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Sample records for flounder pseudopleuronectes americanus

  1. Assessment of Dredge-induced Sedimentation Effects on Winter Flounder (pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  2. Assessment of Dredging-Induced Sedimentation Effects on Winter Flounder (Pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  3. Liver lesions in winter flounder (Pseudopleuronectes americanus) from Jamaica Bay, New York: Indications of environmental degradation

    USGS Publications Warehouse

    Augspurger, T.P.; Herman, R.L.; Tanacredi, J.T.; Hatfield, J.S.

    1994-01-01

    Liver sections of winter flounder (Pseudopleuronectes americanus) collected from Jamaica Bay and Shinnecock Bay, New York, in 1989, were examined microscopically to determine the pervasiveness of liver lesions observed previously in Jamaica Bay winter flounder. Neoplastic lesions were not detected in fish from Jamaica Bay or the Shinnecock Bay reference site. Twenty-two percent of Jamaica Bay winter flounder examined (n = 103) had unusual vacuolization of hepatocytes and biliary pre-ductal and ductal cells (referred to hereafter as the vacuolated cell lesion). The lesion, identical to that found in 25% of Jamaica Bay winter flounder examined in 1988, has previously been identified in fishes taken from highly polluted regions of the Atlantic coast (e.g., Boston Harbor, Massachusetts, and Black Rock Harbor, Connecticut). Prevalence of the vacuolated cell lesion in winter flounder from Jamaica Bay was significantly greater (p < 0.0001) than in 102 specimens collected from Shinnecock Bay. Current scientific literature indicates vacuolated hepatocytes and cholangiocytes are chronically injured and that the extent of their deformity is consistent with the action of a hepatotoxicant. The high prevalence of vacuolated hepatocytes in Jamaica Bay winter flounder and absence of the lesion in flounder from reference sites strongly supports the hypothesis that this impairment is a manifestation of a toxic condition in at least some portions of Jamaica Bay.

  4. Windowpane flounder (Scophthalmus aquosus) and winter flounder (Pseudopleuronectes americanus) responses to cold temperature extremes in a Northwest Atlantic estuary

    NASA Astrophysics Data System (ADS)

    Wilber, Dara H.; Clarke, Douglas G.; Alcoba, Catherine M.; Gallo, Jenine

    2016-01-01

    The effect of climate variability on flatfish includes not only the effects of warming on sensitive life history stages, but also impacts from more frequent or unseasonal extreme cold temperatures. Cold weather events can affect the overwintering capabilities of flatfish near their low temperature range limits. We examined the responses of two flatfish species, the thin-bodied windowpane (Scophthalmus aquosus) and cold-tolerant winter flounder (Pseudopleuronectes americanus), to variable winter temperatures in a Northwest Atlantic estuary using abundance and size data collected during a monitoring study, the Aquatic Biological Survey, conducted from 2002 to 2010. Winter and spring abundances of small (50 to 120 mm total length) juvenile windowpane were positively correlated with adult densities (spawning stock) and fall temperatures (thermal conditions experienced during post-settlement development for the fall-spawned cohort) of the previous year. Windowpane abundances in the estuary were significantly reduced and the smallest size class was nearly absent after several consecutive years with cold (minimum temperatures < 1 °C) winters. Interannual variation in winter flounder abundances was unrelated to the severity of winter temperatures. A Paulik diagram illustrates strong positive correlations between annual abundances of sequential winter flounder life history stages (egg, larval, Age-1 juvenile, and adult male) within the estuary, reflecting residency within the estuary through their first year of life. Temperature variables representing conditions during winter flounder larval and post-settlement development were not significant factors in multiple regression models exploring factors that affect juvenile abundances. Likewise, densities of predators known to consume winter flounder eggs and/or post-settlement juveniles were not significantly related to interannual variation in winter flounder juvenile abundances. Colder estuarine temperatures through the

  5. Effects of cooking on levels of PCBs in the fillets of winter flounder (Pseudopleuronectes americanus)

    SciTech Connect

    Poston, T.M.; Durell, G.S.; Koczwara, G.; Spellacy, A.M.

    1995-08-01

    The Pacific Northwest Laboratory and Battelle Ocean Sciences performed a study to determine the effect of cooking on polychlorinated biphenyl (PCB) levels in the fillets of winter flounder (Pseudopleuronectes americanus). Broiling, pan frying, and deep frying in oil were tested on fillets from 21 fish collected from New Bedford Harbor, Massachusetts, on February 21, 1991. The evaluation involved estimating the change in PCB concentrations using a mass-balance approach that factored the change in fillet weight resulting from cooking with the changes in PCB concentration expressed on a precooked wet-weight basis. Deep frying in oil resulted in a 47% reduction in total PCB levels in fillet tissue. Additionally, deep frying caused a 40% reduction in fillet mass. Pan frying and broiling resulted in statistically in insignificant increases in total PCB levels of 15% and 17%, respectively. Fillet mass reductions resulting from pan frying and broiling were 7% and 15%, respectively. The effects of cooking on 18 individual congeners generally paralleled the results observed for total PCB. All 18 congeners were significantly reduced by deep frying. Congener Cl{sub 2}(08) also was significantly reduced by either pan frying. Congeners Cl{sub 5}(105) and Cl{sub 5}(118) showed apparent significant increases in concentrations following pan frying. Congeners Cl{sub 5}(105), Cl{sub 5}(118), and C1{sub 6}(138) showed significant increases in concentration following broiling.

  6. A New PCR-Based Method Shows That Blue Crabs (Callinectes sapidus (Rathbun)) Consume Winter Flounder (Pseudopleuronectes americanus (Walbaum))

    PubMed Central

    Collier, Jackie L.; Fitzgerald, Sean P.; Hice, Lyndie A.; Frisk, Michael G.; McElroy, Anne E.

    2014-01-01

    Winter flounder (Pseudopleuronectes americanus) once supported robust commercial and recreational fisheries in the New York (USA) region, but since the 1990s populations have been in decline. Available data show that settlement of young-of-the-year winter flounder has not declined as sharply as adult abundance, suggesting that juveniles are experiencing higher mortality following settlement. The recent increase of blue crab (Callinectes sapidus) abundance in the New York region raises the possibility that new sources of predation may be contributing to juvenile winter flounder mortality. To investigate this possibility we developed and validated a method to specifically detect winter flounder mitochondrial control region DNA sequences in the gut contents of blue crabs. A survey of 55 crabs collected from Shinnecock Bay (along the south shore of Long Island, New York) in July, August, and September of 2011 showed that 12 of 42 blue crabs (28.6%) from which PCR-amplifiable DNA was recovered had consumed winter flounder in the wild, empirically supporting the trophic link between these species that has been widely speculated to exist. This technique overcomes difficulties with visual identification of the often unrecognizable gut contents of decapod crustaceans, and modifications of this approach offer valuable tools to more broadly address their feeding habits on a wide variety of species. PMID:24454797

  7. Cadmium inhibition of L-alanine transport into renal brush border membrane vesicles isolated from the winter flounder (Pseudopleuronectes americanus)

    SciTech Connect

    Bevan, C.; Kinne-Saffran, E.; Foulkes, E.C.; Kinne, R.K. )

    1989-12-01

    Using isolated brush border membrane vesicles from the kidney of the winter flounder (Pseudopleuronectes americanus), we have studied the effect of cadmium on L-alanine transport. Pretreatment of vesicles with 0.1 mM Cd{sup 2+} resulted in inhibition of L-alanine uptake in the presence of a NaCl (but not KCl) gradient. Inhibition was due to a specific interaction with the sodium-alanine cotransport system and not a change in the driving forces for alanine transport, since Cd{sup 2+} did not affect sodium-dependent D-glucose uptake. The effect of Cd{sup 2+} on Na{sup +}-alanine cotransport showed mixed-type inhibition which is only partially reversible by EDTA. Cd{sup 2+} uptake itself was shown to be time and temperature dependent, resulting in binding to both sides of the membrane. No direct correlation was possible between inhibition of L-alanine transport and the amount of Cd{sup 2+} taken up by the membranes. Nevertheless, the striking time dependence of the effect of Cd{sup 2+} on sodium-dependent L-alanine uptake and the inability of EDTA to reverse the inhibitory action of Cd{sup 2+} suggest that Cd{sup 2+} inhibits Na+-alanine cotransport at the cytoplasmic side of the membrane.

  8. Comparative reproductive success of winter flounder in Long Island Sound: A three-year study (biology, biochemistry, and chemistry). [Pseudopleuronectes americanus

    SciTech Connect

    Nelson, D.A.; Miller, J.E.; Rusanowsky, D.; Greig, R.A.; Sennefelder, G.R.; Mercaldo-Allen, R.; Kuropat, C.; Gould, E.; Thurberg, F.P.; Calabrese, A. )

    1991-09-01

    In a 3-yr study, late prespawning winter flounder (Pseudopleuronectes americanus) were collected from various stations in Long Island Sound (three of them heavily urbanized) and spawned in the laboratory. For comparative purposes, flounder from two sites in the Boston Harbor area were similarly treated in 1987 and 1988. Of the stations in Long Island Sound, New Haven Harbor alone consistently produced low percent viable hatch and small larvae. Boston Harbor produced the smallest larvae of all sites. There were no significant station-to-station differences in lipid utilization during larval development; yolk reserves at stations showing a low percent viable hatch, small larvae, and low embryonic development rate were probably used in part for stress metabolism. No significant differences in concentrations of polychlorinated biphenyls for collections from Long Island Sound were found either in livers of spawned fish, in sediments, or in eggs of winter flounder. The very low metal concentrations in winter flounder eggs showed no relation to the degree of metal contamination found at stations in Long Island Sound and Boston Harbor.

  9. Predator-prey relations between age-1+ summer flounder (Paralichthys dentatus, Linnaeus) and age-0 winter flounder (Pseudopleuronectes americanus, Walbaum): predator diets, prey selection, and effects of sediments and macrophytes.

    PubMed

    Manderson; Phelan; Stoner; Hilbert

    2000-08-23

    Laboratory experiments and weekly trammel net surveys in the Navesink River, New Jersey (USA) were used to examine the predator-prey interaction between age-1+ summer flounder (Paralichthys dentatus) and age-0 winter flounder (Pseudopleuronectes americanus). Winter flounder (24-67 mm TL) were the dominant piscine prey of summer flounder (n=95, 252-648 mm TL) collected in trammel nets. We observed a temporal shift in summer flounder diets from sand shrimp (Crangon septemspinosa) and winter flounder, dominant during June and early July, to blue crabs (Callinectes sapidus) and other fishes (primarily Atlantic silversides, Menidia menidia and Atlantic menhaden, Brevortia tyrannus) later in the summer. Variations in prey selection appeared to be related to changes in the spatial distribution of predators and spatio-temporal variation in prey availability. In laboratory experiments, summer flounder (271-345 mm total length, TL) preferred demersal winter flounder to a pelagic fish (Atlantic silversides) and a benthic invertebrate (sand shrimp) prey, and the vulnerability of winter flounder increased with increasing prey body size from 20 to 90 mm TL. Experiments testing habitat effects showed that mortality of winter flounder in three different size classes (20-29, 40-49, 60-69 mm TL) was not influenced by sediment grain sizes permitting differential burial of the prey. However, vegetation enhanced survival, with fish suffering lower mortality in eelgrass (Zostera marina, 15+/-0.04%) than in sea lettuce (Ulva lactuca, 38+/-0.04%) or bare sand (70+/-0.07%) when the macrophytes were planted to produce similar leaf surface areas (5000 cm(2) m(-2)). Prey vulnerability appeared to be related to the role of vision in the predator's attack strategy and prey activity levels. PMID:10958899

  10. Construction of a subtractive library from hexavalent chromium treated winter flounder (Pseudopleuronectes americanus) reveals alterations in non-selenium glutathione peroxidases.

    PubMed

    Chapman, Laura M; Roling, Jonathan A; Bingham, Lacey K; Herald, Matt R; Baldwin, William S

    2004-04-14

    Chromium is released during several industrial processes and has accumulated in some estuarine areas. Its effects on mammals have been widely studied, but relatively little information is available on its effects on fish. Gene expression changes are useful biomarkers that can provide information about toxicant exposure and effects, as well as the health of an organism and its ability to adapt to its surroundings. Therefore, we investigated the effects of Cr(VI) on gene expression in the sediment dwelling fish, winter flounder (Pseudopleuronectes americanus). Winter flounder ranging from 300 to 360 g were injected i.p. with Cr(VI) as chromium oxide at 25 microg/kg chromium in 0.15N KCl. Twenty-four hours following injections, winter flounder were euthanized with MS-222 and the livers were excised. Half of the livers were used to make cytosol and the other half were used to isolate mRNA for subtractive hybridization. Subtractive clones obtained were spotted onto nylon filters, which revealed several genes with potentially altered expression due to Cr(VI), including an alpha class GST, 1-Cys peroxiredoxin (a non-selenium glutathione peroxidase), a P-450 2X subfamily member, two elongation factors (EF-1 gamma and EF-2), and complement component C3. Semi-quantitative RT-PCR was performed and confirmed that Cr(VI) down-regulated complement component C3, an EST, and two potential glutathione peroxidases, GSTA3 and 1-Cys peroxiredoxin. In addition, cytosolic GSH peroxidase activity was reduced, and silver stained SDS-PAGE gels from glutathione-affinity purified cytosol demonstrated that a 27.1 kDa GSH-binding protein was down-regulated greater than 50%. Taken together, Cr(VI) significantly altered the expression of several genes including two potential glutathione peroxidases in winter flounder. PMID:15003702

  11. Time course of oocyte development in winter flounder Pseudopleuronectes americanus and spawning seasonality for the Gulf of Maine, Georges Bank and southern New England stocks.

    PubMed

    Press, Y K; McBride, R S; Wuenschel, M J

    2014-08-01

    Winter flounder Pseudopleuronectes americanus were collected at monthly intervals from December 2009 to May 2011, to describe the pattern and seasonality of oocyte development, including: (1) the group-synchronous transition from primary to secondary oocytes that initiates immediately after spawning, (2) the slow (months) development of vitellogenic oocytes followed by the rapid (weeks) maturation of oocytes, (3) the synchronous nature of mature oocytes ovulating, but the discrete releases of benthic eggs in batches, (4) the protracted (months) degradation of postovulatory follicles and (5) the occurrence of follicular atresia. Although fish were collected across only c. 2° latitudinal range, the spawning season was c. 1 month later in the Gulf of Maine (GOM) than on Georges Bank and in southern New England. This is probably due to lower temperatures in the GOM. These stock-specific data regarding the time course of oogenesis are of practical value. This information is discussed in relation to measuring and interpreting elements of reproductive potential such as maturation, skipped spawning and fecundity, the response of reproductive traits by this widely distributed species to changing climate and the response by this common, marine-estuarine species to urbanization, particularly environmental pollutants and dredging.

  12. Predator prey size relationship between Pseudopleuronectes americanus and Carcinus maenas

    NASA Astrophysics Data System (ADS)

    Fairchild, E. A.; Howell, W. H.

    2000-10-01

    Young-of-year flatfish grow through a series of critical periods in which they are vulnerable to different predators, including decapod crustaceans. The purpose of this study was to determine if winter flounder, Pseudopleuronectes americanus, were vulnerable to one such decapod, the green crab, Carcinus maenas, and to determine if vulnerability differed between wild and cultured fish. To examine the predator-prey size relationship, an experiment was conducted in which six cultured and three wild winter flounder size class treatments were tested against six crab size class treatments. Flounder of all size classes were preyed on by all size classes of green crabs; however, mortality was highest when the largest crabs were matched with the smallest flounder. The number of flounder killed per day was significantly higher (31%) in winter flounder <20 mm compared to all other larger fish size classes (4-8%). Additionally, these fish were attacked at a faster rate than any other fish size class. For the 31-60 mm fish size classes tested, more wild fish (11%) were killed per day by crabs than cultured fish (6.3%). These results suggest that in a winter flounder stock enhancement program, only fish >20 mm should be released to promote post-release survival.

  13. Pharmacokinetics of morphine in fish: winter flounder (Pseudopleuronectes americanus) and seawater-acclimated rainbow trout (Oncorhynchus mykiss).

    PubMed

    Newby, Nathalie C; Mendonça, Paula C; Gamperl, Kurt; Stevens, E Don

    2006-07-01

    We made a single intraperitoneal (IP) injection of morphine sulfate (40 mg/kg) into winter flounder and seawater acclimated rainbow trout at 10 degrees C and then followed its disposition by measuring the change in plasma morphine concentration for 100 h using a morphine specific ELISA. Disposition also was followed for 6h after a single IV injection of 7.5mg morphine sulfate in winter flounder. Plasma morphine reached a maximum within an hour post-injection IP and then decreased in a bi-exponential fashion with a rapid distribution phase followed by a slower elimination phase. The disposition was slower in flounder than in trout even though the fish were held at the same temperature. For example, plasma clearance was 76 mL h(-)(1) kg(-)(1) in the flounder but was almost twice as much in the trout (153 mL h(-)(1) kg(-)(1)) and mean residence time was 27.9h in the flounder but was 7.0 h in the trout. The present study is the first comprehensive pharmacokinetic analysis for any analgesic in an ectotherm, and our results show that: 1) significant intra-specific variation exists between fishes: and 2) the disposition of morphine in fish is approximately one order of magnitude slower than it is in mammals. These differences may be due in part to mass specific differences in cardiac output.

  14. Winter flounder (Pseudopleuronectes americanus) estuarine habitat use and the association between spring temperature and subsequent year class strength

    NASA Astrophysics Data System (ADS)

    Wilber, Dara H.; Davis, David; Clarke, Douglas G.; Alcoba, Catherine J.; Gallo, Jenine

    2013-11-01

    Winter flounder habitat use in New York/New Jersey Harbor was examined through a long-term (2002-2010) bottom trawl sampling program (Aquatic Biological Survey) in which 5234 winter flounder were captured in 2069 samples collected at approximately 26 stations throughout the harbor. Interannual variability in catch-per-unit-effort (CPUE) primarily was attributable to fluctuations in Year-1 juvenile CPUE, which was positively correlated with total annual egg abundances from the previous year. Adult male CPUE during the spawning season was positively correlated with total egg abundances of the same year, whereas adult female CPUE was unrelated to annual egg abundances in the harbor. Annual variation in adult male densities in the harbor during the spawning season reflects the intensity of estuarine spawning activity, whereas adult female densities may include non-reproductive, foraging individuals. Seasonal fluctuations in condition indices reflected energy use during the spawning season, with relatively high condition in January, reduced levels in March and April, and elevated condition again in May. Adult CPUE peaked in April, coincident with the critical feeding period that follows spawning. Mean April water temperatures were positively correlated with egg abundances the following year and Year-1 juvenile CPUE two years later. A similar correlation between April temperatures and Year-1 juvenile abundances two years later was demonstrated using published data for winter flounder collected in Niantic Bay, CT. Higher April water temperatures may enhance benthic secondary production during the critical feeding period, and thus increase prey availability for foraging adults that need to restore energy reserves in order to reproduce the following year. A direct examination of benthic secondary production and variation in winter flounder estuarine foraging and subsequent spawning activity is needed to more fully understand this relationship.

