of the latter 11 patients, 9 received crizotinib based on the CGP results, and 7 achieved a response with median duration of 17 months. Conclusion. Comprehensive genomic profiling detected canonical ALK rearrangements and ALK rearrangements with noncanonical fusion partners in a subset of patients with NSCLC with previously negative ALK FISH results. In this series, such patients had durable responses to ALK inhibitors, comparable to historical response rates for ALK FISH-positive cases. Implications for Practice: Comprehensive genomic profiling (CGP) that

Comprehensive Genomic Profiling Identifies a Subset of Crizotinib-Responsive ALK-Rearranged Non-Small Cell Lung Cancer Not Detected by Fluorescence In Situ Hybridization.

PubMed

Ali, Siraj M; Hensing, Thomas; Schrock, Alexa B; Allen, Justin; Sanford, Eric; Gowen, Kyle; Kulkarni, Atul; He, Jie; Suh, James H; Lipson, Doron; Elvin, Julia A; Yelensky, Roman; Chalmers, Zachary; Chmielecki, Juliann; Peled, Nir; Klempner, Samuel J; Firozvi, Kashif; Frampton, Garrett M; Molina, Julian R; Menon, Smitha; Brahmer, Julie R; MacMahon, Heber; Nowak, Jan; Ou, Sai-Hong Ignatius; Zauderer, Marjorie; Ladanyi, Marc; Zakowski, Maureen; Fischbach, Neil; Ross, Jeffrey S; Stephens, Phil J; Miller, Vincent A; Wakelee, Heather; Ganesan, Shridar; Salgia, Ravi

2016-06-01

For patients with non-small cell lung cancer (NSCLC) to benefit from ALK inhibitors, sensitive and specific detection of ALK genomic rearrangements is needed. ALK break-apart fluorescence in situ hybridization (FISH) is the U.S. Food and Drug Administration approved and standard-of-care diagnostic assay, but identification of ALK rearrangements by other methods reported in NSCLC cases that tested negative for ALK rearrangements by FISH suggests a significant false-negative rate. We report here a large series of NSCLC cases assayed by hybrid-capture-based comprehensive genomic profiling (CGP) in the course of clinical care. Hybrid-capture-based CGP using next-generation sequencing was performed in the course of clinical care of 1,070 patients with advanced lung cancer. Each tumor sample was evaluated for all classes of genomic alterations, including base-pair substitutions, insertions/deletions, copy number alterations and rearrangements, as well as fusions/rearrangements. A total of 47 patients (4.4%) were found to harbor ALK rearrangements, of whom 41 had an EML4-ALK fusion, and 6 had other fusion partners, including 3 previously unreported rearrangement events: EIF2AK-ALK, PPM1B-ALK, and PRKAR1A-ALK. Of 41 patients harboring ALK rearrangements, 31 had prior FISH testing results available. Of these, 20 were ALK FISH positive, and 11 (35%) were ALK FISH negative. Of the latter 11 patients, 9 received crizotinib based on the CGP results, and 7 achieved a response with median duration of 17 months. Comprehensive genomic profiling detected canonical ALK rearrangements and ALK rearrangements with noncanonical fusion partners in a subset of patients with NSCLC with previously negative ALK FISH results. In this series, such patients had durable responses to ALK inhibitors, comparable to historical response rates for ALK FISH-positive cases. Comprehensive genomic profiling (CGP) that includes hybrid capture and specific baiting of intron 19 of ALK is a highly sensitive

Phytoplankton IF-FISH: Species-specific labeling of cellular proteins by immunofluorescence (IF) with simultaneous species identification by fluorescence immunohybridization (FISH).

PubMed

Meek, Megan E; Van Dolah, Frances M

2016-05-01

Phytoplankton rarely occur as unialgal populations. Therefore, to study species-specific protein expression, indicative of physiological status in natural populations, methods are needed that will both assay for a protein of interest and identify the species expressing it. Here we describe a protocol for IF-FISH, a dual labeling procedure using immunofluorescence (IF) labeling of a protein of interest followed by fluorescence in situ hybridization (FISH) to identify the species expressing that protein. The protocol was developed to monitor expression of the cell cycle marker proliferating cell nuclear antigen (PCNA) in the red tide dinoflagellate, Karenia brevis, using a large subunit (LSU) rRNA probe to identify K. brevis in a mixed population of morphologically similar Karenia species. We present this protocol as proof of concept that IF-FISH can be successfully applied to phytoplankton cells. This method is widely applicable for the analysis of single-cell protein expression of any protein of interest within phytoplankton communities. Copyright © 2016 Elsevier B.V. All rights reserved.

A hybrid genetic linkage map of two ecologically and morphologically divergent Midas cichlid fishes (Amphilophus spp.) obtained by massively parallel DNA sequencing (ddRADSeq).