  15. Long-term assessment of settlement and growth of juvenile winter flounder ( Pseudopleuronectes americanus) in New Jersey estuaries

    NASA Astrophysics Data System (ADS)

    Sogard, Susan M.; Able, Kenneth W.; Hagan, Stacy M.

    2001-06-01

    We examined patterns of metamorphosis, settlement, and growth for young-of-the-year winter flounder over a ten year period (1990-1999) in four estuaries spanning the coast of New Jersey, USA, with the objective to determine the spatial and temporal coherence in these processes. We hypothesised that these processes could be influenced by large-scale climatic factors such as temperature and small-scale, local factors such as predator and prey abundances and differences associated with the localised population structure of winter flounder. Newly settled juveniles were collected in late May/early June each year. Increment counts on sagittal otoliths were used to determine the date of metamorphosis, and increment widths were used to estimate growth rates at two times, just after settlement and just prior to capture. A high level of consistency in spatial patterns among four individual estuaries was evident, and was largely driven by the northernmost site, Sandy Hook, which had higher fish densities and later metamorphosis than the three other sites, and faster growth rates than the two mid-coast sites (Barnegat Bay and Great Bay). In contrast to spatial patterns, consistent temporal patterns were evident only in the timing of metamorphosis, which was delayed in colder years at all sites. Weekly plankton sampling over the same ten year period at one site (Great Bay) yielded winter flounder larvae during a relatively narrow period in spring, with a later peak of abundance in colder years, in agreement with back-calculated dates of metamorphosis for settled juveniles. These patterns suggest that the timing of settlement is driven by large-scale climatic factors that influence the timing of spawning and/or larval development. Interannual variability in densities and growth rates, however, appeared to be independent at each site, indicating control by local environmental factors or genetic differences among local populations. The spatial consistency of growth rates suggests

  16. Stable Carbon and Oxygen Isotope Ratios of Otoliths Differentiate Winter Flounder (Pseudopleuonectes americanus) Habitats

    EPA Science Inventory

    Stable carbon (13C) and oxygen (18O) isotope ratios were measured in otoliths of juvenile winter flounder (Pseudopleuronectes americanus) collected from 18 nursery areas along the coast of Rhode Island, USA. Samples were obtained during June and July of 2002 from locations tha...

  17. Latitudinal and stock-specific variation in size- and age-at-maturity of female winter flounder, Pseudopleuronectes americanus, as determined with gonad histology

    NASA Astrophysics Data System (ADS)

    McBride, Richard S.; Wuenschel, Mark J.; Nitschke, Paul; Thornton, Grace; King, Jeremy R.

    2013-01-01

    Female winter flounder were examined using gonad histology to determine the adequacy of routine macroscopic maturity classification methods and to determine the spatial variation in size and age of maturity in U.S. waters. Sampling occurred in spring and autumn, which was adequate to collect immature, mature, spawning-active, and non-active females. Females were collected in coastal waters from Delaware Bay, USA, to the Scotian Shelf, Canada, including in Long Island Sound and on Georges Bank, which covered all U.S. stock areas. Mature fish spawned in spring, when gonads comprised up to 30% of the total body weight. Direct comparisons of maturity assignment by macroscopic versus microscopic methods demonstrated that both schemes are compatible, but the more cost-effective macroscopic method had trouble distinguishing larger immature from smaller resting females. Spatial comparisons, using gonad histology only, supported the existence of three stocks in U.S. waters, but also revealed significant variation in age at maturity within the two coastal stocks. Age-at-maturity was more variable than size-at-maturity, which is consistent with known stock-specific patterns of growth rates and a postulated life history tradeoff to delay maturity until a size threshold is reached. The within-stock variation in median age at maturity, about one year for coastal stocks, recommends further investigation of using static, stock-specific maturity ogives to calculate reference points for management.

  18. Multidisciplinary assessment of pollution at three sites in Long Island Sound. [Pseudopleuronectes americanus

    SciTech Connect

    Gronlund, W.D.; Sinlam Chan; McCain, B.B.; Clark, R.C. Jr.; Myers, M.S.; Stein, J.E.; Brown, D.W.; Landahl, J.T.; Krahn, M.M.; Varanasi, U. )

    1991-09-01

    Winter flounder (Pseudopleuronectes americanus) were sampled from three sites located near Norwalk, New Haven, and Niantic, Connecticut, in Long Island Sound during February 1987, to evaluate the degree of chemical contamination and to determine possible effects of contaminant exposure. At each site, sediment ad infaunal invertebrates were also collected and analyzed for trace metals and organic chemicals. Specimens of liver and kidney from winter flounder were examined for histopathological conditions, including the presence of macrophage aggregates in liver tissue. Liver samples were also analyzed for DNA damage (i.e., the formation of adducts between DNA and chemical contaminants). Blood samples were collected and analyzed for erythrocyte micronuclei. The sampling site near New Haven was determined to be the most affected site, from the standpoints of greater chemical contamination and possible effects on winter flounder. Concentrations of aromatic hydrocarbons (AHs) and polychlorinated biphenyls (PCBs) were highest in sediment from this site, and the highest prevalences of the histopathological changes and DNA alterations were also found in the livers of winter flounder from this site. No differences in the concentrations of contaminants in fish or in frequencies of erythrocyte micronuclei in fish blood were found between sites. None of the sites samples had contaminant levels or prevalences of lesions as high as previously found at other East Coast locations (e.g., Boston Harbor, Massachusetts, Raritan Bay, New York). Overall, the results indicate moderate levels of pollution at two of the urban sites in Long Island Sound and provide a framework for expanded studies to better define the extent and impact of chemical pollution in Long Island Sound.

  19. Temporal trends in nitrogen isotope ratios of winter flounder collected from Rhode Island coastal systems

    EPA Science Inventory

    Nitrogen isotope ratios (15N) were measured in muscle tissue of juvenile winter flounder, Pseudopleuronectes americanus, collected from several estuarine systems along the coast of Rhode Island, USA, including Narragansett Bay, Narrow River and three coastal lagoons. Fish collect...

  20. Saptial and Temporal in Stable Carbon and Oxygen Isotope Ratios of Juvenile Winter Flounder Otoliths From Selected Nursery Areas

    EPA Science Inventory

    Winter flounder (Pseudopleuronectes americanus) populations have supported large commercial and recreational fisheries along the coast of New England. In recent years, however, the population of this important species has declined precipitously in some areas, especially Narragan...

  1. Novel microsatellite marker development from the unassembled genome sequence data of the marbled flounder Pseudopleuronectes yokohamae.

    PubMed

    Minegishi, Yuki; Ikeda, Minoru; Kijima, Akihiro

    2015-12-01

    Various genome-scale data have been increasingly published in diverged species, but they can be reused for other purposes by re-analyzing in other ways. As a case study to utilize the published genome data, we developed microsatellite markers from the genome sequence data (assembled contigs and unassembled reads) of the marbled flounder Pseudopleuronectes yokohamae. No microsatellites were identified in the contig sequences, whereas the computer software found 781,773 sequences containing microsatellites with di- to hexa-nucleotide motif in the unassembled reads. For 86,732 unique sequences among them, a total of 331,368 primer pairs were designed. Screening based on PCR amplification, polymorphisms and accurate genotyping resulted in sixteen primer sets, which were later characterized using 45 samples collected in Onagawa Bay, Miyagi, Japan. The presence of null alleles was suggested at four loci in the studied population but no evidence of allelic dropout was found. The observed number of alleles and heterozygosity was 2-20 and 0-0.88889, respectively, indicating polymorphisms and usefulness for population genetic analyses of this species. In addition, a large number of the microsatellite primers developed in this study are potentially applicable also for kinship estimation, individual fingerprint and linkage map construction.

  2. RELATIONSHIPS BETWEEN HABITAT QUALITY AND DENSITY OF JUVENILE WINTER FLOUNDER

    EPA Science Inventory

    We used a digital video camera mounted to a 1-m beam trawl together with an attached continuous recording YSI sonde and GPS unit to quantify juvenile winter flounder (Pseudopleuronectes americanus) densities and fish habitat. The YSI sonde measured temperature, salinity, dissolve...

  3. Carbon and nitrogen isotope ratios of juvenile winter flounder as indicators of inputs to estuarine systems

    EPA Science Inventory

    Stable carbon and nitrogen isotope ratios were measured in the muscle tissues of young-of-the-year (YOY) winter flounder, Pseudopleuronectes americanus, collected from several estuarine systems along the coast of Rhode Island, USA. These systems included three coastal lagoons (Ni...

  4. USE OF VIDEO TO ACCESS JUVENILE WINTER FLOUNDER DENSITIES AND HABITATS

    EPA Science Inventory

    We used a digital video camera mounted to a 1-m beam trawl together with an attached continuous recording YSI sonde and a GPS unit to quantify juvenile winter flounder (Pseudopleuronectes americanus) densities and fish habitat in Narragansett Bay, RI. The YSI sonde measured te...

  5. Nitrogen Isotope Ratios of Juvenile Winter Flounder as an Indicator of Anthropogenic Nitrogen Inputs to Estuarine Systems

    EPA Science Inventory

    Nitrogen isotope ratios (15N) were measured in muscle tissue of juvenile winter flounder, Pseudopleuronectes americanus, collected from several estuarine systems (lagoons, river, bay) along the coast of Rhode Island, USA over a three-year period. Significant differences i...

  6. IMPACT OF ELEVATED TEMPERATURE ON THE GROWTH, SURVIVAL, AND TROPHIC DYNAMICS OF WINTER FLOUNDER LARVAE: A MESOCOSM STUDY

    EPA Science Inventory

    Winter flounder (Pseudopleuronectes americanus) is a dominant commercial fish in Narragansett Bay, Rhode Island, and yet factors controlling its recruitment remain unclear. An experiment was conducted with six 13-m3 land-based mesocosms (5 m deep) from February to April 1997 to a...

  7. A Case Study of the Effects of Dredging in Narrgansett Bay (RI, USA) on Winter Flounder Eggs

    EPA Science Inventory

    Seasonal dredging constraints have been established in several northeastern U.S. estuaries with the intent to protect winter flounder (Pseudopleuronectes americanus). A major source of concern is impacts on demersal eggs due to elevated sedimentation rates during the winter-earl...

  8. USE OF A RHODE ISLAND SALT POND BY JUVENILE WINTER FLOUNDER, PSEUDOPLEURONECTES AMERICANUS

    EPA Science Inventory

    We used a 1.75 m2 drop ring sampler in June and July of 2000 to quantify populations of juvenile flatfishes and other small nekton in Ninigret Pond, Rhode Island. The drop sampler was deployed in approximately 1 m of water from a boom mounted on the bow of a small boat. Abundance...

  9. ASSESSING HABITAT QUALITY OF MOUNT HOPE BAY AND NARRAGANSETT BAY USING GROWTH, RNA:DNA, AND FEDDING HABITS OF CAGED JUVENILE WINTER FLOUNDER

    EPA Science Inventory

    Somatic growth rates, RNA:DNA, and feeding habits of juvenile Pseudopleuronectes americanus (Winter Flounder) were used to asses small-scale spatio-temporal variations in the habitat quality of Mount Hope Bay and Narragan-sett Bay, RI. Three successive caging experiments (14–16 d...

  10. Two species of commercial flatfish, winter flounder, Pleuronectes americanus, and American plaice, Hippoglossoides platessoides, as sentinels of environmental pollution.

    PubMed

    Khan, R A

    2010-08-01

    Two species of marketable-size flatfish, winter flounder (Pleuronectes americanus) and American plaice (Hippoglossoides platessoides) from a fjord, were examined as sentinels for the effects of effluent discharged by a pulp and paper mill in Newfoundland, Canada. Reference samples for comparison were obtained from a pristine fjord. Condition factor, lymphocyte levels and the abundance of two intestinal parasites were significantly greater in flounder and plaice sampled at the reference site while macroscopic and microscopic lesions, and hepatic somatic indices were greater in flatfish taken from the contaminated site. These results suggest that both species of commercial flatfish are useful as sentinels for studying environmental pollution.

  11. The Effect of Elevated CO2 on the Growth and Food Consumption of Juvenile Winter Flounder Pseudopleuronectes Americanus

    EPA Science Inventory

    Increasing levels of atmospheric carbon dioxide are causing changes in seawater chemistry in the world’s oceans. In estuarine waters, atmospheric CO2 exacerbates already declining pH due to high productivity and respiration caused by cultural eutrophication. These two sources o...

  12. Molecular identification and transmission studies of X-cell parasites from Atlantic cod Gadus morhua (Gadiformes: Gadidae) and the northern black flounder Pseudopleuronectes obscurus (Pleuronectiformes: Pleuronectidae)

    PubMed Central

    2011-01-01

    Background Epidermal pseudotumours from Hippoglossoides dubius and Acanthogobius flavimanus in Japan and gill lesions in Limanda limanda from the UK have been shown to be caused by phylogenetically related protozoan parasites, known collectively as X-cells. However, the phylogenetic position of the X-cell group is not well supported within any of the existing protozoan phyla and they are currently thought to be members of the Alveolata. Ultrastructural features of X-cells in fish pseudotumours are somewhat limited and no typical environmental stages, such as spores or flagellated cells, have been observed. The life cycles for these parasites have not been demonstrated and it remains unknown how transmission to a new host occurs. In the present study, pseudobranchial pseudotumours from Atlantic cod, Gadus morhua, in Iceland and epidermal pseudotumours from the northern black flounder, Pseudopleuronectes obscurus, in Japan were used in experimental transmission studies to establish whether direct transmission of the parasite is achievable. In addition, X-cells from Atlantic cod were sequenced to confirm whether they are phylogenetically related to other X-cells and epidermal pseudotumours from the northern black flounder were analysed to establish whether the same parasite is responsible for infecting different flatfish species in Japan. Results Phylogenetic analyses of small subunit ribosomal DNA (SSU rDNA) sequence data from Atlantic cod X-cells show that they are a related parasite that occupies a basal position to the clade containing other X-cell parasites. The X-cell parasite causing epidermal pseudotumours in P. obscurus is the same parasite that causes pseudotumours in H. dubius. Direct, fish to fish, transmission of the X-cell parasites used in this study, via oral feeding or injection, was not achieved. Non-amoeboid X-cells are contained within discrete sac-like structures that are loosely attached to epidermal pseudotumours in flatfish; these X-cells are

  13. WINTER FLOUNDER PSUEDOPLEURONECTES AMERICANUS HATCHING SUCCESS AS A FUNCTION OF BURIAL DEPTH IN THE LABORATORY

    EPA Science Inventory

    Previous experiments have shown that viable hatch of winter flounder eggs is reduced when the eggs are buried by as little as one half of one egg diameter (approximately 0.5 mm of sediment). This sensitivity to burial has resulted in seasonal banning of dredging in several north...

  14. Transfer of benzo(a)pyrene from two invertebrate prey species to the winter flounder, Pleuronectes americanus

    SciTech Connect

    Schnitz, A.R.; Taghon, G.L.

    1995-12-31

    Anthropogenic activities in and around the estuarine environment may introduce compounds with the ability to move through trophic levels, resulting in adverse effects to ecosystems and human health. In order to observe the potential for transfer of the carcinogen benzo(a)pyrene (BaP) and its derivatives from prey organisms to a predator, a simplified benthic food chain was constructed. Winter flounder, Pleuronectes americanus, were fed via gavage either softshell clams, Mya arenaria, or sandworms, Neanthes virens, that had previously been exposed to {sup 14}C-benzo(a)pyrene-contaminated sediments and contained 5.05 ng/g and 5.8 ng/g activity, respectively (wet weight). The fish received doses every 48 hours for a total of 192 hours after which time, all tissues except the gall bladder were examined for the presence of BaP using liquid scintillation spectrometry (LSC). The bile was extracted with chloroform/methanol and analyzed via high performance liquid chromatography (HPLC) and LSC. The flounders that were fed worms demonstrated increased metabolism of the PAH relative to those fed clams, as shown by higher concentrations of polar metabolites in the bile. This corresponds with the higher concentration of metabolites recovered in the worm tissue; more than 95% of the recovered activity was in the form of polar derivatives whereas the activity in the clam tissue was almost completely parent benzo(a)pyrene. As was expected, the BaP metabolites formed by the fish were mediated by the differing abilities of the prey species to derivatize the PAH. Thus, macrofaunal interactions with sedimentary-associated polycyclic aromatic hydrocarbons may be important determinants in the fate and transport of these compounds in the benthos.

  15. /sup 32/P-postlabeling analysis of DNA adducts in liver of wild English sole (Parophrys vetulus) and winter flounder (Pseudopleuronectes americanus)

    SciTech Connect

    Varanasi, U.; Reichert, W.L.; Stein, J.E.

    1989-03-01

    The 1-butanol adduct enhancement version of the 32P-postlabeling assay was used to measure the levels of hepatic DNA adducts in the marine flatfish, English sole (Parophrys vetulus), sampled from the Duwamish Waterway and Eagle Harbor, Puget Sound, WA, where they are exposed to high concentrations of sediment-associated chemical contaminants and exhibit an elevated prevalence of hepatic neoplasms. Hepatic DNA was also analyzed from English sole from a reference area (Useless Bay, WA) and from reference English sole treated with organic-solvent extracts of sediments from the two contaminated sites. Autoradiograms of thin-layer chromatograms of 32P-labeled hepatic DNA digests from English sole from the contaminated sites exhibited up to three diagonal radioactive zones, which were not present in autoradiograms of thin-layer chromatogram maps of 32P-labeled DNA digests from English sole from the reference site. These diagonal radioactive zones contained several distinct spots as well as what appeared to be multiple overlapping adduct spots. The levels (nmol of adducts/mol of nucleotides) of total DNA adducts for English sole from Duwamish Waterway and Eagle Harbor were 26 +/- 28 (DS) and 17 +/- 9.6, respectively. All autoradiograms of DNA from fish from the contaminated sites exhibited a diagonal radioactive zone where DNA adducts of chrysene, benzo(a)pyrene, and dibenz(a,h)anthracene, formed in vitro using English sole hepatic microsomes, were shown to chromatograph. English sole treated with extracts of the contaminated sediments had adduct profiles generally similar to those for English sole from the respective contaminated sites.