PubMed

Recknagel, Hans; Elmer, Kathryn R; Meyer, Axel

2013-01-01

Cichlid fishes are an excellent model system for studying speciation and the formation of adaptive radiations because of their tremendous species richness and astonishing phenotypic diversity. Most research has focused on African rift lake fishes, although Neotropical cichlid species display much variability as well. Almost one dozen species of the Midas cichlid species complex (Amphilophus spp.) have been described so far and have formed repeated adaptive radiations in several Nicaraguan crater lakes. Here we apply double-digest restriction-site associated DNA sequencing to obtain a high-density linkage map of an interspecific cross between the benthic Amphilophus astorquii and the limnetic Amphilophus zaliosus, which are sympatric species endemic to Crater Lake Apoyo, Nicaragua. A total of 755 RAD markers were genotyped in 343 F(2) hybrids. The map resolved 25 linkage groups and spans a total distance of 1427 cM with an average marker spacing distance of 1.95 cM, almost matching the total number of chromosomes (n = 24) in these species. Regions of segregation distortion were identified in five linkage groups. Based on the pedigree of parents to F(2) offspring, we calculated a genome-wide mutation rate of 6.6 × 10(-8) mutations per nucleotide per generation. This genetic map will facilitate the mapping of ecomorphologically relevant adaptive traits in the repeated phenotypes that evolved within the Midas cichlid lineage and, as the first linkage map of a Neotropical cichlid, facilitate comparative genomic analyses between African cichlids, Neotropical cichlids and other teleost fishes.

A Hybrid Genetic Linkage Map of Two Ecologically and Morphologically Divergent Midas Cichlid Fishes (Amphilophus spp.) Obtained by Massively Parallel DNA Sequencing (ddRADSeq)

PubMed Central

Recknagel, Hans; Elmer, Kathryn R.; Meyer, Axel

2013-01-01

Cichlid fishes are an excellent model system for studying speciation and the formation of adaptive radiations because of their tremendous species richness and astonishing phenotypic diversity. Most research has focused on African rift lake fishes, although Neotropical cichlid species display much variability as well. Almost one dozen species of the Midas cichlid species complex (Amphilophus spp.) have been described so far and have formed repeated adaptive radiations in several Nicaraguan crater lakes. Here we apply double-digest restriction-site associated DNA sequencing to obtain a high-density linkage map of an interspecific cross between the benthic Amphilophus astorquii and the limnetic Amphilophus zaliosus, which are sympatric species endemic to Crater Lake Apoyo, Nicaragua. A total of 755 RAD markers were genotyped in 343 F2 hybrids. The map resolved 25 linkage groups and spans a total distance of 1427 cM with an average marker spacing distance of 1.95 cM, almost matching the total number of chromosomes (n = 24) in these species. Regions of segregation distortion were identified in five linkage groups. Based on the pedigree of parents to F2 offspring, we calculated a genome-wide mutation rate of 6.6 × 10−8 mutations per nucleotide per generation. This genetic map will facilitate the mapping of ecomorphologically relevant adaptive traits in the repeated phenotypes that evolved within the Midas cichlid lineage and, as the first linkage map of a Neotropical cichlid, facilitate comparative genomic analyses between African cichlids, Neotropical cichlids and other teleost fishes. PMID:23316439

Assessing Potential Conservation and Restoration Areas of Freshwater Fish Fauna in the Indian River Basins.

PubMed

Bhatt, Jay P; Manish, Kumar; Mehta, Rajender; Pandit, Maharaj K

2016-05-01

Conservation efforts globally are skewed toward terrestrial ecosystems. To date, conservation of aquatic ecosystems, in particular fish fauna, is largely neglected. We provide a country-wide assessment of Indian river ecosystems in order to identify and prioritize areas for protection and restoration of freshwater fish fauna. Using various biodiversity and anthropogenic attributes, coupled with tools of ecological modeling, we delineated areas for fish fauna conservation and restoration in the 20 major river basins of India. To do this, we used prioritization analyses and reserve selection algorithms to derive conservation value index (CVI) and vulnerability index (VI) of the river basins. CVI was estimated using endemicity, rarity, conservation value, and taxonomic singularity, while VI was estimated using a disturbance index derived from percent geographic area of the basin under human settlements, human population density, predominant land use, and total number of exotic fish species in each basin. The two indices, CVI and VI, were converted into geo-referenced maps, and each map was super-imposed onto species richness and forest cover maps, respectively. After superimposition, areas with high CVI and low VI shade intensities were delineated for conservation, while areas with high CVI and high VI shade intensities were demarcated for restoration. In view of the importance of freshwater fish for human livelihoods and consumption, and ecosystems of India's rivers, we call for urgent attention to the conservation of their fish fauna along with restoration of their degraded habitats.