  16. Rapid amyloid fibril formation by a winter flounder antifreeze protein requires specific interaction with ice.

    PubMed

    Dubé, André; Leggiadro, Cindy; Ewart, Kathryn Vanya

    2016-05-01

    A typically α-helical antifreeze protein (wflAFP-6) from winter flounder, Pseudopleuronectes americanus, forms amyloid fibrils during freezing. In this study, the effects of distinct components of the freezing process were examined. Freezing of wflAFP-6 in the presence of template ice was shown to be necessary for rapid conversion to an amyloid conformation. Neither subfreezing temperature nor phase change was sufficient. Thus, specific interaction with the ice surface was essential. The ice-induced formation of amyloid appeared to be unique to this helical antifreeze, it required high concentrations of protein and it occurred over a range of pH values. These results define a method for rapid formation of amyloid by wflAFP-6 on demand under physiological conditions. PMID:27086686

  17. Carbon and nitrogen isotope ratios of juvenile winter flounder as indicators of inputs to estuarine systems.

    PubMed

    Pruell, Richard J; Taplin, Bryan K

    2015-12-30

    Stable carbon and nitrogen isotope ratios were measured in young-of-the-year (YOY) winter flounder, Pseudopleuronectes americanus, collected from several Rhode Island, USA estuarine systems. These included three coastal lagoons, an estuarine river and Narragansett Bay. The δ(13)C trends observed along transects in several systems showed isotopically depleted terrestrial signals in the upper reaches of the estuaries. Significant differences (P<0.05) in δ(15)N were observed among all estuarine systems and these differences correlated (P<0.01) with human population densities in the watersheds. Although Narragansett Bay has a strong north-south gradient in nutrient concentrations this trend was not reflected in flounder δ(15)N. The northernmost station with the highest nutrient concentrations unexpectedly had significantly lower δ(15)N values. Depleted δ(15)N values at this nutrient-rich station may indicate that concentration-dependent fractionation needs to be considered when using nitrogen isotope ratios in biota to monitor anthropogenic nitrogen inputs in systems with high nitrogen loadings. PMID:26541984

  18. Esophageal desalination of seawater in flounder: role of active sodium transport.

    PubMed

    Parmelee, J T; Renfro, J L

    1983-12-01

    The esophagus of the flounder, Pseudopleuronectes americanus, was studied to determine how salinity of ingested seawater (SW) is decreased before fluid absorption in the intestine. Drinking rate was 2.5 ml X h-1 X kg-1. Stomach fluid osmolality was 45% that of seawater, and intestinal fluid was isosmotic to plasma. Esophagus and stomach were nearly impermeable to 28Mg; thus Mg concentrations were accurate indicators of fluid addition and NaCl removal between pharynx and stomach. Measurements of water and ion fluxes across isolated esophageal epithelium mounted in Ussing chambers and bathed by Ringer solution showed that the tritiated water flux was lower in esophagus than in intestine and that 22Na flux ratio was 1.4 (Jm leads to s/Js leads to m) regardless of acclimation medium (100 or 10% SW). Potential difference was zero, and electrical resistance averaged 90 omega X cm2. Mucosal-to-serosal Na transport was inhibited by 0.1 mM amiloride, 0.1 mM ouabain, and Cl-free medium, whereas 1.0 mM furosemide had no effect. Net esophageal Na absorption (mucosal-to-serosal) averaged 10.0 mumol X h-1 X cm-2 with mucosa exposed to SW and was inhibited 46% by 0.1 mM ouabain. Taken together the above observations suggest a role for both passive and active esophageal Na transport in SW desalination.

  19. Primary structure of insulin and glucagon from the flounder (Platichthys flesus).

    PubMed

    Conlon, J M; Davis, M S; Thim, L

    1987-05-01

    Insulin and glucagon have been isolated from the Brockmann bodies of the flounder, a teleostean fish, and their primary structures established by automated Edman degradation. The A-chain of flounder insulin shows strong homology to the A-chains from the coho salmon (Oncorhynchus kisutch; 100%) and the anglerfish (Lophius americanus; 95%) but homologies in the B-chain region are weaker (salmon 79%, anglerfish 83%). Flounder insulin B-chain contains the novel sequence Val-Val-Pro-Pro at the NH2 terminus and the highly conserved seryl residue at position 10 (B 9 in mammals) is replaced by an alanyl residue. Flounder glucagon is identical to anglerfish glucagon II but shows four amino acid substitutions compared with salmon glucagon. PMID:3556313

  20. Maternal effects and larval survival of marbled sole Pseudopleuronectes yokohamae

    NASA Astrophysics Data System (ADS)

    Higashitani, Tomomi; Takatsu, Tetsuya; Nakaya, Mitsuhiro; Joh, Mikimasa; Takahashi, Toyomi

    2007-07-01

    Maternal effects of animals are the phenotypic influences of age, size, and condition of spawners on the survival and phenotypic traits of offspring. To clarify the maternal effects for marbled sole Pseudopleuronectes yokohamae, we investigated the effects of body size, nutrient condition, and growth history of adult females on egg size, larval size, and starvation tolerance, growth, and feeding ability of offspring. The fecundity of adult females was strongly dependent on body size. Path analysis revealed that the mother's total length positively affected mean egg diameter, meaning that large females spawned large eggs. In contrast, the relative growth rate of adult females negatively affected egg diameter. Egg diameters positively affected both notochord length and yolk sac volume of the larvae at hatching. Under starvation conditions, notochord length at hatching strongly and positively affected days of survival at 14 °C but not at 9 °C. Under adequate food conditions (1000 rotifers L - 1 ), the notochord length of larvae 5 days after hatching positively affected feeding rate, implying that large larvae have high feeding ability. In addition, the mean growth rate of larvae between 0 and 15 days increased with increasing egg diameter under homogenous food conditions, suggesting that larvae hatched from large eggs might have a growth advantage for at least to 15 days after hatching. In marbled sole, these relationships (i.e., mother's body size-egg size-larval size-larval resistance to starvation-larval feeding ability) may help explain recruitment variability.

  1. An Early Warning System for Flounder Disease

    NASA Astrophysics Data System (ADS)

    Xing, Bin; Li, Daoliang; Wang, Jianqin; Duan, Qingling; Wen, Jiwen

    With the constant expansion of the scale and mismanagement in aquaculture,the diseases of flounder occur more and more frequently than before, which has brought great economic losses to fish farmers. For the sake of the problem described above, based on a great number of surveys, the early warning theory of flounder disease, the analysis of the outbreak and development of diseases and the relationship between disease and factors, the logic process of the early warning for flounder disease was confirmed. It consists of five parts: specifying the target, searching for the source, distinguishing the sign, predicting the degree and eliminating the menace. Using the expert survey method the early warning indexes which affect the normal life of the flounder and calculated the range of the water environment factors were also confirmed. Finally, an early warning system was implemented, which can reduce the damage from the flounder disease.

  2. 50 CFR 648.105 - Summer flounder recreational fishing season.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Summer flounder recreational fishing season. 648.105 Section 648.105 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL... Management Measures for the Summer Flounder Fisheries § 648.105 Summer flounder recreational fishing...

  3. 50 CFR 648.105 - Summer flounder recreational fishing season.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Summer flounder recreational fishing season. 648.105 Section 648.105 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL... Management Measures for the Summer Flounder Fisheries § 648.105 Summer flounder recreational fishing...

  4. 50 CFR 648.105 - Summer flounder recreational fishing season.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Summer flounder recreational fishing season. 648.105 Section 648.105 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL... Management Measures for the Summer Flounder Fisheries § 648.105 Summer flounder recreational fishing...

  5. 50 CFR 648.102 - Summer flounder specifications.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ..., recreational landing limits, research set-asides, and other specification measures. The Summer Flounder... to be considered by the Summer Flounder Monitoring Committee are: (1) Research quota set from a range... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Summer flounder specifications....

  6. 50 CFR 648.102 - Summer flounder specifications.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ..., recreational landing limits, research set-asides, and other specification measures. The Summer Flounder... to be considered by the Summer Flounder Monitoring Committee are: (1) Research quota set from a range... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Summer flounder specifications....

  7. 50 CFR 648.104 - Summer flounder minimum fish sizes.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Summer flounder minimum fish sizes. 648... Measures for the Summer Flounder Fisheries § 648.104 Summer flounder minimum fish sizes. (a) Moratorium... minimum sizes in this section apply to whole fish or to any part of a fish found in possession,...

  8. 50 CFR 648.104 - Summer flounder minimum fish sizes.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Summer flounder minimum fish sizes. 648... Measures for the Summer Flounder Fisheries § 648.104 Summer flounder minimum fish sizes. (a) Moratorium... minimum sizes in this section apply to whole fish or to any part of a fish found in possession,...

  9. 50 CFR 648.103 - Summer flounder accountability measures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Summer flounder accountability measures... Management Measures for the Summer Flounder Fisheries § 648.103 Summer flounder accountability measures. (a... subsequent single fishing year recreational sector ACT. (d) Non-landing accountability measures, by...

  10. 50 CFR 648.103 - Summer flounder accountability measures.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Summer flounder accountability measures... Management Measures for the Summer Flounder Fisheries § 648.103 Summer flounder accountability measures. (a... subsequent single fishing year recreational sector ACT. (d) Non-landing accountability measures, by...

  11. Habitat Suitability Index Models: Southern and gulf flounders

    USGS Publications Warehouse

    Enge, Kevin M.; Mulholland, Rosemarie

    1985-01-01

    Both the southern and gulf flounders (paralichthrS lethostigma, f. albi~utta) are important commercial and recreationa species. Catch statlstics for flounder do not differentiate between species; however, southern flounders are more common than gulf flounders except on the gulf coast of Florida (Topp and Hoff 1972). The commercial fishery consists of incidental catches by shrimp trawlers. A total of 853,162 kg (1,880,900 lb) of unclassified flounder worth $572,142 was caught from the Gulf of Mexico in 1976 (U.S. Dep. Commerce 1980). The sport fishery consists of both hook-and-line fishing and gigging.

  12. Antifreeze proteins from the sea raven, Hemitripterus americanus. Further evidence for diversity among fish polypeptide antifreezes

    SciTech Connect

    Slaughter, D.; Fletcher, G.L.; Ananthanarayanan, V.S.; Hew, C.L.

    1981-02-25

    The antifreeze proteins of the sea raven, Hemitripterus americanus, were isolated and compared with other fish antifreeze proteins. The sea raven contains one major protein of molecular weight 14,000-16,000 with little or no carbohydrate. Except for its similar seasonal appearance, the sea raven antifreeze protein differs from other polypeptide antifreeze in its amino acid composition, secondary structure, and immunological specificity. Amino acid analysis of sea raven antifreeze showed that it contains a high amount of half-cystine, hydrophilic amino acids, and only an average amount of alanine. In contrast, all other fish antifreeze proteins contain approximately 60% alanine and no half-cystine residues. Furthermore, the sea raven antifreeze protein is sensitive to sulfhydryl reagents. The antifreeze activity was decreased by 67% in the presence of 0.01 M dithiothreitol. Circular dichroism studies indicated the absence of significant amounts of ..cap alpha..-helix and the possible presence of ..beta..-structure. Antibodies raised against the antifreeze protein did not cross-react with the known polypeptide antifreeze from the winter flounder and shorthorn sculpin (Hew, C.L., Fletcher, G.L., and Ananthanarayanan, V.S. (1980) Can. J. Biochem. 58, 377-383). A specific radioimmunoassay was developed for the sea raven antifreeze protein and was used to quantitate the protein concentration in the fish. The seasonal profile obtained by radioimmunoassay was compatible with the antifreeze activity determined with a freezing point osmometer.

  13. Structural characterization of peptides derived from prosomatostatins I and II isolated from the pancreatic islets of two species of teleostean fish: the daddy sculpin and the flounder.

    PubMed

    Conlon, J M; Davis, M S; Falkmer, S; Thim, L

    1987-11-01

    The primary structures of three peptides from extracts from the pancreatic islets of the daddy sculpin (Cottus scorpius) and three analogous peptides from the islets of the flounder (Platichthys flesus), two species of teleostean fish, have been determined by automated Edman degradation. The structures of the flounder peptides were confirmed by fast-atom bombardment mass spectrometry. The peptides show strong homology to residues (49-60), (63-96) and (98-125) of the predicted sequence of preprosomatostatin II from the anglerfish (Lophius americanus). The amino acid sequences of the peptides suggest that, in the sculpin, prosomatostatin II is cleaved at a dibasic amino acid residue processing site (corresponding to Lys61-Arg62 in anglerfish preprosomatostatin II). The resulting fragments are further cleaved at monobasic residue processing sites (corresponding to Arg48 and Arg97 in anglerfish preprosomatostatin II). In the flounder the same dibasic residue processing site is utilised but cleavage at different monobasic sites takes place (corresponding to Arg50 and Arg97 in anglerfish preprosomatostatin II). A peptide identical to mammalian somatostatin-14 was also isolated from the islets of both species and is presumed to represent a cleavage product of prosomatostatin I. PMID:2889597

  14. Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus: karyotype, allozyme and RAPD analyses

    NASA Astrophysics Data System (ADS)

    You, Feng; Wang, Wei; Xu, Dongdong; Zhu, Xiangping; Ni, Jing; Wu, Zhihao; Xu, Yongli; Wang, Xincheng; Zhang, Peijun

    2009-05-01

    The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes, 12 confirmed hybridization of the paternal and maternal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.

  15. 77 FR 64239 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-19

    ... Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested for the State of New York...), Commerce. ACTION: Temporary rule; closure. SUMMARY: NMFS announces that the 2012 summer flounder commercial... fisheries permit for the summer flounder fishery may not land summer flounder in New York for the...

  16. 78 FR 51096 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-20

    ... Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested for the Commonwealth of... (NOAA), Commerce. ACTION: Temporary rule; closure. SUMMARY: NMFS announces that the 2013 summer flounder... commercial Federal fisheries permit for the summer flounder fishery may not land summer flounder...

  17. 78 FR 70890 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-27

    ... Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested for the State of New Jersey...), Commerce. ACTION: Temporary rule; closure. SUMMARY: NMFS announces that the 2013 summer flounder commercial... fisheries permit for the summer flounder fishery may not land summer flounder in New Jersey for...

  18. 78 FR 72585 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-03

    ... Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested for the Commonwealth of... (NOAA), Commerce. ACTION: Temporary rule; closure. SUMMARY: NMFS announces that the 2013 summer flounder... Federal fisheries ] permit for the summer flounder fishery may not land summer flounder in Virginia...

  19. ELEMENTAL AND ISOTPOIC FINGERPRINTS OF JUVENILE WINTER FLOUNDER (PLEURONECTES AMERICANUS) FROM NARRAGANSETT BAY, RI (USA) AND SURROUNDING COASTAL PONDS

    EPA Science Inventory

    Elemental fingerprinting has become a powerful tool in fisheries science for identifying fish migration patterns, seasonal changes in habitat use, and for delineating the nursery origins of adult fish populations. In this study, we investigated whether elemental and isotopic sign...

  20. Spatial and Temporal Variability of Elemental Signautres in Juvenile Winter Flounder (Psuedopleuronectes americanus): Implications for Natal Connectivity

    EPA Science Inventory

    Elemental signatures in otoliths (fish ear-stones) have become a powerful tool in fisheries science for identifying fish migration patterns, reconstructing environmental histories, and for delineating the nursery origins of adult fish populations. Assessing connectivity between a...

  1. Visual optics in toads (Bufo americanus).

    PubMed

    Mathis, U; Schaeffel, F; Howland, H C

    1988-06-01

    Aspects of visual optics were investigated in the American toad (Bufo americanus). The development of the refractive state of the eye during metamorphosis was followed with IR photoretinoscopy. Frozen sections documented the changes in optical parameters before and after metamorphosis. There is a difference in light sensitivity between juvenile and adult toads. Binocular accommodation in adult toads was observed. 1. IR photoretinoscopic measurements showed that the refractive state of the eye changed very rapidly during metamorphosis, about 10 D/h while the animal entered the terrestrial habitat. 2. Frozen sections showed that the almost spherical lens in a tadpole eye had flattened in a just metamorphosed toad's eye while at the same time the distance of the lens to the retina had decreased. However, the morphological measurements were not sufficiently sensitive to record the relatively small changes in ocular dimensions that were responsible for the rapid changes in refractive state during metamorphosis. 3. Schematic eyes, with homogeneous and non homogeneous lenses, were constructed for tadpoles, juvenile toads, and adult toads. 4. Nonparaxial raytracing studies in schematic eyes suggested that the lenses of animals of the three developmental stages tadpole, juvenile toad, and adult are not homogeneous but have a refractive index gradient. The raytracing studies indicated that the refractive index gradient is different for the different developmental stages, being highest in the tadpole lens. 5. The observations of toads during feeding behavior at different light levels showed an increased light sensitivity in the adult nocturnal toads in contrast to the juvenile animals, which are diurnal. The increased light sensitivity could partly be explained with an increase in aperture and an increase in red rod outer segments. To fully explain the higher light sensitivity in adult toads, changes in neuronal parameters had to be assumed. 6. Retinoscopic measurements of

  2. Eastern equine encephalitis in moose (Alces americanus) in northeastern Vermont.

    PubMed

    Mutebi, John-Paul; Swope, Bethany N; Saxton-Shaw, Kali D; Graham, Alan C; Turmel, Jon P; Berl, Erica

    2012-10-01

    During fall 2010, 21 moose (Alces americanus) sera collected in northeastern Vermont were screened for eastern equine encephalitis virus (EEEV) antibodies using plaque reduction neutralization tests. Six (29%) were antibody positive. This is the first evidence of EEEV activity in Vermont, and the second report of EEEV antibodies in moose.

  3. Sarcocystis oreamni n. sp. from the mountain goat (oreamnos americanus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous species of Sarcocystis have been reported from wild ruminants but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of 3 of 7 mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; 1 had ...

  4. Use of a Marine Vertebrate, the Flounder, in the Physiology Teaching Laboratory

    ERIC Educational Resources Information Center

    Bruce, David S.; Linden, Donald G.

    1973-01-01

    Describes two experiments in physiology using a readily available marine vertebrate, the flounder. Representative results are presented from experiments which measure the effect of excess potassium on the flounder EKG, and the effect of temperature on heart rate of the starry flounder. (JR)

  5. 50 CFR 648.110 - Summer flounder framework adjustments to management measures.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Summer flounder framework adjustments to... UNITED STATES Management Measures for the Summer Flounder Fisheries § 648.110 Summer flounder framework... are otherwise introducing new concepts may require an amendment of the FMP instead of a...

  6. 50 CFR 648.110 - Summer flounder framework adjustments to management measures.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Summer flounder framework adjustments to... UNITED STATES Management Measures for the Summer Flounder Fisheries § 648.110 Summer flounder framework... are otherwise introducing new concepts may require an amendment of the FMP instead of a...

  7. 50 CFR 648.110 - Summer flounder framework adjustments to management measures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Summer flounder framework adjustments to... UNITED STATES Management Measures for the Summer Flounder Fisheries § 648.110 Summer flounder framework... are otherwise introducing new concepts may require an amendment of the FMP instead of a...

  8. 50 CFR 648.110 - Summer flounder framework adjustments to management measures.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 10 2011-10-01 2011-10-01 false Summer flounder framework adjustments to... UNITED STATES Management Measures for the Summer Flounder Fisheries § 648.110 Summer flounder framework... instead of a framework adjustment. (2) MAFMC recommendation. After developing management actions...

  9. Taxonomy of the early life stages of arrowtooth flounder (Atheresthes stomias) and Kamchatka flounder (A. evermanni) in the eastern Bering Sea, with notes on distribution and condition

    NASA Astrophysics Data System (ADS)

    De Forest, Lisa; Duffy-Anderson, J. T.; Heintz, R. A.; Matarese, A. C.; Siddon, E. C.; Smart, T. I.; Spies, I. B.