Hybrid Storage Market Assessment: A JISEA White Paper

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ericson, Sean J.; Rose, Eric; Jayaswal, Harshit

This white paper evaluates which markets are best suited for battery storage and storage hybrids and reviews regulations and incentives that support or impede the implementation of standalone storage and battery hybrids. The costs of battery storage technologies have dropped in recent years, resulting in a seven-fold increase in installed capacity over the last decade (1). These technologies offer an attractive rate of return in some locations; however, cost and regulatory barriers still limit the market for storage. Hybridizing a battery (combining the battery with a generator) can in some instances reduce total system costs and increase value compared tomore » separate installations. The fast ramping and dispatchability of a battery can complement the generator to provide services that neither battery nor generator could provide alone. Battery hybrids also benefit from some policy incentives and may be better able to meet market and regulatory requirements.« less

Cytogenetics and fluorescence in-situ hybridization in detection of hematological malignancies.

PubMed

Frenny, V J; Antonella, Z; Luisa, A; Shah, A D; Sheth, J J; Rocchi, M

2003-01-01

The technique of Fluorescence In-Situ Hybridization (FISH), a hybrid of cytogenetics and molecular biology has increased the resolution and application of cytogenetics in various neoplastic processes. In various types of leukemias, primary investigation by conventional cytogenetic [CC] technique followed by FISH has increased our understanding of the abnormal clonal formation involving different gene region. Present study is aimed to use different kinds of in-house FISH probes in various hematological malignancies and its correlation with conventional cytogenetic finding. Cytogenetic study was carried out in 360 patients either from peripheral blood or from bone marrow cells suspected for various types of leukemias. Four of 360 cases were further selected for FISH study by using different types of in-house probes, such as BAC [Bacterial Artificial Chromosome], PAC [Phague Artificial Chromosome], alphoid, PCP [Partial Chromosome Paint] and WCP [Whole Chromosome paint]. The results confirmed breakpoints of inversion 16 and del 16 in case 2 and 3 respectively. Whereas, case 1 did not confirm the cytogenetic findings of t(15;17) by PML/RARa fusion signals as multiple cell lines were involved in the patients. PCP and WCP were helpful in the identification of the marker chromosome in case 1. Telomeric and centromeric probes confirmed the cytogenetic findings of t(5;7) in case 4. We observe from this study that, in addition to the conventional cytogenetic study, FISH study provide further confirmation of chromosomal rearrangements. This facilitates our understanding of the neoplastic process more precisely for the better prognostication of the patient.

Depletion of the chloramine-T marker residue, para-toluenesulfonamide, from skin-on fillet tissue of hybrid striped bass, rainbow trout, and yellow perch

USGS Publications Warehouse

Meinertz, J.R.; Stehly, G.R.; Greseth, Shari L.; Gaikowski, M.P.; Gingerich, W.H.

2004-01-01

Waterborne exposure to n-sodium-n-chloro-p-toluenesulfonamide (chloramine-T) is an effective treatment for controlling fish mortalities caused by bacterial gill disease (BGD). Currently, data are being generated to gain United States Food and Drug Administration (FDA) approval for the use of chloramine-T in aquaculture. As part of the data required for an approval, depletion of the chloramine-T marker residue (para-toluenesulfonamide [p-TSA]) from the edible fillet tissue of exposed fish must be determined. Hybrid striped bass (Morone saxatilis??Morone chrysops; mean weight 357 g), rainbow trout (Oncorhynchus mykiss; mean weight 457 g), and yellow perch (Perca flavescens; mean weight 144 g) were exposed to 20 mg/l of chloramine-T for 60 min on 4 consecutive days (the most aggressive treatment expected for approved use in the United States). Groups of fish (n=15 or 19) were sampled immediately after the last treatment and periodically through 48 or 168 h after the treatment phase. Duplicate subsamples of skin-on fillet tissue from each fish were analyzed for p-TSA. Mean p-TSA concentrations in fillet tissue from fish sampled immediately after the last treatment were 142 ng/g (hybrid striped bass), 97 ng/g (rainbow trout), and 150 ng/g (yellow perch). Mean p-TSA concentrations at terminal sample times were 94 (168 h; hybrid striped bass), 74 (48 h; rainbow trout), and 35 ng/g (168 h; yellow perch). The half-lives of p-TSA in fillet tissue from fish near or at market size were 11.4 (hybrid striped bass), 4.3 (rainbow trout), and 3.2 days (yellow perch).

Dietary choline requirement of juvenile hybrid striped bass.

PubMed

Griffin, M E; Wilson, K A; White, M R; Brown, P B

1994-09-01

Two experiments were conducted to estimate the dietary choline requirement and to determine the effects of dietary choline on liver lipid deposition in juvenile hybrid striped bass (Monrone saxatilis x M. chrysops). Experimental diets contained 0.73 g total sulfur amino acids/100 g diet (0.47 g methionine + 0.26 g cyst(e)ine/100 g diet), thus meeting, but not exceeding, the requirement. Graded levels of choline bitartrate in Experiment 1 and choline chloride in Experiment 2 were added to the basal diet, resulting in eight dietary treatments in each experiment. Dietary treatments were 0, 250, 500, 1000, 2000, 4000, 6000 and 8000 mg choline/kg dry diet. Diets were fed for 12 and 10 wk in Experiments 1 and 2, respectively. Dietary choline concentrations significantly affected weight gain, feed efficiency, survival and total liver lipid concentrations in each experiment. Weight gain and feed efficiency were greatest in fish fed 500 mg choline/kg dry diet as choline bitartrate. Total liver lipid concentrations were variable but tended to be lowest in fish fed diets containing at least 2000 mg choline/kg diet. Survival was significantly lower in the group of fish fed 8000 mg choline/kg diet supplied by choline bitartrate. Weight gain and feed efficiency were greatest and total liver lipid concentration was lowest in groups of fish fed at least 500 mg choline/kg diet as choline chloride; survival was unaffected by dietary treatment. Therefore, choline chloride seems to be a better source of dietary choline than choline bitartrate and 500 mg choline/kg diet is adequate for maximum weight gain and prevention of increased liver lipid concentration in juvenile hybrid striped bass.

Automated processing of fluorescence in-situ hybridization slides for HER2 testing in breast and gastro-esophageal carcinomas.

PubMed

Tafe, Laura J; Allen, Samantha F; Steinmetz, Heather B; Dokus, Betty A; Cook, Leanne J; Marotti, Jonathan D; Tsongalis, Gregory J

2014-08-01

HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow. Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability. There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio >2.2) and the remaining 19 were negative (HER2:CEP17 ratio <1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5h which could then be dedicated to other lab functions. The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed

From hybrid swarms to swarms of hybrids

USGS Publications Warehouse

Stohlgren, Thomas J.; Szalanski, Allen L; Gaskin, John F.; Young, Nicholas E.; West, Amanda; Jarnevich, Catherine S.; Tripodi, Amber

2014-01-01

Science has shown that the introgression or hybridization of modern humans (Homo sapiens) with Neanderthals up to 40,000 YBP may have led to the swarm of modern humans on earth. However, there is little doubt that modern trade and transportation in support of the humans has continued to introduce additional species, genotypes, and hybrids to every country on the globe. We assessed the utility of species distributions modeling of genotypes to assess the risk of current and future invaders. We evaluated 93 locations of the genus Tamarix for which genetic data were available. Maxent models of habitat suitability showed that the hybrid, T. ramosissima x T. chinensis, was slightly greater than the parent taxa (AUCs > 0.83). General linear models of Africanized honey bees, a hybrid cross of Tanzanian Apis mellifera scutellata and a variety of European honey bee including A. m. ligustica, showed that the Africanized bees (AUC = 0.81) may be displacing European honey bees (AUC > 0.76) over large areas of the southwestern U.S. More important, Maxent modeling of sub-populations (A1 and A26 mitotypes based on mDNA) could be accurately modeled (AUC > 0.9), and they responded differently to environmental drivers. This suggests that rapid evolutionary change may be underway in the Africanized bees, allowing the bees to spread into new areas and extending their total range. Protecting native species and ecosystems may benefit from risk maps of harmful invasive species, hybrids, and genotypes.

Assessment of general health of fishes collected at selected sites in the Great Lakes Basin In 2012

USGS Publications Warehouse

Mazik, Patricia M.; Braham, Ryan P.; Hahn, Cassidy M.; Blazer, Vicki

2015-01-01

The U. S. Fish and Wildlife Service received funding through the Great Lakes Restoration Initiative (GLRI) for an Early Warning Program to detect and identify emerging contaminants and to evaluate the effects of these contaminants on fish and wildlife. The U.S. Geological Survey (WV Cooperative Fish and Wildlife Research Unit and National Fish Health Research Laboratory, Leetown Science Center) developed and implemented a biological effects monitoring protocol to assist in this program. Fish collections and measurements of biomarkers of exposure in Fall 2010 and Spring 2011 occurred at individual sites within select Areas of Concern (AOCs). They provided an assessment of the utility of the suite of biomarkers and also identified sites for more in-depth analyses. Selected areas are characterized as areas with known emerging contaminants, sensitive or listed species, areas downstream from municipal wastewater discharges or receiving waters for industrial facilities, and/or areas susceptible to agricultural or urban contamination, or harbors or ports. The results of the 2010- 2011 studies were summarized in Blazer et al. 2014 a, b, c; Braham et al. in review and Blazer et al. in review.

Seven-hour fluorescence in situ hybridization technique for enumeration of Enterobacteriaceae in food and environmental water sample.