    2014-11-01

    Arrowtooth flounder (Atheresthes stomias) and Kamchatka flounder (A. evermanni) are closely related flatfish species that co-occur in the eastern Bering Sea. As adults, arrowtooth flounder can be distinguished from Kamchatka flounder; however, larvae and early juveniles can only be indentified to the genus level due to morphological similarities. This has precluded studies of ecology for the early life stages of both species in the eastern Bering Sea. In this study, we developed a genetic technique to identify the larvae and early juveniles of the two species using mtDNA cytochrome oxidase subunit I (COI). Genetically identified specimens were then examined to determine a visual identification method based on pigment patterns and morphology. Specimens 6.0-12.0 mm SL and≥18.0 mm SL can be identified to the species level, but species identification of individuals 12.1-17.9 mm SL by visual means alone remains elusive. The distribution of larvae (<25.0 mm SL) of both arrowtooth flounder and Kamchatka flounder is similar in the eastern Bering Sea; however, juvenile (≥25.0 mm SL) Kamchatka flounder occur closer to the shelf break and in deeper water than juvenile arrowtooth flounder. Condition was determined for larvae and juveniles of each species by analyzing lipid content (%) and energy density (kJ/g dry mass). Kamchatka flounder larvae on average had higher lipid content than arrowtooth flounder larvae, but were also larger on average than arrowtooth flounder larvae in the summer. When corrected for length, both species had similar lipid content in the larval and juvenile stages.

  10. 50 CFR 648.106 - Summer flounder possession restrictions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... flounder are filleted into single (butterfly) fillets, each fillet is deemed to be from one whole summer... trawl vessels issued a permit under § 648.4(a)(3) that fish with or possess nets or pieces of net on... for inspection in standard 100-lb (45.3-kg) totes or fish boxes having a liquid capacity of 18.2...

  11. 50 CFR 648.106 - Summer flounder possession restrictions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... flounder are filleted into single (butterfly) fillets, each fillet is deemed to be from one whole summer... trawl vessels issued a permit under § 648.4(a)(3) that fish with or possess nets or pieces of net on... for inspection in standard 100-lb (45.3-kg) totes or fish boxes having a liquid capacity of 18.2...

  12. 50 CFR 648.106 - Summer flounder possession restrictions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... flounder are filleted into single (butterfly) fillets, each fillet is deemed to be from one whole summer... trawl vessels issued a permit under § 648.4(a)(3) that fish with or possess nets or pieces of net on... for inspection in standard 100-lb (45.3-kg) totes or fish boxes having a liquid capacity of 18.2...

  13. 50 CFR Figure 10 to Part 223 - Flounder TED

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 10 2012-10-01 2012-10-01 false Flounder TED 10 Figure 10 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig. 10 Figure 10...

  14. 50 CFR Figure 11 to Part 223 - Modified Flounder TED

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 10 2012-10-01 2012-10-01 false Modified Flounder TED 11 Figure 11 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig....

  15. 50 CFR Figure 10 to Part 223 - Flounder TED

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 9 2011-10-01 2011-10-01 false Flounder TED 10 Figure 10 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig. 10 Figure 10...

  16. 50 CFR Figure 10 to Part 223 - Flounder TED

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 10 2014-10-01 2014-10-01 false Flounder TED 10 Figure 10 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig. 10 Figure 10...

  17. 50 CFR Figure 11 to Part 223 - Modified Flounder TED

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 10 2014-10-01 2014-10-01 false Modified Flounder TED 11 Figure 11 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig....

  18. 50 CFR Figure 11 to Part 223 - Modified Flounder TED

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 10 2013-10-01 2013-10-01 false Modified Flounder TED 11 Figure 11 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig....

  19. 50 CFR Figure 10 to Part 223 - Flounder TED

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 7 2010-10-01 2010-10-01 false Flounder TED 10 Figure 10 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig. 10 Figure 10...

  20. 50 CFR Figure 10 to Part 223 - Flounder TED

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 10 2013-10-01 2013-10-01 false Flounder TED 10 Figure 10 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES Pt. 223, Fig. 10 Figure 10...

  1. Properties of proteolytic toxin of Vibrio anguilolarum from diseased flounder

    NASA Astrophysics Data System (ADS)

    Mo, Zhao-Lan; Chen, Shi-Yong; Zhang, Pei-Jun

    2002-12-01

    Extracellular products (ECP) produced by Vibrio anguillarum strain M3 originally isolated from diseased flounder ( Paralichthys olivaceus) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity against azocasin was detected. Bacterium M2 showed highest growth and protease activity at 25°C. The protease present in ECP showed maximal activity at pH 8 and 55°C; was completely inactivated by application of 80°C heat for 30 min; was completely inhibited by EDTA and HgCl2, and was partially inhibited by PMSF, SDS, MnCl2 and iodoacetic acid; but not inhibited by CaCl2 and MgCl2. The ECP was toxic to flounder fish at LD50 values of 3.1 μg protein/g body weight. The addition of HgCl2 and application of heat at 50°C decreased the lethal toxicity of ECP. When heated at 100°C, ECP lethality to flounder was completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerious lymphocytes in the liver. These results showed that ECP protease was a lethal factor produced by the bacterium V. anguillarum M3.

  2. Cretinism in a North American black bear (Ursus americanus).

    PubMed

    Duncan, Robert B; Jones, Jeryl C; Moll, H David; Moon, Martha M; Blodgett, Dennis J; Vaughan, Michael R

    2002-01-01

    Congenital hyperplastic goiter and cretinism were documented in a 16 month-old male North American black bear (Ursus americanus). The cub was captured at approximately 8 months of age and maintained for an additional 8 months in captivity. Clinical signs included growth retardation, clumsiness, and facial dysmorphism. Hypothyroidism was documented by determining serum triiodothyronine (T3) and thyroxine (T4) levels. Lysosomal storage disease was ruled out by measuring various lysosomal enzyme activities. Serologic, radiographic, computed tomographic, necropsy, and histopathologic findings were consistent with congenital hypothyroidism and cretinism. PMID:11866041

  3. Pilot attempt to grow winter flounders commercially. Completion report 15 Feb 77-30 Jun 80

    SciTech Connect

    Sawyer, P.J.; Hoornbeek, F.K.

    1980-09-01

    The first part of this report contains the details and results of laboratory efforts to raise flounders through the early developmental stages including metamorphosis. The second part reports the efforts to grow out newly metamorphosed flounders to commercial size. In order to produce marketable sized winter flounders in heated water, it will be necessary to assure availability of brood stock and an abundance of live food both prior and subsequent to metamorphosis.

  4. Hookworm (Necator americanus) larval enzymes disrupt human vascular endothelium.

    PubMed

    Souadkia, Nahed; Brown, Alan; Leach, Lopa; Pritchard, David I

    2010-09-01

    Knowledge of the molecular mechanisms used by Necator americanus larvae to penetrate the human skin and the vasculature would aid the development of effective vaccines against this important pathogen. In this work, the impact of N. americanus exsheathing fluid (EF) and excretory/secretory products (ES) on the endothelial barrier was examined using human umbilical vein endothelial cells (HUVEC). Cellular responses were assessed by investigating molecular changes at cell-cell junctions and by determining levels of secreted IL-6, IL-8, and vascular endothelial growth factor (VEGF) in the culture medium. It would appear that a repertoire of larval proteases caused a dose-related increase in endothelial permeability as characterized by a decrease in monolayer resistance with increased permeation of tracer-albumin. These barrier changes were associated with disruption of junctional vascular endothelial cadherin (VE-cadherin) and F-actin and an increase in endothelial secretion of IL-6 and IL-8. Our data suggest that larval proteases play an important role in negotiating the endothelium.

  5. Ontogeny of the lymphoid organs of Japanese flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Liu, Yun; Jiang, Guoliang; Zhang, Shicui

    2004-10-01

    Histological study on the ontogeny of the lymphoid organs, kidney, thymus and spleen of Japanese flounder, Paralichthys olivaceus, from hatching to 40 d was carried out. The pronephric kidney duct appeared early in hatching although the primordial haemopoietic stem cells were observed within a week after hatching. The spleen was first seen after 8d of hatching. The thymus appeared after 15d, situated near the pronephric kidney. Small lymphoid cells appeared during the later phase of the post-larval stage in the sequence of thymus, kidney and spleen. During the 40d of observations, there were no distinct inner or outer zones in thymus and no red or white pulp in spleen. These results suggest that the nonspecific defense immune system plays a very important role in the early larval stage of Japanese flounder.

  6. 75 FR 70192 - Fisheries of the Northeastern United States; Summer Flounder, Scup, and Black Sea Bass Fisheries...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-17

    ... United States; Summer Flounder, Scup, and Black Sea Bass Fisheries; 2011 Summer Flounder, Scup, and Black..., and black sea bass fisheries and provides notice of three projects that may be requesting Exempted...-Aside (RSA) program. The implementing regulations for the Summer Flounder, Scup, and Black Sea...

  7. Transcriptional Responses of Olive Flounder (Paralichthys olivaceus) to Low Temperature

    PubMed Central

    Hu, Jinwei; You, Feng; Wang, Qian; Weng, Shenda; Liu, Hui; Wang, Lijuan; Zhang, Pei-Jun; Tan, Xungang

    2014-01-01

    The olive flounder (Paralichthys olivaceus) is an economically important flatfish in marine aquaculture with a broad thermal tolerance ranging from 14 to 23°C. Cold-tolerant flounder that can survive during the winter season at a temperature of less than 14°C might facilitate the understanding of the mechanisms underlying the response to cold stress. In this study, the transcriptional response of flounder to cold stress (0.7±0.05°C) was characterized using RNA sequencing. Transcriptome sequencing was performed using the Illumina MiSeq platform for the cold-tolerant (CT) group, which survived under the cold stress; the cold-sensitive (CS) group, which could barely survive at the low temperature; and control group, which was not subjected to cold treatment. In all, 29,021 unigenes were generated. Compared with the unigene expression profile of the control group, 410 unigenes were up-regulated and 255 unigenes were down-regulated in the CT group, whereas 593 unigenes were up-regulated and 289 unigenes were down-regulated in the CS group. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that signal transduction, lipid metabolism, digestive system, and signaling molecules and interaction were the most highly enriched pathways for the genes that were differentially expressed under cold stress. All these pathways could be assigned to the following four biological functions for flounder that can survive under cold stress: signal response to cold stress, cell repair/regeneration, energy production, and cell membrane construction and fluidity. PMID:25279944

  8. 77 FR 52262 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-29

    ... quota for summer flounder for the 2012 fishing year is 13,136,001 lb (5,958,490 kg) (76 FR 82189... Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested for the Commonwealth of... (NOAA), Commerce. ACTION: Temporary rule; closure. SUMMARY: NMFS announces that the 2012 summer...

  9. 77 FR 74390 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Available...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-14

    ... summer flounder for the 2012 fishing year is 13,136,001 lb (5,958,490 kg) (76 FR 82189, December 30, 2011... Northeastern United States; Summer Flounder Fishery; Commercial Quota Available for the State of New York To... Administration (NOAA), Commerce. ACTION: Temporary rule. SUMMARY: NMFS announces that the 2012 summer...

  10. OCCURRENCE AND ORIENTATION OF PARALICHTHID FLOUNDERS (BOTHIDAE: PARALICHTYS) ON AN INTERTIDAL BEACH

    EPA Science Inventory

    Middaugh, Douglas P. and Charles L. McKenney, Jr. 2003. Occurrence and Orientation of Flounders (Bothidae: Paralichthys) on an Intertidal Beach. J. North Carol. Acad. Sci. 119(4):157-171. (ERL,GB 1172).

    The intertidal movement and burying pattern of paralichthid flounders...

  11. 77 FR 75569 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-21

    ... summer flounder for the 2012 fishing year is 13,136,001 lb (5,958,490 kg) (76 FR 82189, December 30, 2011... advise New Jersey that the quota has been harvested and to advise vessel permit holders and dealer permit holders that no Federal commercial quota is available for landing summer flounder in New Jersey....

  12. Habitat selection of stone and starry flounders in an estuary in relation to feeding and survival

    NASA Astrophysics Data System (ADS)

    Tomiyama, Takeshi; Omori, Michio

    2008-09-01

    Juveniles of both stone flounder Platichthys bicoloratus and starry flounder Platichthys stellatus utilize estuaries as nursery grounds. To understand their habitat selection and the functions of habitats such as food supply, we defined the seasonal distribution of recently settled fish of these species in shallow nursery areas and investigated their feeding habits in the Natori River estuary, Japan. Distribution of stone flounder was limited to the lower estuary (<3 km upriver from the mouth) and stone flounder were most abundant near the mouth. Recently settled starry flounder were first detected further upstream in areas characterized by low salinity <10 and by the absence of the predatory sand shrimp Crangon uritai. Early juvenile stone and starry flounders consumed mainly siphons of the bivalve Nuttallia olivacea and the mysid Neomysis awatschensis, respectively; however, 1- and 2-yr-old fish of both stone and starry flounders fed mainly on the bivalve siphons. These results indicate that habitat selections of juvenile stone and starry flounders enable utilization of preferred prey and predator avoidance, respectively, and that non-overlap of these species' habitats results in avoidance of inter-specific competition for food between these two species.

  13. HYDROCEPHALUS IN THREE JUVENILE NORTH AMERICAN BLACK BEARS (URSUS AMERICANUS).

    PubMed

    Ferguson, Sylvia H; Novak, Janelle; Hecht, Silke; Craig, Linden E

    2016-06-01

    Hydrocephalus has been reported in a variety of species, including the North American black bear ( Ursus americanus ). This report describes three cases of hydrocephalus in this species from wild bears aged 3-4 mo considered retrospectively from necropsy records of one institution. Clinical signs included cortical blindness and ataxia. Primary gross findings were doming of the skull, gyri compression and flattening, and lateral ventricle dilation. Two cases had severe bilateral ventricular dilation with loss of the septum pellucidum; atrophy of the surrounding corpus callosum; and bilateral periventricular tears involving the caudate nuclei, internal capsule, and adjacent cerebrum. Histologically, the cases with periventricular tearing had severe axonal loss and degeneration, malacia, hemorrhage, and variable periventricular astrocytosis. All cases were likely congenital, given the bears' age and lack of an apparent acquired obstruction. PMID:27468040

  14. Tetranucleotide microsatellite loci from the black bear (Ursus americanus)

    USGS Publications Warehouse

    Sanderlin, J.S.; Faircloth, B.C.; Shamblin, B.; Conroy, M.J.

    2009-01-01

    We describe primers and polymerase chain reaction conditions to amplify 21 tetranucleotide microsatellite DNA loci in black bears (Ursus americanus). We tested primers using individuals from two populations, one each in Georgia and Florida. Among individuals from Georgia (n = 29), primer pairs yielded an average of 2.9 alleles (range, one to four) and an average observed heterozygosity (HO) of 0.50 (range, 0.00 to 0.79). Among individuals from Florida (n = 19), primer pairs yielded an average of 5.7 alleles (range, one to 14) and an HO of 0.55 (range, 0.00 to 1.00). A comparison of previously developed markers with individuals from Georgia suggests that bear populations in Georgia and Florida have reduced allelic diversity relative to other populations. ?? 2008 The Authors.

  15. Induction of pseudopregnancy in the American Black Bear (Ursus americanus).

    PubMed

    Schulz, Laura Clamon; Nelson, Ralph A; Pyter, Leah M; Bahr, Janice M

    2003-08-01

    The aim of this study was to determine whether American Black Bears (Ursus americanus) can experience a pseudopregnancy of the same duration as pregnancy. To do this, we treated three nonmated, captive female bears with human chorionic gonadotropin (hCG) during one breeding season, and saline during another. Progesterone concentrations were measured in monthly blood samples to determine whether pseudopregnancy had occurred. Elevated progesterone concentrations were observed in two out of three bears treated with hCG. We conclude that 1) Elevated progesterone concentrations can be induced in black bears by injection of 35 U/kg hCG during the mating season. 2) Bears can experience a pseudopregnancy, identical in length to pregnancy, in which progesterone profiles are indistinguishable from those of pregnancy.

  16. Induction of pseudopregnancy in the American Black Bear (Ursus americanus).

    PubMed

    Schulz, Laura Clamon; Nelson, Ralph A; Pyter, Leah M; Bahr, Janice M

    2003-08-01

    The aim of this study was to determine whether American Black Bears (Ursus americanus) can experience a pseudopregnancy of the same duration as pregnancy. To do this, we treated three nonmated, captive female bears with human chorionic gonadotropin (hCG) during one breeding season, and saline during another. Progesterone concentrations were measured in monthly blood samples to determine whether pseudopregnancy had occurred. Elevated progesterone concentrations were observed in two out of three bears treated with hCG. We conclude that 1) Elevated progesterone concentrations can be induced in black bears by injection of 35 U/kg hCG during the mating season. 2) Bears can experience a pseudopregnancy, identical in length to pregnancy, in which progesterone profiles are indistinguishable from those of pregnancy. PMID:12884278

  17. HYDROCEPHALUS IN THREE JUVENILE NORTH AMERICAN BLACK BEARS (URSUS AMERICANUS).

    PubMed

    Ferguson, Sylvia H; Novak, Janelle; Hecht, Silke; Craig, Linden E

    2016-06-01

    Hydrocephalus has been reported in a variety of species, including the North American black bear ( Ursus americanus ). This report describes three cases of hydrocephalus in this species from wild bears aged 3-4 mo considered retrospectively from necropsy records of one institution. Clinical signs included cortical blindness and ataxia. Primary gross findings were doming of the skull, gyri compression and flattening, and lateral ventricle dilation. Two cases had severe bilateral ventricular dilation with loss of the septum pellucidum; atrophy of the surrounding corpus callosum; and bilateral periventricular tears involving the caudate nuclei, internal capsule, and adjacent cerebrum. Histologically, the cases with periventricular tearing had severe axonal loss and degeneration, malacia, hemorrhage, and variable periventricular astrocytosis. All cases were likely congenital, given the bears' age and lack of an apparent acquired obstruction.

  18. Multi-decadal variation in size of juvenile Summer Flounder (Paralichthys dentatus) in Chesapeake Bay

    NASA Astrophysics Data System (ADS)

    Nys, Lauren N.; Fabrizio, Mary C.; Tuckey, Troy D.

    2016-01-01

    During the last quarter-century, management of Summer Flounder Paralichthys dentatus along the Atlantic coast resulted in significant increases in abundance such that rebuilding targets were recently achieved. Although spawning stock biomass is high, recruitment of young-of-the-year (YOY) Summer Flounder remains variable. Chesapeake Bay is one of the principal nursery areas for this species, but processes such as growth and survival that affect production of YOY Summer Flounder in this estuary have not been explored. Here, we investigated the relationship between abundance and size of Summer Flounder recruits from the 1988 to 2012 year classes in Chesapeake Bay. We also considered the effects of environmental factors on fish size because conditions in the bay vary spatially during the time that fish occupy nursery areas. To describe variations in Summer Flounder size, we used monthly length observations from 13,018 YOY fish captured by bottom trawl from the lower Chesapeake Bay and the James, York, and Rappahannock river subestuaries where Summer Flounder are commonly observed. We applied a generalized additive model to describe spatial, temporal, and environmental effects on observed fish size; we also considered the density of Summer Flounder and an index of productivity as factors in the model. Summer Flounder in Chesapeake Bay exhibited density-dependent and spatially related variations in mean length: larger fish were found mostly in the Bay and smaller fish in the subestuaries. Additionally, low (< 13 °C) and high (> 26 °C) temperatures and low salinities (< 10 psu) had a negative effect on fish size, indicating that individuals found in these environments were typically smaller than conspecifics inhabiting areas of moderate temperatures and higher salinities. Variable nursery habitat conditions in temperate estuaries affect fish size and, subsequently, may influence production of Summer Flounder year classes through effects on maturation and survival. As

  19. Multi-decadal variation in size of juvenile Summer Flounder (Paralichthys dentatus) in Chesapeake Bay

    NASA Astrophysics Data System (ADS)

    Nys, Lauren N.; Fabrizio, Mary C.; Tuckey, Troy D.