PubMed

Ootsubo, M; Shimizu, T; Tanaka, R; Sawabe, T; Tajima, K; Ezura, Y

2003-01-01

A fluorescent in situ hybridization (FISH) technique using an Enterobacteriaceae-specific probe (probe D) to target 16S rRNA was improved in order to enumerate, within a single working day, Enterobacteriaceae present in food and environmental water samples. In order to minimize the time required for the FISH procedure, each step of FISH with probe D was re-evaluated using cultured Escherichia coli. Five minutes of ethanol treatment for cell fixation and hybridization were sufficient to visualize cultured E. coli, and FISH could be performed within 1 h. Because of the difficulties in detecting low levels of bacterial cells by FISH without cultivation, a FISH technique for detecting microcolonies on membrane filters was investigated to improve the bacterial detection limit. FISH with probe D following 6 h of cultivation to grow microcolonies on a 13 mm diameter membrane filter was performed, and whole Enterobacteriaceae microcolonies on the filter were then detected and enumerated by manual epifluorescence microscopic scanning at magnification of x100 in ca 5 min. The total time for FISH with probe D following cultivation (FISHFC) was reduced to within 7 h. FISHFC can be applied to enumerate cultivable Enterobacteriaceae in food (above 100 cells g-1) and environmental water samples (above 1 cell ml-1). Cultivable Enterobacteriaceae in food and water samples were enumerated accurately within 7 h using the FISHFC method. A FISHFC method capable of evaluating Enterobacteriaceae contamination in food and environmental water within a single working day was developed.

Genetic assessment of ornamental fish species from North East India.

PubMed

Dhar, Bishal; Ghosh, Sankar Kumar

2015-01-25

Ornamental fishes are traded with multiple names from various parts around the world, including North East India. Most are collected from the wild, due to lack of species-specific culture or breeding, and therefore, such unmanaged collection of the wild and endemic species could lead to severe threats to biodiversity. Despite many regulatory policies, trade of threatened species, including the IUCN listed species have been largely uncontrolled, due to species identification problems arising from the utilization of multiple trade names. So, the development of species-specific DNA marker is indispensable where DNA Barcoding is proved to be helpful in species identification. Here, we investigated, through DNA Barcoding and morphological assessment, the identification of 128 ornamental fish specimens exported from NE India from different exporters. The generated sequences were subjected to similarity match in BOLD-IDS as well as BLASTN, and analysed using MEGA5.2 for species identification through Neighbour-Joining (NJ) clustering, and K2P distance based approach. The analysis revealed straightforward identification of 84 specimens into 35 species, while 44 specimens were difficult to distinguish based on CO1 barcode alone. However, these cases were resolved through morphology, NJ and distanced based method and found to be belonging to 16 species. Among the 51 identified species, 14 species represented multiple trade names; 17 species belonged to threatened category. Species-level identification through DNA Barcoding along with traditional morphotaxonomy reflects its efficacy in regulating ornamental fish trade and therefore, appeals for their conservation in nature. The use of trade names rather than the zoological name created the passage for trafficking of the threatened species and demands immediate attention for sustaining wildlife conservation. Copyright © 2014 Elsevier B.V. All rights reserved.

Fluorescence in situ hybridization and qPCR to detect Merkel cell polyomavirus physical status and load in Merkel cell carcinomas.

PubMed

Haugg, Anke M; Rennspiess, Dorit; zur Hausen, Axel; Speel, Ernst-Jan M; Cathomas, Gieri; Becker, Jürgen C; Schrama, David

2014-12-15

The Merkel cell polyomavirus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). Clonal integration and tumor-specific mutations in the large T antigen are strong arguments that MCPyV is a human tumor virus. However, the relationship between viral presence and cancer induction remains discussed controversially. Since almost all studies on virus prevalence are based on PCR techniques, we performed MCPyV fluorescence in situ hybridization (FISH) on MCC to gain information about the quality of the viral presence on the single cell level. MCPyV-FISH was performed on tissue microarrays containing 62 formalin-fixed and paraffin-embedded tissue samples including all tumor grades of 42 patients. The hybridization patterns were correlated to the qPCR data determined on corresponding whole tissue sections. Indeed, MCPyV-FISH and qPCR data were highly correlated, i.e. 83% for FISH-positive and 93% for FISH-negative cores. Accordingly, the mean of the qPCR values of all MCPyV-positive cores differed significantly from the mean of the negative cores (p = 0.0076). Importantly, two hybridization patterns were definable in the MCPyV-FISH: a punctate pattern (85%) indicating viral integration, which correlated with a moderate viral abundance and a combination of the punctate with a diffuse pattern (15%), suggesting a possible coexistence of integrated and episomal virus which was associated with very high viral load and VP1 expression. Thus, MCPyV-FISH adds important information on the single cell level within the histomorphological context and could therefore be an important tool to further elucidate MCPyV related carcinogenesis. © 2014 UICC.

RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

PubMed Central

Moffitt, Jeffrey R.; Zhuang, Xiaowei

2016-01-01

Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

Identification of hemiclonal reproduction in three species of Hexagrammos marine reef fishes.

PubMed

Kimura-Kawaguchi, M R; Horita, M; Abe, S; Arai, K; Kawata, M; Munehara, H

2014-08-01

Natural hybrids between the boreal species Hexagrammos octogrammus and two temperate species Hexagrammos agrammus and Hexagrammos otakii were observed frequently in southern Hokkaido, Japan. Previous studies revealed that H. octogrammus is a maternal ancestor of both hybrids; the hybrids are all fertile females and they frequently breed with paternal species. Although such rampant hybridization occurs, species boundaries have been maintained in the hybrid zone. Possible explanations for the absence of introgressions, despite the frequent backcrossing, might include clonal reproduction: parthenogenesis, gynogenesis and hybridogenesis. The natural hybrids produced haploid eggs that contained only the H. octogrammus genome (maternal ancestor) with discarded paternal genome and generated F1 -hybrid type offspring by fertilization with the haploid sperm of H. agrammus or H. otakii (paternal ancestor). This reproductive mode was found in an artificial backcross hybrid between the natural hybrid and a male of the paternal ancestor. These findings indicate that the natural hybrids adopt hybridogenesis with high possibility and produce successive generations through hybridogenesis by backcrossing with the paternal ancestor. These hybrids of Hexagrammos represent the first hybridogenetic system found from marine fishes that widely inhabit the North Pacific Ocean. In contrast with other hybridogenetic systems, these Hexagrammos hybrids coexist with all three ancestral species in the hybrid zone. The coexistence mechanism is also discussed. © 2014 The Fisheries Society of the British Isles.

ALK status testing in non-small-cell lung carcinoma by FISH on ThinPrep slides with cytology material.

PubMed

Minca, Eugen C; Lanigan, Christopher P; Reynolds, Jordan P; Wang, Zhen; Ma, Patrick C; Cicenia, Joseph; Almeida, Francisco A; Pennell, Nathan A; Tubbs, Raymond R

2014-04-01

Oncogenic anaplastic lymphoma kinase (ALK) gene rearrangements in non-small-cell lung carcinomas (NSCLC) provide the basis for targeted therapy with crizotinib and other specific ALK inhibitors. Treatment eligibility is conventionally determined by the Food and Drug Administration-approved companion diagnostic fluorescence in situ hybridization (FISH) assay on paraffin-embedded tissue (PET). On limited samples such as fine needle aspiration-derived cytoblocks, FISH for ALK is often uninformative. FISH performed on liquid-based ThinPrep slides (ThinPrep-FISH) may represent a robust alternative. Two hundred thirty cytology samples from 217 patients with advanced NSCLC, including a consecutive series of 179 specimens, were used to generate matched ThinPrep slides and paraffin cytoblocks. The same ThinPrep slides used for cytologic diagnosis were assessed by standard ALK break-apart two-color probe FISH, after etching of tumor areas. Ultrasensitive ALK immunohistochemistry (IHC) on corresponding cytoblocks [D5F3 antibody, OptiView signal amplification] served as the reference data set. ThinPrep-FISH ALK signals were robust in 228 of 230 cases and not compromised by nuclear truncation inherent in paraffin-embedded tissue-FISH; only two samples displayed no signals. Nine of 178 informative cases (5%) in the consecutive series and 18 of 228 informative cases (7.8%) overall were ALK rearranged by ThinPrep-FISH. In 154 informative matched ThinPrep-FISH and cytoblock-IHC samples, 152 were concordant (10, 6.5% ALK status positive; 142, 92.2% ALK status negative), and two (1.3%) were ThinPrep-FISH positive but IHC negative (sensitivity 100%, specificity 98.6%, overall agreement 98.7%). Detection of ALK gene rearrangements in liquid cytology ThinPrep slides derived from patients with NSCLC can be confidently used for clinical ALK molecular testing.

Chemical and nutritional properties of channel and hybrid catfish by-products

USDA-ARS?s Scientific Manuscript database

In the past channel catfish (Ictalurus punctatus) accounted for most of the aquaculture reared fish; however, hybrid catfish (Ictalurus punctatus X Ictalurus furcatus) now account for an ever increasing percent of aquaculture reared catfish. The objective of this study was to chemically characteriz...

Assessing chronic fish health: An application to a case of an acute exposure to chemically treated crude oil.

PubMed

Mauduit, F; Domenici, P; Farrell, A P; Lacroix, C; Le Floch, S; Lemaire, P; Nicolas-Kopec, A; Whittington, M; Zambonino-Infante, J L; Claireaux, G

2016-09-01

Human alteration of marine ecosystems is substantial and growing. Yet, no adequate methodology exists that provides reliable predictions of how environmental degradation will affect these ecosystems at a relevant level of biological organization. The primary objective of this study was to develop a methodology to evaluate a fish's capacity to face a well-established environmental challenge, an exposure to chemically dispersed oil, and characterize the long-term consequences. Therefore, we applied high-throughput, non-lethal challenge tests to assess hypoxia tolerance, temperature susceptibility and maximal swimming speed as proxies for a fish's functional integrity. These whole animal challenge tests were implemented before (1 month) and after (1 month) juvenile European sea bass (Dicentrarchus labrax) had been acutely exposed (48h) to a mixture containing 0.08gL(-1) of weathered Arabian light crude oil plus 4% dispersant (Corexit© EC9500A), a realistic exposure concentration during an oil spill. In addition, experimental populations were then transferred into semi-natural tidal mesocosm ponds and correlates of Darwinian fitness (growth and survival) were monitored over a period of 4 months. Our results revealed that fish acutely exposed to chemically dispersed oil remained impaired in terms of their hypoxia tolerance and swimming performance, but not in temperature susceptibility for 1 month post-exposure. Nevertheless, these functional impairments had no subsequent ecological consequences under mildly selective environmental conditions since growth and survival were not impacted during the mesocosm pond study. Furthermore, the earlier effects on fish performance were presumably temporary because re-testing the fish 10 months post-exposure revealed no significant residual effects on hypoxia tolerance, temperature susceptibility and maximal swimming speed. We propose that the functional proxies and correlates of Darwinian fitness used here provide a useful

A comparative genomic hybridization study in a 46,XX male.