    2015-09-01

    During the last quarter-century, management of Summer Flounder Paralichthys dentatus along the Atlantic coast resulted in significant increases in abundance such that rebuilding targets were recently achieved. Although spawning stock biomass is high, recruitment of young-of-the-year (YOY) Summer Flounder remains variable. Chesapeake Bay is one of the principal nursery areas for this species, but processes such as growth and survival that affect production of YOY Summer Flounder in this estuary have not been explored. Here, we investigated the relationship between abundance and size of Summer Flounder recruits from the 1988 to 2012 year classes in Chesapeake Bay. We also considered the effects of environmental factors on fish size because conditions in the bay vary spatially during the time that fish occupy nursery areas. To describe variations in Summer Flounder size, we used monthly length observations from 13,018 YOY fish captured by bottom trawl from the lower Chesapeake Bay and the James, York, and Rappahannock river subestuaries where Summer Flounder are commonly observed. We applied a generalized additive model to describe spatial, temporal, and environmental effects on observed fish size; we also considered the density of Summer Flounder and an index of productivity as factors in the model. Summer Flounder in Chesapeake Bay exhibited density-dependent and spatially related variations in mean length: larger fish were found mostly in the Bay and smaller fish in the subestuaries. Additionally, low (< 13 °C) and high (> 26 °C) temperatures and low salinities (< 10 psu) had a negative effect on fish size, indicating that individuals found in these environments were typically smaller than conspecifics inhabiting areas of moderate temperatures and higher salinities. Variable nursery habitat conditions in temperate estuaries affect fish size and, subsequently, may influence production of Summer Flounder year classes through effects on maturation and survival. As

  20. Molecular immune response of the American lobster (Homarus americanus) to the White Spot Syndrome Virus.

    PubMed

    Clark, K Fraser; Greenwood, Spencer J; Acorn, Adam R; Byrne, Philip J

    2013-11-01

    The adult American lobster (Homarus americanus) is susceptible to few naturally occurring pathogens, and no viral pathogen is known to exist. Despite this, relatively little is known about the H. americanus immune system and nothing is known about its potential viral immune response. Hundreds of rural communities in Atlantic Canada rely on the lobster fishery for their economic sustainability and could be devastated by large-scale pathogen-mediated mortality events. The White Spot Syndrome Virus is the most economically devastating viral pathogen to global shrimp aquaculture production and has been proposed to be capable of infecting all decapod crustaceans including the European Lobster. An in vivo WSSV injection challenge was conducted in H. americanus and WSSV was found to be capable of infecting and replicating within lobsters held at 20°C. The in vivo WSSV challenge also generated the first viral disease model of H. americanus and allowed for the high-throughput examination of transcriptomic changes that occur during viral infection. Microarray analysis found 136 differentially expressed genes and the expression of a subset of these genes was verified using RT-qPCR. Anti-lipopolysaccharide isoforms and acute phase serum amyloid protein A expression did not change during WSSV infection, contrary to previous findings during bacterial and parasitic infection of H. americanus. This, along with the differential gene expression of thioredoxin and trypsin isoforms, provides compelling evidence that H. americanus is capable of mounting an immune response specific to infection by different pathogen classes.

  1. Occurrence, distribution and prey items of juvenile marbled sole Pseudopleuronectes yokohamae around a submarine groundwater seepage on a tidal flat in southwestern Japan

    NASA Astrophysics Data System (ADS)

    Hata, Masaki; Sugimoto, Ryo; Hori, Masakazu; Tomiyama, Takeshi; Shoji, Jun

    2016-05-01

    Occurrence, distribution and prey items of juvenile marbled sole Pseudopleuronectes yokohamae were investigated around a submarine groundwater seepage on a tidal flat in southwestern Japan. Spatial distribution of radon-222 (222Rn) concentration in water showed more submarine groundwater seepage in the offshore area. The lower salinities at offshore sampling stations corresponded with the highest 222Rn concentrations. Juvenile marbled sole were collected from March through June with seasonal peak in April in 2013 and 2014. Mean abundance of juvenile marbled sole was highest at the second most offshore station where high submarine groundwater seepage was indicated. Major prey items in the stomachs of the marbled sole at the post-settlement stage (10-40 mm) were small crustaceans such as cumaceans and gammarids, which were partially replaced with polychaetes in larger juveniles (40-50 mm). Abundance of these major prey items was also higher at offshore stations. A negative correlation between gammarid abundance and salinity indicated a higher concentration of gammarids around the area of high submarine groundwater seepage, a pattern not observed for the other major prey organisms. Stable isotope analysis showed greater dependence of post-settlement stage marbled sole on the small crustaceans with low δ13C indicating that nutrients of terrestrial origin contribute to production of the juvenile marbled sole on the tidal flat.

  2. Early growth and development of reciprocal hybrids of the starry flounder Platichthys stellatus and stone flounder Kareius bicoloratus.

    PubMed

    Yamashita, Y T; Aritaki, M; Kurita, Y; Tanaka, M

    2014-05-01

    Larval growth and development of hybrid flounder were observed and compared with those of their parent species. The reciprocal hybrids of female starry flounder Platichthys stellatus and male stone flounder Kareius bicoloratus (hybrid Sb) and of female K. bicoloratus and male P. stellatus (hybrid Bs) both survived and grew to juveniles. Development was divided into nine stages (A-I). Many of the hybrids' traits were identical and intermediate to those of their parents. The position of the eye, however, was primarily sinistral in both hybrids (80% in Sb and 76% in Bs), a trait possessed by P. stellatus (80%) in the western Pacific Ocean. The daily growth rates of the larvae were similar. In the parent species, development was more rapid in P. stellatus than in K. bicoloratus whereas rate of development was intermediate in both Sb and Bs hybrids. The size at settlement [standard length (LS ) at stage H (mean ± s.d.)] was 9·82 ± 1·47 mm for the hybrid Sb and 9·99 ± 0·90 mm for the hybrid Bs, while the minimum age at metamorphosis (initial age at stage H) was 29 days after hatching (DAH) in both hybrids. In comparison, LS at settlement in parent species was 6·43 ± 0·25 mm for P. stellatus and 12·87 ± 1·29 mm for K. bicoloratus. Minimum age at metamorphosis for the parents was 23 DAH at stage G in P. stellatus and 34 DAH at stage H in K. bicoloratus. Thus, the timing of settlement of hybrids was different from that of their parent species. These traits may occur with high frequency in a natural habitat.

  3. Identification and expression analysis of hepcidin-like antimicrobial peptides in bony fish.

    PubMed

    Douglas, Susan E; Gallant, Jeffrey W; Liebscher, Ryan S; Dacanay, Andrew; Tsoi, Stephen C M

    2003-01-01

    Antimicrobial peptides play a crucial role as the first line of defense against invading pathogens. Several types of antimicrobial peptides have been isolated from fish, mostly of the cationic alpha-helical variety. Here, we present the cDNA sequences of five highly disulphide-bonded hepcidin-like peptides from winter flounder, Pseudopleuronectes americanus (Walbaum) and two from Atlantic salmon, Salmo salar (L.). These hepcidin-like molecules consist of a 24 amino acid signal peptide and an acidic propiece of 38-40 amino acids in addition to the mature processed peptide of 19-27 amino acids. Exhaustive data mining of GenBank with these sequences revealed that similar peptides are encoded in the genomes of Japanese flounder, rainbow trout, hybrid striped bass and medaka, indicating that they are widespread among fish. Southern hybridization analysis suggests that closely related hepcidin-like genes are present in other flatfish species, and that they exist as a multigene family clustered on the winter flounder genome. Hepcidin variants are differentially expressed during bacterial challenge, during larval development of P. americanus and in different tissues of adult fish.

  4. Subcuticular urate accumulation in an American lobster (Homarus americanus).

    PubMed

    Battison, A L

    2013-05-01

    An unusually "lumpy" lobster, Homarus americanus, was presented to the Atlantic Veterinary College Lobster Science Centre for evaluation. The lobster was weak with numerous pale, raised, and flat areas (diameter, 3-15 mm) on the exoskeleton, some of which were ulcerated. On postmortem examination, the pale areas corresponded to accumulations of viscous to free-flowing white material, which was found in only the subcuticular connective tissues. No internal organs were affected. Direct light examination of nonstained impression smears of the material showed abundant crystals resembling uric acid, amorphous urates, and sodium urate, which were readily soluble in 1 M potassium hydroxide. Wright-Giemsa stained imprints showed numerous fine, rounded, nonstaining granules free in the background and within individual round cells. Fourier-transformed infrared spectroscopy confirmed the presence of urates or mixed urate salts. Hemolymph plasma urea (1.7 mmol/liter) and uric acid (287 μmol/liter) concentrations were slightly higher than those seen with 36-hour emersion. Histologic sections showed aggregates of vacuolated mononuclear cells in the loose subcuticular connective tissue occasionally infiltrating between underlying muscle fibers. Grossly visible urate deposits are occasionally documented in land crabs and rarely reported in the blue crab; none, however, are associated with deformation of the cuticle. Possible etiologies include increased uric acid intake or production or decreased excretion. Anecdotal reports of similarly affected lobsters have been received but are intermittent and undocumented.

  5. Spatial memory in captive American black bears (Ursus americanus).

    PubMed

    Zamisch, Valeria; Vonk, Jennifer

    2012-11-01

    The spatial memory and foraging strategies of four adult captive-born American black bears (Ursus americanus) were explored in four experiments using a simulated foraging task. In the first three experiments, each session consisted of two phases separated by a delay: During the exploration phase, subjects foraged among a set of baited and unbaited sites. During the delay, the same locations were rebaited and subjects were released again and allowed to search the sites (search phase). In Experiments 1a and 1b, different sites were baited each day and the interval between exploration and search was short (4 hr or 15 min). Subjects were not accurate at recovering the food items in either experiment. In Experiment 2, an "informed forager" paradigm was used in which one subject was given privileged knowledge about the location of the food during the exploration phase and was later released with an "uninformed" competitor during the search phase. The bears did not achieve above-chance recovery accuracy even in the presence of a competitor. In Experiment 3, the same two of four sites were continually baited and the bears were released simultaneously over a period of 20 days, with each baiting separated by 2 or 3 days. As a group, the bears' foraging accuracy with repeated baiting and longer intervals approached greater than chance accuracy. Results suggest some limitations on bears' use of spatial memory in captive environments, but reveal the potential for use of spatial memory over longer delays. PMID:22545764

  6. Ontogenetic phase shifts in metabolism in a flounder Paralichthys olivaceus

    PubMed Central

    Yagi, Mitsuharu; Oikawa, Shin

    2014-01-01

    Size-scaling metabolism is widely considered to be of significant importance in biology and ecology. Thus, allometric relationships between metabolic rate () and body mass (M), , have long been a topic of interest and speculation. It has been proposed that intraspecifically metabolic rate scales isometrically or near isometrically with body mass during the early life history in fishes, invertebrates, birds and mammals. We developed a new perspective on intraspecific size-scaling metabolism through determination of metabolic rate in the Japanese flounder, Paralichthys olivaceus, during their early life stages spanning approximately four orders of magnitude in body mass. With the increase of body mass, the Japanese flounder had four distinct negative allometric phases in which three stepwise increases in scaling constants (ai, i = 1–4), i.e. ontogenetic phase shifts in metabolism, occurred with growth during its early life stages at around 0.002, 0.01 and 0.2 g, maintaining each scaling exponent constant in each phase (b = 0.831). These shifts in metabolism during the early life stages are similar to the tiger puffer, Takifugu rubripes. Our results indicate that ontogenetic phase shifts in metabolism are key to understanding intraspecific size-scaling metabolism in fishes. PMID:25412451

  7. The complete genome sequence of 'Candidatus Liberibacter americanus', associated with Citrus huanglongbing.

    PubMed

    Wulff, Nelson A; Zhang, Shujian; Setubal, João C; Almeida, Nalvo F; Martins, Elaine C; Harakava, Ricardo; Kumar, Dibyendu; Rangel, Luiz Thiberio; Foissac, Xavier; Bové, Joseph M; Gabriel, Dean W

    2014-02-01

    Liberibacter spp. form a Rhizobiaceae clade of phloem-limited pathogens of limited host range. Two obligately parasitic species have been sequenced: 'Candidatus Liberibacter asiaticus', which causes citrus huanglongbing (HLB) worldwide, and 'Ca. L. solanacearum', which causes potato "zebra chip" disease. A third (proposed) species, Liberibacter crescens, was isolated from mountain papaya, grown in axenic culture, and sequenced. In an effort to identify common host determinants, the complete genomic DNA sequence of a second HLB species, 'Ca. L. americanus' strain 'São Paulo' was determined. The circular genome of 1,195,201 bp had an average 31.12% GC content and 983 predicted protein encoding genes, 800 (81.4%) of which had a predicted function. There were 658 genes common to all sequenced Liberibacter spp. and only 8 genes common to 'Ca. L. americanus' and 'Ca. L. asiaticus' but not found in 'Ca. L. solanacearum'. Surprisingly, most of the lipopolysaccharide biosynthetic genes were missing from the 'Ca. L. americanus' genome, as well as OmpA and a key regulator of flagellin, all indicating a 'Ca. L. americanus' strategy of avoiding production of major pathogen-associated molecular patterns present in 'Ca. L. asiaticus' and 'Ca. L. solanacearum'. As with 'Ca. L. asiaticus', one of two 'Ca. L. americanus' prophages replicated as an excision plasmid and carried potential lysogenic conversion genes that appeared fragmentary or degenerated in 'Ca. L. solanacearum'.

  8. Direct and indirect effects of southern flounder predation on a spot population: Experimental and model analyses

    SciTech Connect

    Crowder, L.B.; Wright, R.A.; Martin, T.H.; Rice, J.A. . Dept. of Zoology); Rose, K.A. )

    1992-01-01

    We have previously shown that southern flounder (Paralichthys lethostigma Jordan and Gilbert) influence the survival and size-distribution of spot (Leiostomus xanthurus Lafayette) and other small estuarine fishes in experimental ponds. In this paper, we seek to determine whether these of effects can be accounted for by direct size-dependent predation or if there is also evidence for indirect behavioral effects on spot foraging which might alter their survival or population size structure. In our experiment, spot were allowed to grow in the presence and absence of southern flounder in an experimental estuarine pond for 101 days. Each treatment was replicated three times. We also apply a recently published simulation model of the flounder-spot interaction to this experiment to independently test the model and to estimate the direct effects of flounder predation on spot survival and size structure.

  9. Inactivation of Kudoa septempunctata in olive flounder meat by liquid freezing.

    PubMed

    Ohnishi, Takahiro; Akuzawa, Sayuri; Furusawa, Hiroko; Yoshinari, Tomoya; Kamata, Yoichi; Sugita-Konishi, Yoshiko

    2014-01-01

    Kudoa septempunctata in olive flounder meat was inactivated using 3 distinct freezing methods:liquid freezing for 5 min, air blast freezing at -30℃ for 5 h, and -80℃ for 1 h. The fracture curve of olive flounder meat subjected to liquid freezing resembled that of meat stored at 4℃, indicating that the structure of olive flounder muscle was well preserved. In contrast, air blast freezing induced the disappearance of the fracture point in the fracture curve, indicating that there was deterioration in the meat quality. Liquid freezing preserved the transparency of olive flounder meat to the same degree as that of meat stored at 4°C. However, air blast freezing induced meat cloudiness. These results indicate that liquid freezing can be used for K. septempunctata inactivation without affecting the meat quality.

  10. Mercury accumulation and the mercury-PCB-sex interaction in summer flounder

    USGS Publications Warehouse

    Madenjian, Charles P.; Jensen, Olaf P.; Krabbenhoft, David P.; DeWild, John F.; Ogorek, Jacob M.; Vastano, Anthony R.

    2016-01-01

    Patterns in the relative differences in contaminant concentrations between the sexes of mature fish may reveal important behavioral and physiological differences between the sexes. We determined whole-fish total mercury (Hg) concentrations in 23 female summer flounder (Paralichthys dentatus) and 27 male summer flounder from New Jersey coastal waters. To estimate the change in Hg concentration due to release of eggs at spawning, Hg concentration in the somatic tissue and ovaries of 5 of the 23 female summer flounder were also determined. To ascertain whether most of the Hg in the summer flounder was methylmercury (MeHg), whole-fish MeHg concentrations were determined in all 50 summer flounder. Whole-fish Hg concentrations averaged 113 ng/g for females and 111 ng/g for males. Thus, females were 2% higher in Hg concentration than males, on average, but the difference was not statistically significant. Based on Hg determinations in the somatic tissue and ovaries, we predicted that Hg concentration of females would increase by 3.6%, on average, immediately after spawning due to release of eggs. On average, 92% of the Hg in the summer flounder was MeHg. To determine whether the effect of sex on Hg concentration was significantly different from the effect of sex on polychlorinated biphenyl (PCB) concentration, we paired our Hg determinations with PCB determinations from a previous study, and applied regression analysis. Sex significantly interacted with contaminant type (Hg or PCBs), as males were 43% higher in PCB concentration than females, whereas females were 2% higher in Hg concentration than males. Males eliminating Hg from their bodies at a faster rate than females was a likely explanation for this discrepancy between the two contaminant types. Overall, the Hg and PCB concentrations in the summer flounder were relatively low, and therefore our findings also had implications for continued operation of the summer flounder fishery.

  11. Taxonomic recovery of the ant cricket Myrmecophilus albicinctus from M. americanus (Orthoptera, Myrmecophilidae).

    PubMed

    Komatsu, Takashi; Maruyama, Munetoshi

    2016-01-01

    Myrmecophilus americanus and Myrmecophilus albicinctus are typical myrmecophilous insects living inside ant nests. These species are ecologically important due to the obligate association with tramp ant species, including harmful invasive ant species. However, the taxonomy of these "white-banded ant crickets" is quite confused owing to a scarcity of useful external morphological characteristics. Recently, Myrmecophilus albicinctus was synonymized with Myrmecophilus americanus regardless of the apparent host use difference. To clarify taxonomical relationship between Myrmecophilus albicinctus and Myrmecophilus albicinctus, we reexamined morphological characteristics of both species mainly in the viewpoint of anatomy. Observation of genitalia parts, together with a few external body parts, revealed that Myrmecophilus albicinctus showed different tendency from them of Myrmecophilus americanus. Therefore, we recover Myrmecophilus albicinctus as a distinct species on the basis of the morphology. PMID:27408536

  12. Taxonomic recovery of the ant cricket Myrmecophilus albicinctus from M. americanus (Orthoptera, Myrmecophilidae)

    PubMed Central

    Komatsu, Takashi; Maruyama, Munetoshi

    2016-01-01

    Abstract Myrmecophilus americanus and Myrmecophilus albicinctus are typical myrmecophilous insects living inside ant nests. These species are ecologically important due to the obligate association with tramp ant species, including harmful invasive ant species. However, the taxonomy of these “white-banded ant crickets” is quite confused owing to a scarcity of useful external morphological characteristics. Recently, Myrmecophilus albicinctus was synonymized with Myrmecophilus americanus regardless of the apparent host use difference. To clarify taxonomical relationship between Myrmecophilus albicinctus and Myrmecophilus albicinctus, we reexamined morphological characteristics of both species mainly in the viewpoint of anatomy. Observation of genitalia parts, together with a few external body parts, revealed that Myrmecophilus albicinctus showed different tendency from them of Myrmecophilus americanus. Therefore, we recover Myrmecophilus albicinctus as a distinct species on the basis of the morphology. PMID:27408536

  13. Vertebral deformities in hatchery-reared and wild-caught juvenile Japanese flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Lü, Hongjian; Zhang, Xiumei; Fu, Mei; Xi, Dan; Su, Shengqi; Yao, Weizhi

    2015-01-01

    The present study compared vertebral deformities of hatchery-reared and wild-caught juvenile Japanese flounder, Paralichthys olivaceus. A total of 362 hatchery-reared flounder (total length 122.5-155.8 mm) were collected from three commercial hatcheries located in Yantai, East China, and 89 wild fish (total length 124.7-161.3 mm) were caught off Yangma Island near Yantai City (37°27'N, 121°36'E). All the fish were dissected, photographed, and images of the axial skeleton were examined for vertebral deformities. Compared with wild-caught flounder in which no deformed vertebrae were detected, 48 (13.3%) hatcheryreared fish had deformed vertebrae. The deformities were classified as compression, compression-ankylosis, and dislocation-ankylosis. The vertebral deformities were mainly localized between post-cranial vertebra 1 and 3, with vertebrae number 1 as the most commonly deformed. The causative factors leading to vertebral deformities in reared Japanese flounder may be related to unfavorable temperature conditions, inflammation, damage, or rupture to the intervertebral ligaments under rearing conditions. Furthermore, no significant difference in the total number of vertebral bodies was observed between wild-caught (38.8±0.4) and hatchery-reared flounder (38.1±0.9) ( P>0.05). However, the number of vertebral bodies of hatchery-reared and wild-caught flounder ranged from 35 to 39 and from 38 to 39, respectively.

  14. Identification of differentially expressed proteins of brain tissue in response to methamidophos in flounder (Paralichthys olivaceus).

    PubMed

    Peng, Hui-Fang; Bao, Xiao-Dong; Zhang, Yong; Huang, Lin; Huang, He-Qing

    2015-06-01

    Methamidophos (MAP), an organophosphorus pesticide used around the world, has been associated with a wide spectrum of toxic effects on organisms in the environment. In this study, the flounder Paralichthys olivaceus was subjected to 10 mg/L MAP for 72 h and 144 h, and the morphological and proteomic changes in the brain were observed, analyzed and compared with those in the non-exposed control group. Under the light microscope and transmission electron microscope, MAP had evidently induced changes in or damage to the flounder tissues. Gas chromatography analysis demonstrated that the MAP residues were significantly accumulated in the flounder brain tissues. Proteomic changes in the brain tissue were revealed using two-dimensional gel electrophoresis and 27 protein spots were observed to be significantly changed by MAP exposure. The results indicated that the regulated proteins were involved in immune and stress responses, protein biosynthesis and modification, signal transduction, organismal development, and 50% of them are protease. qRT-PCR was used to further detect the corresponding change of transcription. These data may be beneficial to understand the molecular mechanism of MAP toxicity in flounder, be very useful for MAP-resistance screening in flounder culture. According to our results and analyzing, heat shock protein 90 (HSP90) and granzyme K (GzmK) had taken important part in immune response to MAP-stress and could be biomarkers for MAP-stress in flounder.

  15. Expression Analysis of Visual Arrestin gene during Ocular Development of Olive Flounder (Paralichthys olivaceus)

    PubMed Central

    Yang, Hyun; Lee, Young Mee; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Hwang, In Joon; Kim, Sung Yeon; Lee, Jeong-Ho

    2013-01-01

    Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin. PMID:25949138

  16. Characterization and functional classification of American lobster (Homarus americanus) immune factor transcripts.

    PubMed

    Clark, K Fraser

    2014-11-01

    The American lobster (Homarus americanus) is the most important commercially exploited marine species in Canada. Very little is known about the H. americanus molecular humoral immune response or how to determine if a seemingly healthy lobster is infected with a pathogen. The goal of this work is to characterize several important H. americanus immune genes as well as highlight and classify hundreds of others into functional immune groups. The protein sequence of H. americanus acute phase serum amyloid protein A (SAA) was found to be similar to that of vertebrate SAA, and is likely a good clinical marker for immune activation in lobsters and some crustaceans. Additionally, only one gene, Trypsin 1b, was found to be differentially regulated during bacterial, microparasitic and viral challenges in lobster and is likely critical for the activation of the H. americanus immune response. Bioinformatic analysis was used to functionally annotate, 263 H. americanus immune genes and identify the few shared patterns of differential gene expression in lobsters in response to bacterial, parasitic and viral challenge. Many of the described immune genes are biomarker candidates which could be used as clinical indicators for lobster health and disease. Biomarkers can facilitate early detection of pathogens, or anthropomorphic stressors, so that mitigation strategies can be developed in order to prevent the devastating economic losses that have occurred in Southern New England, USA. This work is contributes to further our understanding of how the lobster immune system works and how it can be used to maintain the health and sustainability of the overall American lobster fishery.

  17. Neuropeptidergic Signaling in the American Lobster Homarus americanus: New Insights from High-Throughput Nucleotide Sequencing.

    PubMed

    Christie, Andrew E; Chi, Megan; Lameyer, Tess J; Pascual, Micah G; Shea, Devlin N; Stanhope, Meredith E; Schulz, David J; Dickinson, Patsy S

    2015-01-01

    Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysis-triggering hormone, red pigment concentrating hormone

  18. Characterization and functional classification of American lobster (Homarus americanus) immune factor transcripts.

    PubMed

    Clark, K Fraser

    2014-11-01

    The American lobster (Homarus americanus) is the most important commercially exploited marine species in Canada. Very little is known about the H. americanus molecular humoral immune response or how to determine if a seemingly healthy lobster is infected with a pathogen. The goal of this work is to characterize several important H. americanus immune genes as well as highlight and classify hundreds of others into functional immune groups. The protein sequence of H. americanus acute phase serum amyloid protein A (SAA) was found to be similar to that of vertebrate SAA, and is likely a good clinical marker for immune activation in lobsters and some crustaceans. Additionally, only one gene, Trypsin 1b, was found to be differentially regulated during bacterial, microparasitic and viral challenges in lobster and is likely critical for the activation of the H. americanus immune response. Bioinformatic analysis was used to functionally annotate, 263 H. americanus immune genes and identify the few shared patterns of differential gene expression in lobsters in response to bacterial, parasitic and viral challenge. Many of the described immune genes are biomarker candidates which could be used as clinical indicators for lobster health and disease. Biomarkers can facilitate early detection of pathogens, or anthropomorphic stressors, so that mitigation strategies can be developed in order to prevent the devastating economic losses that have occurred in Southern New England, USA. This work is contributes to further our understanding of how the lobster immune system works and how it can be used to maintain the health and sustainability of the overall American lobster fishery. PMID:24981290

  19. Neuropeptidergic Signaling in the American Lobster Homarus americanus: New Insights from High-Throughput Nucleotide Sequencing.

    PubMed

    Christie, Andrew E; Chi, Megan; Lameyer, Tess J; Pascual, Micah G; Shea, Devlin N; Stanhope, Meredith E; Schulz, David J; Dickinson, Patsy S

    2015-01-01

    Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysis-triggering hormone, red pigment concentrating hormone

  20. Neuropeptidergic Signaling in the American Lobster Homarus americanus: New Insights from High-Throughput Nucleotide Sequencing

    PubMed Central

    Christie, Andrew E.; Chi, Megan; Lameyer, Tess J.; Pascual, Micah G.; Shea, Devlin N.; Stanhope, Meredith E.; Schulz, David J.; Dickinson, Patsy S.

    2015-01-01

    Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysis-triggering hormone, red pigment concentrating hormone

  1. Gonadal Transcriptome Analysis in Sterile Double Haploid Japanese Flounder

    PubMed Central

    Wang, Guixing; Jiang, Hongbo; Wang, Yufen; Sun, Zhaohui; Jiang, Xiufeng; Yu, Qinghai; Liu, Haijin

    2015-01-01

    Sterility is a serious problem that can affect all bionts. In teleosts, double haploids (DHs) induced by mitogynogenesis are often sterile. This sterility severely restricts the further application of DHs for production of clones, genetic analysis, and breeding. However, sterile DH individuals are good source materials for investigation of the molecular mechanisms of gonad development, especially for studies into the role of genes that are indispensable for fish reproduction. Here, we used the Illumina sequencing platform to analyze the transcriptome of sterile female DH Japanese flounder in order to identify major genes that cause sterility and to provide a molecular basis for an intensive study of gonadal development in teleosts. Through sequencing, assembly, and annotation, we obtained 52,474 contigs and found that 60.7% of these shared homologies with existing sequences. A total of 1225 differentially expressed unigenes were found, including 492 upregulated and 733 downregulated genes. Gene Ontology and KEGG analyses showed that genes showing significant upregulation, such as CYP11A1, CYP11B2, CYP17, CYP21, HSD3β, bcl2l1, and PRLR, principally correlated with sterol metabolic process, steroid biosynthetic process, and the Jak-stat signaling pathway. The significantly downregulated genes were primarily associated with immune response, antigen processing and presentation, cytokine–cytokine receptor interaction, and protein digestion and absorption. Using a co-expression network analysis, we conducted a comprehensive comparison of gene expression in the gonads of fertile and sterile female DH Japanese flounder. Identification of genes showing significantly different expression will provide further insights into DH reproductive dysfunction and oocyte maturation processes in teleosts. PMID:26580217

  2. Gonadal Transcriptome Analysis in Sterile Double Haploid Japanese Flounder.

    PubMed

    Zhang, Xiaoyan; Hou, Jilun; Wang, Guixing; Jiang, Hongbo; Wang, Yufen; Sun, Zhaohui; Jiang, Xiufeng; Yu, Qinghai; Liu, Haijin

    2015-01-01

    Sterility is a serious problem that can affect all bionts. In teleosts, double haploids (DHs) induced by mitogynogenesis are often sterile. This sterility severely restricts the further application of DHs for production of clones, genetic analysis, and breeding. However, sterile DH individuals are good source materials for investigation of the molecular mechanisms of gonad development, especially for studies into the role of genes that are indispensable for fish reproduction. Here, we used the Illumina sequencing platform to analyze the transcriptome of sterile female DH Japanese flounder in order to identify major genes that cause sterility and to provide a molecular basis for an intensive study of gonadal development in teleosts. Through sequencing, assembly, and annotation, we obtained 52,474 contigs and found that 60.7% of these shared homologies with existing sequences. A total of 1225 differentially expressed unigenes were found, including 492 upregulated and 733 downregulated genes. Gene Ontology and KEGG analyses showed that genes showing significant upregulation, such as CYP11A1, CYP11B2, CYP17, CYP21, HSD3β, bcl2l1, and PRLR, principally correlated with sterol metabolic process, steroid biosynthetic process, and the Jak-stat signaling pathway. The significantly downregulated genes were primarily associated with immune response, antigen processing and presentation, cytokine-cytokine receptor interaction, and protein digestion and absorption. Using a co-expression network analysis, we conducted a comprehensive comparison of gene expression in the gonads of fertile and sterile female DH Japanese flounder. Identification of genes showing significantly different expression will provide further insights into DH reproductive dysfunction and oocyte maturation processes in teleosts. PMID:26580217

  3. Polychlorinated biphenyl (PCB) effects, PCB congener distributions, and cytochrome P-450 regulation in fish

    SciTech Connect

    Elskus, A.A.

    1992-01-01

    PCB studies were conducted in gonadally mature winter flounder (Pseudopleuronectes americanus) collected from spawning grounds with different degrees of PCB contamination. Flounder from Fox Is., RI, Gaspee Pt., RI and New Bedford, MA (US) had mean hepatic PCB concentrations between 1.6-10.6, 3.8-17.7, and 58-333 {mu}g PCB/g dry liver, respectively. PCB congener distributions in these flounder indicated that PCB disposition was not influenced by reproductive condition, that these migratory fish accumulate a significant portion of their PCB body burden during residence in their spawning grounds, and that flounder selectively metabolize congeners with adjacent meta-, para-unsubstituted carbon atoms. The most significant reproductive effect of PCBs in flounder was a 32 to 52% decrease in the mean ovarian size of highly contaminated females from New Bedford. Plasma levels of the sex hormones, estradiol and testosterone, and of the egg yolk precursor, vitellogenin, as well as hepatic estradiol metabolism (measured as estradiol 2-hydroxylase), showed no relationship to hepatic PCB concentration. PCB effects on flounder P4501A indicated that hepatic PCB concentrations as low as 0.9 {mu}g PCB/g were associated with decreased P4501A catalytic efficiency. Additional suppression of flounder P4501A by estrogens was suggested by depressed levels of P4501A messenger RNA, protein and catalytic activity in highly contaminated female fish with high estradiol titers. The mechanism of P4501A suppression by estrogens was studied in killifish (Fundulus heteroclitus) treated wit {beta}-napthoflavone ({beta}-NF), a P4501A inducer, and/or estradiol.

  4. Cortisol is involved in temperature-dependent sex determination in the Japanese flounder.

    PubMed

    Yamaguchi, Toshiya; Yoshinaga, Norifumi; Yazawa, Takashi; Gen, Koichiro; Kitano, Takeshi

    2010-08-01

    In vertebrates, sex is normally determined by genotype. However, in poikilothermal vertebrates, including reptiles, amphibians, and fishes, sex determination is greatly influenced by environmental factors, such as temperature. Little is known about the molecular mechanisms underlying environmental sex determination in these species. The Japanese flounder (Paralichthys olivaceus) is a teleost fish with an XX/XY sex determination system. However, XX flounder can be induced to develop into predominantly either phenotypic females or males, by rearing at 18 or 27 C, respectively, during the sex differentiation period. Therefore, the flounder provides an excellent model to study the molecular mechanisms underlying temperature-dependent sex determination. We previously showed that an aromatase inhibitor, an antiestrogen, and 27 C treatments cause masculinization of XX flounder, as well as suppression of mRNA expression of ovary-type aromatase (cyp19a1), a steroidogenic enzyme responsible for the conversion of androgens to estrogens in the gonads. Furthermore, estrogen administration completely inhibits masculinization by these treatments, suggesting suppression of cyp19a1 mRNA expression, and the resultant estrogen biosynthesis may trigger masculinization of the XX flounder induced by high water temperature. Here, we demonstrated that cortisol causes female-to-male sex reversal by directly suppressing cyp19a1 mRNA expression via interference with cAMP-mediated activation and that metyrapone (an inhibitor of cortisol synthesis) inhibits 27 C-induced masculinization of XX flounder. Moreover, cortisol concentrations in 27 C-reared juveniles were significantly higher than in 18 C-reared fishes during sexual differentiation. These results strongly suggest that masculinization by high water temperature is ascribable to elevation of cortisol concentration during gonadal sex differentiation in the flounder.

  5. SARCOCYSTIS URSUSI, N. SP (APICOMPLEXA; SARCOCYSTIDAE) FROM THE BLACK BEAR (URSUS AMERICANUS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infection with Sarcocystis species is common in hervibores but is rare in bears. Histological sections of 374 black bears (Ursus americanus) from Pennsylvania were examined for sarcocysts. A total of 3 sarcocysts were found in 3 bears, 1 sarcocyst per section. Sarcocysts from 2 bears were considered...

  6. Potential Vertical Transmission of Winter Ticks (Dermacentor albipictus) from Moose (Alces americanus) Dams to Neonates.

    PubMed

    Severud, William J; DelGiudice, Glenn D

    2016-01-01

    North American moose (Alces americanus) frequently become infested with winter ticks (Dermacentor albipictus). During capture of neonatal moose in northeastern Minnesota, US, in May-June 2013 and 2014, we recovered adult ticks from neonates, presumably vertically transferred from dams, heretofore, not documented. Infestations on neonates may have population-level implications.

  7. 77 FR 76950 - Fisheries of the Northeastern United States; Summer Flounder, Scup, and Black Sea Bass Fisheries...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-31

    ... limit. Following publication, an error was identified in the commercial summer flounder quota and recreational harvest limit. This rule corrects that error. DATES: Effective December 26, 2012, through December... an error in the commercial summer flounder quota and recreational harvest limit, which...

  8. Immigration, settlement and mortality of flounder ( Paralichthys olivaceus) larvae and juveniles in a nursery ground, Shijiki bay, Japan

    NASA Astrophysics Data System (ADS)

    Tanaka, M.; Goto, T.; Tomiyama, M.; Sudo, H.

    The occurrence, distribution and abundance of larvae and juveniles of a Japanese flounder, Paralichthys olivaceus, were investigated in Shijiki Bay, south-western Japan, from 1982 through 1987, with special reference to immigration, settlement and mortality in this nursery ground. Sampling for pelagic larvae and settling and settled juveniles revealed that immigration and settlement occur during the late phase of metamorphosis, when flounders immigrate from coastal waters and settle in the near-shore sandy nursery area. Immigration of metamorphosing larvae begins in early April and continues until early June. The recruitment of flounders into the nursery ground was found to occur in several distinct phases, coinciding with spring tides. The semi-lunar periodicity found in the immigration of flounders may be due to the combination of landward tidal currents during spring tides and the tide-related vertical movements of metamorphosing flounders. Population size appeared to decrease rapidly during the first week after settlement, when lengths ranged from 11 to 14 mm. Field evidence demonstrated that heavy mortality may occur during the early phases of settlement, the extent depending on annual flounder densities and food abundance. Cannibalism of early-settled larger flounders on late-settled smaller flounders seemed to occur frequently in the nursery ground, and was more likely to occur under conditions of food deficiency and higher population densities. Thus, predation related t starvation could be one of the most important future research targets in determining recruitment dynamics in nursery areas.

  9. 77 FR 24151 - Fisheries of the Northeastern United States; Summer Flounder, Scup, and Black Sea Bass Fisheries...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-23

    ... INFORMATION: On December 30, 2011 (76 FR 82189), NMFS published interim specifications for the summer flounder... revised quota setting procedures for the FMP (67 FR 6877, February 14, 2002), summer flounder overages are... following Federal Register documents: 77 FR 14481 (March 12, 2012); and 77 FR 19951 (April 3, 2012). Table...