PubMed

Rigola, M Angels; Carrera, Marta; Ribas, Isabel; Egozcue, Josep; Miró, Rosa; Fuster, Carme

2002-07-01

To identify Y chromosome material in an azoospermic male with an XX karyotype. Case report. Faculty of medicine and Centro de Patologia Celular (CPC) medical center. A 33-year-old man with infertility. G-banding, fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic hybridization (CGH). FISH for X and Y chromosomes, PCR for the SRYgene and amelogenin gene in the Xp (AMGX) and (AMGY), and losses or gains with CGH. FISH analysis using X and Y chromosome-specific probes showed an X chromosome containing Y chromosome sequences on the top of the short arm; this Y chromosome region was not visible by conventional cytogenetic analysis. PCR amplification of DNA showed the presence of the sex-determining region of the Y chromosome (SRY) and the amelogenin gene in the pseudoautosomal boundary of the X chromosome (AMGX). CGH confirmed the presence of the chromosome region Yp11.2-pter and detected the presence of the two otherwise normal X chromosomes. The two Xpter (XPAR1) pseudoautosomal regions present in this XX male suggest the need to reevaluate XX males using CGH and PCR to characterize the clinical variability in XX males due to genes other than those located on the Y chromosome.

Effectiveness of FISK, an invasiveness screening tool for non-native freshwater fishes, to perform risk identification assessments in the Iberian Peninsula.

PubMed

Almeida, David; Ribeiro, Filipe; Leunda, Pedro M; Vilizzi, Lorenzo; Copp, Gordon H

2013-08-01

Risk assessments are crucial for identifying and mitigating impacts from biological invasions. The Fish Invasiveness Scoring Kit (FISK) is a risk identification (screening) tool for freshwater fishes consisting of two subject areas: biogeography/history and biology/ecology. According to the outcomes, species can be classified under particular risk categories. The aim of this study was to apply FISK to the Iberian Peninsula, a Mediterranean climate region highly important for freshwater fish conservation due to a high level of endemism. In total, 89 fish species were assessed by three independent assessors. Results from receiver operating characteristic analysis showed that FISK can discriminate reliably between noninvasive and invasive fishes for Iberia, with a threshold of 20.25, similar to those obtained in several regions around the world. Based on mean scores, no species was categorized as "low risk," 50 species as "medium risk," 17 as "moderately high risk," 11 as "high risk," and 11 as "very high risk." The highest scoring species was goldfish Carassius auratus. Mean certainty in response was above the category "mostly certain," ranging from tinfoil barb Barbonymus schwanenfeldii with the lowest certainty to eastern mosquitofish Gambusia holbrooki with the highest level. Pair-wise comparison showed significant differences between one assessor and the other two on mean certainty, with these two assessors showing a high coincidence rate for the species categorization. Overall, the results suggest that FISK is a useful and viable tool for assessing risks posed by non-native fish in the Iberian Peninsula and contributes to a "watch list" in this region. © 2013 Crown copyright This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.

Biology of extinction risk in marine fishes

PubMed Central

Reynolds, John D; Dulvy, Nicholas K; Goodwin, Nicholas B; Hutchings, Jeffrey A

2005-01-01

We review interactions between extrinsic threats to marine fishes and intrinsic aspects of their biology that determine how populations and species respond to those threats. Information is available on the status of less than 5% of the world's approximately 15 500 marine fish species, most of which are of commercial importance. By 2001, based on data from 98 North Atlantic and northeast Pacific populations, marine fishes had declined by a median 65% in breeding biomass from known historic levels; 28 populations had declined by more than 80%. Most of these declines would be sufficient to warrant a status of threatened with extinction under international threat criteria. However, this interpretation is highly controversial, in part because of a perception that marine fishes have a suite of life history characteristics, including high fecundity and large geographical ranges, which might confer greater resilience than that shown by terrestrial vertebrates. We review 15 comparative analyses that have tested for these and other life history correlates of vulnerability in marine fishes. The empirical evidence suggests that large body size and late maturity are the best predictors of vulnerability to fishing, regardless of whether differences among taxa in fishing mortality are controlled; there is no evidence that high fecundity confers increased resilience. The evidence reviewed here is of direct relevance to the diverse criteria used at global and national levels by various bodies to assess threat status of fishes. Simple life history traits can be incorporated directly into quantitative assessment criteria, or used to modify the conclusions of quantitative assessments, or used as preliminary screening criteria for assessment of the ∼95% of marine fish species whose status has yet to be evaluated either by conservationists or fisheries scientists. PMID:16243696

Biology of extinction risk in marine fishes.