  10. Fish health and environmental health.

    PubMed Central

    Murchelano, R A

    1990-01-01

    Surveys conducted to evaluate the health of marine-bottom fishes have been conducted in the eastern and western North Atlantic for the past 15 years, usually in conjunction with fish stock assessment cruises. The health of the fish sampled was evaluated using certain integumental and skeletal lesions and anomalies as markers to signify compromised health status. The results of these surveys indicate that fish health is poorer in coastal waters that have been anthropogenically degraded. Monitoring programs to determine the status and trends in levels of inorganic and organic contaminants in fish tissue and sediments have disclosed high levels of chemical contaminants in several coastal areas of the northeastern United States. Histopathological examinations of liver tissues of winter flounder, Pseudopleuronectes americanus, from Boston Harbor, one of the more chemically contaminated sites, has revealed a high prevalence of hepatocarcinoma. PMID:2401261

  11. Differential Expression Patterns of Crystallin Genes during Ocular Development of Olive Flounder (Paralichthys olivaceus)

    PubMed Central

    Yang, Hyun; Lee, Young Mee; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul Ji; Park, Jong Won; Hwang, In Joon; Kim, Sung Yeon; Lee, Jeong Ho

    2012-01-01

    Olive flounder Paralichthys olivaceus is one of the most widely cultured fish species in Korea. Although olive flounder receive attention from aquaculture and fisheries and extensive research has been conducted eye morphological change in metamorphosis, but little information was known to molecular mechanism and gene expression of eye development- related genes during the early part of eye formation period. For the reason of eyesight is the most important sense in flounder larvae to search prey, the screening and identification of expressed genes in the eye will provide useful insight into the molecular regulation mechanism of eye development in olive flounder. Through the search of an olive flounder DNA database of expressed sequence tags (EST), we found a partial sequence that was similar to crystallin beta A1 and gamma S. Microscopic observation of retinal formation correspond with the time of expression of the crystallin beta A1 and gamma S gene in the developmental stage, these result suggesting that beta A1 and gamma S play a vital role in the remodeling of the retina during eye development. The expression of crystallin beta A1 and gamma S were obviously strong in eye at all tested developing stage, it is also hypothesized that crystallin acts as a molecular chaperone to prevent protein aggregation during maturation and aging in the eye. PMID:25949104

  12. N-Terminal region is responsible for chemotaxis-inducing activity of flounder IL-8.

    PubMed

    Kurata, Osamu; Wada, Shinpei; Matsuyama, Tomomasa; Sakai, Takamitsu; Takano, Tomokazu

    2014-06-01

    The objective of this study was to locate the functional region responsible for the chemotaxis-inducing activity of flounder interleukin 8 (IL-8), which lacks the glutamic acid-leucine-arginine (ELR) motif essential for the induction of neutrophil migration by mammalian IL-8. Using a human cell line, we produced a secretory recombinant protein of flounder IL-8, and analyzed its chemotaxis-inducing activity on leukocytes collected from the flounder kidney. The recombinant IL-8 induced significant migration in neutrophils, which were morphologically and functionally characterized. Using the Edman degradation method, the N-terminal amino acid sequence of rIL-8 was identified as VSLRSLGV. To examine the significance of the N-terminal region for the bioactivity of flounder IL-8, we prepared several recombinant proteins that containing mutations at the N-terminus. Modification of three residues (residues 9-11: serine-leucine-histidine) corresponding in position to the ELR motif in mammalian IL-8 did not reduce its chemotaxis-inducing activity. However, deletion of the first six or more residues significantly reduced its chemotaxis-inducing activity. We propose that residue 6 (leucine) at the N-terminus is important for the chemotaxis-inducing activity of flounder IL-8.

  13. Population structure of flounder (Platichthys flesus) in the Baltic Sea: differences among demersal and pelagic spawners.

    PubMed

    Florin, A-B; Höglund, J

    2008-07-01

    We found significant population structure and isolation by distance among samples of flounder (Platichthys flesus) in the Baltic, Kattegat and Skagerrak seas using microsatellite genetic markers. This pattern was almost entirely due to a difference between flounder that have demersal spawning in the northern Baltic, as compared to pelagic spawners in the southern Baltic and on the west coast of Sweden. Among demersal spawners we found neither genetic differentiation nor any isolation by distance among sampling sites. We speculate that demersal flounder are descendants of a population that colonized the Baltic previous to pelagic spawners. The demersal flounder may thus have had longer time to adapt to the low salinity in the Baltic, and accordingly display egg characteristics that make it possible to reproduce at the low salinity levels in the northern Baltic. Among pelagic spawners significant isolation by distance was detected. Pelagic spawners have previously been shown to display clinal variation in egg size, which allows them to float also at the moderate salinity levels up to the region north of the island Bornholm. Management units for harvesting should ideally be based on true biological populations, and for the commercially important flounder up to 15 different management stocks in the Baltic have been suggested. We could not find a population genetic foundation for such a high number of management units, and our data suggest three management units: the northern Baltic (demersal populations), southern Baltic with the Oresund straits and the most northwestern sampling sites (Skagerrak, Kattegat and North Sea).

  14. Production and verification of a 2nd generation clonal group of Japanese flounder, Paralichthys olivaceus

    PubMed Central

    Hou, Jilun; Zhang, Xiaoyan; Wang, Yufen; Sun, Zhaohui; Si, Fei; Jiang, Xiufeng; Liu, Haijin

    2016-01-01

    Clonal fishes are useful tools in biology and aquaculture studies due to their isogenicity. In Japanese flounder (Paralichthys olivaceus), a group of homozygous clones was created by inducing meiogynogenesis in eggs from a mitogynogenetic homozygous diploid. As the clones reached sexual maturity, meiogynogenesis was again induced in order to produce a 2nd generation clonal group of Japanese flounder. After 3 months, there were 611 healthy, surviving individuals. Twenty-four microsatellite markers, that covered all the linkage groups of Japanese flounder, were used to identify the homozygosity of the 2nd generation clones; no heterozygous locus was detected. This indicates that the production of a 2nd generation clonal group of Japanese flounder was successful. Restriction-site DNA associated sequencing at the genomic level also confirmed the homozygosity and clonality of the 2nd generation clonal group. Furthermore, these 2nd generation clones had a small coefficient of variation for body shape indices at 210 days of age and showed a high degree of similarity in body characteristics among individuals. The successful production of 2nd generation clones has laid the foundation for the large-scale production of clonal Japanese flounder. PMID:27767055

  15. Characterization, tissue distribution, and expression of neuropeptide Y in olive flounder Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Tan, Xungang; Du, Shaojun; Sun, Wei; You, Feng; Zhang, Peijun

    2015-05-01

    Neuropeptide Y (NPY) is a 36-amino acid peptide of the neuropeptide Y family that plays key roles in the regulation of food intake. In this study, we focused on NPY mRNA expression changes around feeding time and during food deprivation in olive flounder. The olive flounder NPY mRNA levels were analyzed in different tissues and a high level of expression was detected in the brain. We also demonstrated a correlation between NPY expression levels in the brain and feeding schedule. NPY expression levels in olive flounder maintained on a daily scheduled feeding regimen increased shortly before feeding and decreased after the scheduled feeding time. Compared with the -1 h group before feeding, NPY expression in the 3 h group after feeding decreased significantly ( P<0.05). Food deprivation led to an 81.7% decrease in NPY mRNA levels in the 24 h fasted group ( P<0.05) and a 91.7% decrease in the 48 h fasted group ( P<0.05). Therefore, our study demonstrates that NPY expression is associated with food intake in olive flounder. This result reveals the function of NPY in regulating food intake and its potential importance in olive flounder aquaculture.

  16. Food intake of young cod and flounder in a shallow bay on the Swedish west coast

    NASA Astrophysics Data System (ADS)

    Pihl, L.

    The importance of shallow soft bottom areas as nursery grounds for I-ground cod, Gadus morhua, and I- to IV-group flounder, Platichthys flesus, was investigated in a 20 000 m 2 bay, 0 to 1 m deep, on the Swedish west coast. To estimate the amount of visiting fish and their food intake the bay was enclosed with a net once a month from July to November 1979 and from April to November 1980. At each occasion about 60 cods and 8 to 50 flounders were caught. The total biomass of the cod and flounder was 1 to 14 kg and 2 to 13 kg wet weight, respectively. The cod preyed on mobile epifauna such as Carcinus maenas, Crangon crangon and Pomatoschistus spp., whereas the flounder was eating infauna mainly Mya arenaria, Cardium spp. and Nereis spp. The food intake within the area was calculated to approximate 35 and 10 kg wet weight from April to November for cod and flounder, respectively.

  17. Analysis of IL-2-like factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

    NASA Astrophysics Data System (ADS)

    Wu, Riqin; Zhang, Peijun; Li, Jun; Xu, Yongli

    2005-03-01

    To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder, Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes ( P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2, (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%.

  18. Comparisons of liver proteomes in the European flounder Platichthys flesus from three contrasted estuaries

    NASA Astrophysics Data System (ADS)

    Galland, Claire; Dupuy, Célie; Capitaine, Carole; Auffret, Michel; Quiniou, Louis; Laroche, Jean; Pichereau, Vianney

    2013-01-01

    Estuaries are important areas highly susceptible to anthropogenic degradations like pollution. Estuarine species have thus to cope with many types of constraints depending on the estuaries' characteristics. The European flounder Platichthys flesus is considered as a sentinel species for the monitoring of estuarine water quality. In this study, juvenile flounders (0+ group) were sampled from three contrasted Channel estuaries, i.e. the Seine, the Canche and the Tamar, and we characterized their liver proteomes by using a two-dimensional electrophoresis based proteomic approach. We showed that 27 protein spots differentially accumulated between the 3 populations. Six of these proteins were identified by MALDI TOF-TOF mass spectrometry. Flounders from the Seine and from the Tamar, two highly polluted estuaries, displayed common differences, i.e. an increase of the energetic- and the glutathione-metabolism. The most accumulated protein in the Seine's samples (6.7-fold) was a Vitelline Membrane Outer layer protein 1 homolog, suggesting oogenesis deregulation in these juvenile (sexually immature) flounders. Future works applying this kind of proteomic approach on flounders experimentally exposed to conditions that mimic environmental constraints will help to better understand the significance of these environmental proteomic signatures.

  19. The toxic mechanism of high lethality of herbicide butachlor in marine flatfish flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Guo, Huarong; Yin, Licheng; Zhang, Shicui; Feng, Wenrong

    2010-09-01

    The toxic mechanism of herbicide butachlor to induce extremely high lethality in marine flatfish flounder, Paralichthys Olivaceus, was analyzed by histopathological examination, antioxidant enzymes activities and ATP content assay. Histopathological examination of gill, liver and kidney of exposed fishes showed that gill was a target organ of butachlor. The butachlor seriously impaired the respiration of gills by a series of lesions such as edema, lifting and detachment of lamellar epithelium, breakdown of pillar cells, and blood congestion. The dysfunction of gill respiration caused suffocation to the exposed flounder with extremely high acute lethality. Antioxidant enzyme activity assay of the in vitro cultured flounder gill (FG) cells exposed to butachlor indicated that butachlor markedly inhibited the antioxidant enzyme activities of Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX). Furthermore, along with the decline of antioxidant enzyme activities, ATP content in the exposed FG cells decreased, too. This infers that the oxidative stress induced by butachlor can inhibit the production of cellular ATP. Similar decrease of ATP content was also observed in the exposed flounder gill tissues. Taken together, as in FG cells, butachlor possibly induced a short supply of ATP in pillar cells by inhibiting the antioxidant enzyme activities and then affecting the contractibility of the pillar cells, which in turn resulted in the blood congestion and suffocation of exposed flounder.

  20. Biomarkers of exposure and effect in the Brazilian flounder Paralichthys orbignyanus (Teleostei: Paralichthyidae) from the Patos Lagoon estuary (Southern Brazil).

    PubMed

    Amado, Lílian Lund; Robaldo, Ricardo B; Geracitano, Laura; Monserrat, José M; Bianchini, Adalto

    2006-02-01

    Biomarkers of exposure (liver metallothionein-like proteins content and catalase and glutathione S-transferase activities) and effect (liver lipoperoxidation and blood cell DNA damage) of contaminants were analyzed in the Brazilian flounder Paralichthys orbignyanus from the Patos Lagoon estuary (Southern Brazil). Flounders were collected for a year in two sites: "Coroa do Boi" (polluted site) and "Saco do Justino" (non-polluted site). Results indicated that micronucleated cells frequency was the best biomarker to distinguish flounders from the two sites. Taken together, data from DNA damage analyses (micronucleus test and comet assay) indicated that flounders from the non-polluted site efficiently repaired the DNA breaks, contrary to those from the polluted site, which probably had their DNA repair system inhibited or exhausted. Furthermore, data from enzyme activities (catalase and GST) and lipid peroxidation indicated that flounders from the polluted site were under oxidative stress in summer and autumn.

  1. Ice-binding mechanism of winter flounder antifreeze proteins.

    PubMed Central

    Cheng, A; Merz, K M

    1997-01-01

    We have studied the winter flounder antifreeze protein (AFP) and two of its mutants using molecular dynamics simulation techniques. The simulations were performed under four conditions: in the gas phase, solvated by water, adsorbed on the ice (2021) crystal plane in the gas phase and in aqueous solution. This study provided details of the ice-binding pattern of the winter flounder AFP. Simulation results indicated that the Asp, Asn, and Thr residues in the AFP are important in ice binding and that Asn and Thr as a group bind cooperatively to the ice surface. These ice-binding residues can be collected into four distinct ice-binding regions: Asp-1/Thr-2/Asp-5, Thr-13/Asn-16, Thr-24/Asn-27, and Thr-35/Arg-37. These four regions are 11 residues apart and the repeat distance between them matches the ice lattice constant along the (1102) direction. This match is crucial to ensure that all four groups can interact with the ice surface simultaneously, thereby, enhancing ice binding. These Asx (x = p or n)/Thr regions each form 5-6 hydrogen bonds with the ice surface: Asn forms about three hydrogen bonds with ice molecules located in the step region while Thr forms one to two hydrogen bonds with the ice molecules in the ridge of the (2021) crystal plane. Both the distance between Thr and Asn and the ordering of the two residues are crucial for effective ice binding. The proper sequence is necessary to generate a binding surface that is compatible with the ice surface topology, thus providing a perfect "host/guest" interaction that simultaneously satisfies both hydrogen bonding and van der Waals interactions. The results also show the relation among binding energy, the number of hydrogen bonds, and the activity. The activity is correlated to the binding energy, and in the case of the mutants we have studied the number of hydrogen bonds. The greater the number of the hydrogen bonds the greater the antifreeze activity. The roles van der Waals interactions and the hydrophobic

  2. Expression of Vimentin Intermediate Filament for Vascular Development in Olive Flounder (Paralichthys olivaceus)

    PubMed Central

    Yang, Hyun; Lee, Jang-Wook; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Hwang, In Joon; Kim, Sung Yeon; Lee, Jeong-Ho

    2014-01-01

    Cardiovascular system is the primary organ to develop and reach a functional state, which underscores the essential role of the vasculature in the developing embryo. The vasculature is a highly specialized organ that functions in a number of key physiological works including the carrying of oxygen and nutrients to tissues. It is closely involved in the formation of heart, and hence it is essential for survival during the hatching period. The expression of genes involved during vascular development in the olive flounder (Paralichthys olivaceus) in the days after hatching is not fully understood. Therefore, we examined the expression patterns of genes activated during the development of flounder. Microscopic observations showed that formation of blood vessels is related to the expression of the vimentin gene. Also, the temporal expression patterns of this vimentin-like gene in the developmental stages and in the normal tissues of olive flounder. The purpose of this study was to examine the expression patterns of vimentin in normal tissues of the olive flounder and during the development of the vascular system in newly hatched olive flounders and HIF-1 plays a vital role in the formation of blood vessels during development. Vimentin expression was strong at the beginning of the development of blood vessels, and was present throughout all developmental stages. Our findings have important implications with respect to the roles of vimentin and HIF-1 in the development and evolution of the first blood vessels in olive flounder. Further studies are required to elucidate the vimentin-mediated hypoxic response signal transduction and to decipher the functional role of vimentin in developmental stages. PMID:25949178

  3. High doses of corticosteroid suppress resistance to Ichthyophonus in starry flounder

    USGS Publications Warehouse

    Perry, J.A.; Kocan, R.M.; Winton, J.R.; Hershberger, P.K.

    2004-01-01

    Application of pharmacological doses of the corticosteroid dexamethasone phosphate to starry flounder Platichthys stellatus resulted in a predisposition to clinical ichthyophoniasis and a progression from latent Ichthyophonus infections to patent, histologically identifiable infections. Among Ichthyophonus-challenged starry flounder, the prevalences of clinical infections and histologically identifiable infections were significantly greater in two groups that received dexamethasone (100% and 31%, respectively) than in the respective control groups (8% and 0%). Proliferation of Ichthyophonus infections in corticosteroid-treated groups may have resulted from suppression of the cellular immune response that typically follows corticosteroid application; however, further studies are needed to determine whether these effects occur at lower, physiological concentrations of corticosteroids.

  4. miR-17 is involved in Japanese Flounder (Paralichthys olivaceus) development by targeting the Cdc42 mRNA.

    PubMed

    Zhang, Hongmei; Fu, Yuanshuai; Shi, Zhiyi; Su, Yanfang; Zhang, Junling

    2016-01-01

    The expression patterns of 197 miRNAs during Japanese flounder metamorphic development were recently analyzed. miR-17 was differentially expressed during the metamorphic period of the Japanese flounder; however, the role of miR-17 in Japanese flounder development has remained elusive to date. Bioinformatics analysis showed that Cdc42 was a putative target of miR-17. Cdc42 is a gene related to cell adhesion, migration, polarity, cytokinesis, growth, actin cytoskeleton, microtubule dynamics and transcription factor activity; thus, Cdc42 may contribute to metamorphic development. In our study, overexpression of miR-17 in FEC cells suppressed Cdc42 expression. The luciferase reporter assay confirmed that Cdc42 was the target of miR-17. The Cdc42 cDNA from the Japanese flounder was cloned and characterized for the first time. The expression of miR-17 was found to be negatively correlated with Cdc42 mRNA expression during temporal development and in the tissues of adult Japanese flounders. These results indicated that the decrease in miR-17 contributed to the up-regulation of Cdc42 during Japanese flounder metamorphosis. Cdc42 gene expression was down-regulated by thyroid hormone during Japanese flounder metamorphosis, whereas miR-17 was significantly up-regulated by thyroid hormone during these stages. These results indicated that miR-17 was a negative regulator of Cdc42. PMID:26546744

  5. Microlichus americanus acariasis in saffron finches (Sicalis flaveola) with dermatitis and feather loss.

    PubMed

    Rettenmund, Christy L; Ossiboff, Robert J; McAloose, Denise; Knee, Wayne; Wade, Susan E; Paré, Jean A

    2015-05-01

    Over a 5-year period, 13 saffron finches (Sicalis flaveola) housed in mixed aviaries at the Bronx Zoo (Bronx, New York) were examined with feather loss and dermatitis, primarily affecting the nape, neck, and dorsum. Feather loss, hyperkeratosis, epidermal hyperplasia, and mixed granulocytic and mononuclear inflammation were identified in biopsies from live birds and tissue sections from postmortem specimens. In 10 of 13 cases, sections of arthropod parasites were seen histologically within feather follicles and along the surface of affected skin. Based on morphological characteristics, mites recovered from samples of formalin-fixed skin in 4 birds were identified as Microlichus americanus, an epidermoptid mite infrequently reported from wild birds and hippoboscid flies. Gross and histological lesions strongly implicate M. americanus as the cause of dermatitis affecting practically all saffron finches in the collection.