PubMed

Reynolds, John D; Dulvy, Nicholas K; Goodwin, Nicholas B; Hutchings, Jeffrey A

2005-11-22

We review interactions between extrinsic threats to marine fishes and intrinsic aspects of their biology that determine how populations and species respond to those threats. Information is available on the status of less than 5% of the world's approximately 15500 marine fish species, most of which are of commercial importance. By 2001, based on data from 98 North Atlantic and northeast Pacific populations, marine fishes had declined by a median 65% in breeding biomass from known historic levels; 28 populations had declined by more than 80%. Most of these declines would be sufficient to warrant a status of threatened with extinction under international threat criteria. However, this interpretation is highly controversial, in part because of a perception that marine fishes have a suite of life history characteristics, including high fecundity and large geographical ranges, which might confer greater resilience than that shown by terrestrial vertebrates. We review 15 comparative analyses that have tested for these and other life history correlates of vulnerability in marine fishes. The empirical evidence suggests that large body size and late maturity are the best predictors of vulnerability to fishing, regardless of whether differences among taxa in fishing mortality are controlled; there is no evidence that high fecundity confers increased resilience. The evidence reviewed here is of direct relevance to the diverse criteria used at global and national levels by various bodies to assess threat status of fishes. Simple life history traits can be incorporated directly into quantitative assessment criteria, or used to modify the conclusions of quantitative assessments, or used as preliminary screening criteria for assessment of the approximately 95% of marine fish species whose status has yet to be evaluated either by conservationists or fisheries scientists.

Assessment of Habitat, Fish Communities, and Streamflow Requirements for Habitat Protection, Ipswich River, Massachusetts, 1998-99

USGS Publications Warehouse

Armstrong, David S.; Richards, Todd A.; Parker, Gene W.

2001-01-01

The relations among stream habitat, fish communities, and hydrologic conditions were investigated in the Ipswich River Basin in northeastern Massachusetts. Data were assessed from 27 sites on the mainstem of the Ipswich River from July to September 1998 and from 10 sites on 5 major tributaries in July and August 1999. Habitat assessments made in 1998 determined that in a year with sustained streamflow for most of the summer, the Ipswich River contains diverse, high-quality aquatic habitat. Channel types are predominantly low gradient glides, pools, and impoundments, with a sandy streambed and a forest or shrub riparian zone. Features that provide fish habitat are located mostly along stream margins; these features include overhanging brush, undercut banks, exposed roots, and woody debris. These habitat features decrease in availability to aquatic communities with declining streamflows and generally become unavailable after streamflows drop to the point where the edge of water recedes from the stream banks.The mainstem and tributaries were sampled to determine fish species composition, relative abundance, and length frequency. Fish sampling indicates that the fish community in the Ipswich River is currently a warm-water fish community dominated by pond-type fish. However, historical temperature data, and survival of stocked trout in the mainstem Ipswich into late summer of 1998, indicate that the Ipswich River potentially could support cold-water fish species if adequate flows are maintained. Dominant fish species sampled in the mainstem Ipswich River were redfin pickerel (Esox americanus), American eel (Anguilla rostrata), and pumpkinseed (Lepomis gibbosus), which together represented 41, 22, and 10 percent, respectively, of 4,745 fish sampled. The fish communities of the mainstem and tributaries contained few fluvial-dependent or fluvial-specialist species (requiring flow), and were dominated by macrohabitat generalists (tolerant of low-flow, warm-water, and

Impact of fishing and stocking practices on Coregonid diversity

USGS Publications Warehouse

Anneville, Orlane; Lasne, Emilien; Guillard, Jean; Eckmann, Reiner; Stockwell, Jason D.; Gillet, Christian; Yule, Daniel

2015-01-01

Fish species diversity can be lost through interacting stressors including habitat loss, stocking and overfishing. Although a multitude of stressors have played a role in the global decline of coregonid (Coregonus spp.) diversity, a number of contemporary studies have identified habitat loss stemming from eutrophication as the primary cause. Unfortunately, reconstructing the role of fishing and stocking practices can be difficult, because these records are incomplete or appear only in hard-to-access historic grey literature. Based on an illustrative set of historic and contemporary studies, we describe how fisheries management practices may have contributed to coregonid diversity loss in European and North American lakes. We provide case studies examining how fishing and stocking may reduce coregonid diversity through demographic decline and introgressive hybridization. In some lakes, fisheries management practices may have led to a loss of coregonid diversity well before issues with habitat degradation manifested. Our review suggests that fish conservation policies could beneficially consider the relative importance of all stressors, including management practices, as potential drivers of diversity loss.

Assessment of the effects rejections of feed fish on water resources.: (Ouedoumerrbia, Morocco)

NASA Astrophysics Data System (ADS)