  6. External morphology of the cycliophoran dwarf male: a comparative study of Symbion pandora and S. americanus

    NASA Astrophysics Data System (ADS)

    Neves, Ricardo Cardoso; da Cunha, Marina Ribeiro; Funch, Peter; Wanninger, Andreas; Kristensen, Reinhardt Møbjerg

    2010-09-01

    Cycliophora is a recently described phylum to which only two species have been assigned so far, Symbion pandora and S. americanus. The cycliophoran life cycle is complex and alternates between asexual and sexual stages. Although not recognized as an entirely independent free-swimming stage when the phylum was first described, the dwarf male has a remarkably complex bodyplan albeit its very small size (approx. 30-40 μm in length). Aiming to increase the knowledge on the gross morphology of the cycliophoran dwarf male, specimens from S. pandora and S. americanus were analyzed by scanning electron microscopy. In both species, anterior and ventral ciliated fields, as well as paired lateral sensorial organs, were identified, thus confirming previous observations. However, new details are described herein such as the penial pouch that encloses the penis. We compare our findings on both Symbion species with the data currently available on other metazoan dwarf males.

  7. A microchannel confocal examination of arsenic speciation and distribution in Bufo americanus

    NASA Astrophysics Data System (ADS)

    Nearing, Michelle M.; Koch, Iris; Gordon, Robert A.; Reimer, Kenneth J.

    2016-05-01

    We have used confocal methods to examine the distribution and speciation of arsenic within amphibian tissue (Bufo americanus) from a contaminated mine site. The use of new microchannel technology permits a confocal, and energy-independent, examination of a given voxel within the amphibian tissue without the need for sectioning. We observe differences in arsenic concentration and speciation depending on tissue type, with the Eberth- Katschenko layer in particular containing Mn, Fe, Cu and Zn in addition to Ca and pentavalent arsenic.

  8. Sequence data mining in search of hookworm (Necator americanus) microRNAs.

    PubMed

    Kulkarni, Abhijeet P; Mittal, Smriti P K

    2016-09-30

    The new world hookworm, Necator americanus is a soil-transmitted nematode responsible for Necatoriasis (a type of helminthiasis) in hosts such as humans, dogs, and cats. N. americanus genome and transcriptome has been sequenced and a draft assembly analysis has been published highlighting protein coding genes and possible drug target proteins. Hookworm microRNA identification, annotations and their public release is yet to be attempted. The same is evident from lack of hookworm miRNA information in related popular public nucleotide sequence repositories such as miRBase, GenBank, WormBase etc. Therefore, in the present study we addressed these issues using EST and assembled transcript sequence information of hookworm. Using computational approaches, we identified three miRNAs precursor sequences and their mature forms. We also identified their potential targets from hookworm ESTs and transcripts, and from human transcriptome. Overall, the results indicate presence of nematode specific miRNA homologs in N. americanus and shades light on their putative targets in worm itself and the human host. PMID:27259664

  9. Sequence data mining in search of hookworm (Necator americanus) microRNAs.

    PubMed

    Kulkarni, Abhijeet P; Mittal, Smriti P K

    2016-09-30

    The new world hookworm, Necator americanus is a soil-transmitted nematode responsible for Necatoriasis (a type of helminthiasis) in hosts such as humans, dogs, and cats. N. americanus genome and transcriptome has been sequenced and a draft assembly analysis has been published highlighting protein coding genes and possible drug target proteins. Hookworm microRNA identification, annotations and their public release is yet to be attempted. The same is evident from lack of hookworm miRNA information in related popular public nucleotide sequence repositories such as miRBase, GenBank, WormBase etc. Therefore, in the present study we addressed these issues using EST and assembled transcript sequence information of hookworm. Using computational approaches, we identified three miRNAs precursor sequences and their mature forms. We also identified their potential targets from hookworm ESTs and transcripts, and from human transcriptome. Overall, the results indicate presence of nematode specific miRNA homologs in N. americanus and shades light on their putative targets in worm itself and the human host.

  10. Effects of Cellulolytic Ruminal Bacteria and of Cell Extracts on Germination of Euonymus americanus L. Seeds.

    PubMed

    Howard, Gary T; Elliott, Larry P

    1988-01-01

    In past attempts, the experimental germination of the seeds of Euonymus americanus L. in vitro has had little success. However, treatment of seeds with ruminal fluid containing viable microflora has been successful in stimulating germination. In the presence of the cellulolytic ruminal bacterium, Clostridium cellobioparum ATCC 15832, seeds of E. americanus were stimulated to germinate. Subsequent studies were designed to determine whether the bacterium synthesized a cellulolytic enzyme responsible for initiating germination. The cell-free endocellulase from C. cellobioparum induced germination of the seeds. To support the hypothesis that the endocellulase from C. cellobioparum was responsible for triggering germination, a 1,4-beta-d-glucan glucanohydrolase (EC 3.2.1.4) from Penicillum funiculosum was used to treat the seeds. In addition, no germination was obtained from seeds treated with a commercial exocellulase enzyme. Also, Ruminococcus flavefaciens FD-1 was found to initiate germination of E. americanus seeds. Thus, cellulase activity is indicated in the degradation of the testa of the seed, allowing imbibition and germination.

  11. Screening of eye-position related genes with DD-RT-PCR and RDA in the hybrids between Japanese flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus

    NASA Astrophysics Data System (ADS)

    Chen, Yanjie; Zhang, Quanqi; Qi, Jie; Sun, Yeying; Zhong, Qiwang; Wang, Xubo; Wang, Zhigang; Li, Shuo; Li, Chunmei

    2009-02-01

    Flatfish or flounder moves one eye to change body proportion into vertebral asymmetry during metamorphosis, during which some become sinistral while others dextral. However, the mechanism behinds the eye-position has not been well understood. In this research, hybrids between Japanese flounder(♀) and stone flounder (♂) show mixed eye-location in both dextral type and sinistral type, and thus become good samples for studying the eye-migration. mRNAs from pro-metamorphosis sinistral and dextral hybrids larvae were screened with classical differential display RT-PCR (DD-RT-PCR) and representational difference analysis of cDNA (cDNA-RDA); 30 and 47 putative fragments were isolated, respectively. The cDNA fragments of creatine kinase and trypsinogen 2 precursor genes isolated by cDNA-RDA exhibited eye-position expression patterns during metamorphosis. However, none of the fragments was proved to be related to flatfishes’ eye-position specifically. Therefore, further studies and more sensitive gene isolated methods are needed to solve the problems.

  12. USING WINTER FLOUNDER GROWTH RATES AND STABLE ISOTOPES TO ASSESS HABITAT QUALITY

    EPA Science Inventory

    We used winter flounder growth rates and stable isotopes to assess habitat quality across an anthropogenic gradient in Narragansett Bay, Rhode Island. Cages (1 m2) were placed in the Providence River which had the highest nutrient concentrations and greatest development, Prudence...

  13. RELATIONSHIPS BETWEEN JUVENILE WINTER FLOUNDER AND MULTIPLE-SCALE HABITAT VARIATION IN NARRAGANSETT BAY, RHODE ISLAND

    EPA Science Inventory

    A rapid random-sampling method was used to relate densities of juvenile winter flounder to multiple scales of habitat variation in Narragansett Bay and two nearby coastal lagoons in Rhode Island. We used a 1-m beam trawl with attached video camera, continuous GPS track overlay, ...

  14. THE PATHOBIOLOGY OF 17B-ESTRADIOL IN SUMMER FLOUNDER, PARALICHTYS DENTATUS

    EPA Science Inventory

    Estradiol has been shown to cause increased vitellogenin (VtG) concentrations in male fish. The intent of this study was to evaluate the pathobiology associated with exposure to 17 -estradiol (E2) on liver, gonad, and kidney tissues of summer flounder, Paralichthys dentatus. Juve...

  15. 50 CFR Figure 6 to Part 223 - TED Extension in Summer Flounder Trawl

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 9 2011-10-01 2011-10-01 false TED Extension in Summer Flounder Trawl 6 Figure 6 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES...

  16. 50 CFR Figure 6 to Part 223 - TED Extension in Summer Flounder Trawl

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 10 2012-10-01 2012-10-01 false TED Extension in Summer Flounder Trawl 6 Figure 6 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE MARINE MAMMALS THREATENED MARINE AND ANADROMOUS SPECIES...

  17. 75 FR 52890 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-30

    ... for the 2010 calendar ] year was set equal to 13,278,001 lb (6,023 mt) (74 FR 67978, December 22, 2009... to advise Massachusetts that the quota has been harvested and to advise vessel permit holders and dealer permit holders that no commercial quota is available for landing summer flounder in...

  18. Differential regulation of taurine biosynthesis in rainbow trout and Japanese flounder.

    PubMed

    Wang, Xuan; He, Gen; Mai, Kangsen; Xu, Wei; Zhou, Huihui

    2016-02-16

    Animals have varied taurine biosynthesis capability, which was determined by activities of key enzymes including cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSD). However, whether CDO and CSD are differentially regulated across species remains unexplored. In the present study, we examined the regulations of CDO and CSD in rainbow trout and Japanese flounder, the two fish species with high and low taurine biosynthesis ability respectively. Our results showed that the expression of CDO was lower in rainbow trout but more responsive to cysteine stimulation compared to that in Japanese flounder. On the other hand, both the expression and catalytic efficiency (k(cat)) of CSD were higher in rainbow trout than those of Japanese flounder. A three-residue substrate recognition motif in rainbow trout CSD with sequence of F126/S146/Y148 was identified to be responsible for high k(cat), while that with sequence of F88/N108/F110 in Japanese flounder led to low k(cat), as suggested by site-directed mutagenesis studies. In summary, our results determined new aspects of taurine biosynthesis regulation across species.

  19. Stabel Carbon and Oxygen Isotope Ratios of Otoliths from Juvenile and Adult Winter Flounder

    EPA Science Inventory

    This study was designed to determine if stable carbon (13C) and oxygen (18O) isotope ratios in otoliths could be used to differentiate the locations that serve as important nursery areas for winter flounder along the Rhode Island, USA coastline. In recent years the populations ...

  20. Differential regulation of taurine biosynthesis in rainbow trout and Japanese flounder

    PubMed Central

    Wang, Xuan; He, Gen; Mai, Kangsen; Xu, Wei; Zhou, Huihui

    2016-01-01

    Animals have varied taurine biosynthesis capability, which was determined by activities of key enzymes including cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSD). However, whether CDO and CSD are differentially regulated across species remains unexplored. In the present study, we examined the regulations of CDO and CSD in rainbow trout and Japanese flounder, the two fish species with high and low taurine biosynthesis ability respectively. Our results showed that the expression of CDO was lower in rainbow trout but more responsive to cysteine stimulation compared to that in Japanese flounder. On the other hand, both the expression and catalytic efficiency (kcat) of CSD were higher in rainbow trout than those of Japanese flounder. A three-residue substrate recognition motif in rainbow trout CSD with sequence of F126/S146/Y148 was identified to be responsible for high kcat, while that with sequence of F88/N108/F110 in Japanese flounder led to low kcat, as suggested by site-directed mutagenesis studies. In summary, our results determined new aspects of taurine biosynthesis regulation across species. PMID:26880478

  1. 50 CFR 648.64 - Yellowtail flounder sub-ACLs and AMs for the scallop fishery.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Yellowtail flounder sub-ACLs and AMs for the scallop fishery. 648.64 Section 648.64 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures for...

  2. 50 CFR 648.64 - Yellowtail flounder sub-ACLs and AMs for the scallop fishery.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 10 2011-10-01 2011-10-01 false Yellowtail flounder sub-ACLs and AMs for the scallop fishery. 648.64 Section 648.64 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures for...

  3. 50 CFR 648.64 - Yellowtail flounder sub-ACLs and AMs for the scallop fishery.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Yellowtail flounder sub-ACLs and AMs for the scallop fishery. 648.64 Section 648.64 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures for...

  4. 50 CFR 648.107 - Conservation equivalent measures for the summer flounder fishery.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 8 2010-10-01 2010-10-01 false Conservation equivalent measures for the summer flounder fishery. 648.107 Section 648.107 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures...

  5. 50 CFR 648.107 - Conservation equivalent measures for the summer flounder fishery.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Conservation equivalent measures for the summer flounder fishery. 648.107 Section 648.107 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures...

  6. CONSUMPTIONS RATES OF SUMMER FLOUNDER LARVAE ON ROTIFER AND BRINE SHRIMP PREY DURING LARVAL REARING

    EPA Science Inventory

    Larval summer flounder Paralichthys dentatus were hatched and reared through metamorphosis in the laboratory. At several points in the rearing cycle, larvae were removed from their rearing chambers and placed in small bowls, where they were fed known quantities of the rotifer Bra...

  7. 50 CFR 648.64 - Yellowtail flounder sub-ACLs and AMs for the scallop fishery.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Yellowtail flounder sub-ACLs and AMs for the scallop fishery. 648.64 Section 648.64 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE...

  8. 78 FR 78786 - Fisheries of the Northeastern United States; 2014 Commercial Summer Flounder Quota Adjustments

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-27

    ... result of a recent stock assessment and a recommendation by the Mid-Atlantic Fishery Management Council..., 2012; 77 FR 76942). These specifications included the initial state allocations, as well as the state... overages from fishing year 2013, through October 31, 2013. As a result, the 2014 summer flounder...

  9. Differentially expressed genes after viral haemorrhagic septicaemia virus infection in olive flounder (Paralichthys olivaceus).

    PubMed

    Hwang, Jee Youn; Kwon, Mun-Gyeong; Seo, Jung Soo; Do, Jung Wan; Park, Myoung-Ae; Jung, Sung-Hee; Ahn, Sang Jung

    2016-09-25

    A strain of viral haemorrhagic septicaemia virus (VHSV) was isolated from cultured olive flounder (Paralichthys olivaceus) during epizootics in South Korean. This strain showed high mortality to olive flounder in in vivo challenge experiment. The complete genomic RNA sequences were determined and phylogenetic analysis of the amino acid sequences of glycoprotein revealed that this isolate was grouped into genotype IVa of genus Novirhabdovirus. Expression profile of genes in olive flounder was analyzed at day 1 and day3 after infection with this VHSV isolate by using cDNA microarray containing olive flounder 13K cDNA clones. Microarray analysis revealed 785 up-regulated genes and 641 down-regulated genes by at least two-fold in virus-infected fish compared to healthy control groups. Among 785 up-regulated genes, we identified seven immune response-associated genes, including the interferon (IFN)-induced 56-kDa protein (IFI56), suppressor of cytokine signaling 1 (SOCS1), interleukin 8 (IL-8), cluster of differentiation 83 (CD83), α-globin (HBA), VHSV-induced protein-6 (VHSV6), and cluster of differentiation antigen 9 (CD9). Our results confirm previous reports that even virulent strain of VHSV induces expression of genes involved in protective immunity against VHSV. PMID:27599933

  10. Philasterides dicentrarchi, a histophagous ciliate causing scuticociliatosis in olive flounder, Philasterides dicentrarchi--histopathology investigations.

    PubMed

    Harikrishnan, Ramasamy; Jin, Chang-Nam; Kim, Ju-Sang; Balasundaram, Chellam; Heo, Moon-Soo

    2012-03-01

    Philasterides dicentrarchi is a histophagous scuticociliate infecting Korean olive flounder farms in Jeju Island, South Korea, where it causes significant economic losses. However, the route of entry of these parasites in olive flounder is currently unknown. In the present study, we attempted experimental infection with different doses (concentrations) of P. dicentrarchi by intraperitoneal (IP) injection; maximum cumulative mortality of 95% was recorded on the administration of 2.1×10(8) ciliates/ml. In small size group (3 cm length, 24±2 gwt) the ciliates were found in connective tissues of muscle, fins, and nervous tissue while in large size group (5 cm length, 47±3 gwt), the ciliates were predominantly occurred with associated symptoms of liquefaction in the central nervous system. The major clinico-pathological manifestations were loss of scales, appearance of bleached spots that coalesced to form brownish patches, hemorrhagic, and severe dermal necrotic lesion. In addition there was degeneration of muscle fibers, necrosis of the hepatic parenchyma, and severe edema of the intestinal walls, extensive fouling, necrotic degeneration and hyperplasia in the gill bronchial epithelium. In this study, many ciliates were found in the blood vessel, brain, dorsal fins, muscle, kidney, and vertebra of infected flounder. The histopathological investigations are of significant importance in view of possible routes of entry into the host and doses of ciliates that trigger fast infection for potential utility of drugs as a strategy for the control of P. dicentrarchi in farmed olive flounder.

  11. 76 FR 29670 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Quota Transfer

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-23

    ..., Scup, and Black Sea Bass Fishery Management Plan, which was published on December 17, 1993 (58 FR 65936... (76 FR 23206). The difference is 590 lb (268 kg) less of summer flounder to be transferred; therefore... North Carolina quota. Additionally, a vessel experiencing mechanical problems was authorized to land...

  12. Expression Pattern of Early Growth Response Gene 1 during Olive Flounder (Paralichthys olivaceus) Embryonic Development

    PubMed Central

    Yang, Hyun; Lee, Jeong-Ho; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Kim, Kyung-Kil

    2014-01-01

    The early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages. PMID:25949193

  13. Antioxidative responses and bioaccumulation in Japanese flounder larvae and juveniles under chronic mercury exposure.

    PubMed

    Huang, Wei; Cao, Liang; Ye, Zhenjiang; Yin, Xuebo; Dou, Shuozeng

    2010-06-01

    This study investigated the sub-lethal effects of waterborne mercury on growth, bioaccumulation and antioxidative responses of larvae and juveniles of Japanese flounder (Paralichthys olivaceus). Fish were exposed to 0-10 microg Hg(2)(+)L(-1) solutions from embryonic to the juvenile stages for 80 days. Antioxidative responses to mercury exposure were studied in metamorphosing larvae (18 days post hatching, dph), settling larvae (33 dph) and juveniles (78 dph). Results showed that increasing mercury concentration led to increased mercury bioaccumulation and reduced flounder growth. Of the antioxidants investigated, superoxide dismutase (SOD) and catalase (CAT) activities at the three developmental stages were sensitive to mercury exposure and increased with increasing mercury concentration. Glutathione (GSH) content was elevated in metamorphosing larvae, but decreased in juveniles as mercury concentration increased. Glutathione-S-transferase (GST) activity did not significantly vary with mercury concentration in either larvae or juveniles. Mercury exposure did not affect malondialdehyde (MDA) content of larvae, but significantly increased MDA content of juveniles. Results suggest that flounder larvae and juveniles have the potential to manipulate the levels of antioxidants such as SOD, CAT and GSH, which protect flounder from oxidative stress induced by mercury exposure. These antioxidants could serve as biomarkers of mercury contamination in the aquatic environment.

  14. Comparison of antibodies in marine fish from clean and polluted waters of the New York Bight: relative levels against 36 bacteria.

    PubMed Central

    Robohm, R A; Brown, C; Murchelano, R A

    1979-01-01

    Fish from polluted waters are subject to increased prevalence of disease. Because they respond to bacterial pathogens by producing serum antibodies, it was possible to construct a seasonal serological record in three fish species from clean and polluted waters of the New York Bight. Antibody levels were determined by testing sera for agglutinating activity against 36 strains of bacteria. Evaluation of 5,100 antibody titrations showed the following. During warm months, summer flounder (Paralichthys dentatus) from the polluted area had significantly higher antibody levels and antibody to a greater diversity of bacteria than fish from the unpolluted area. Weakfish (Cynoscion regalis) from the same polluted area shared with summer flounder raised titers to many bacteria. The greatest proportion of raised titers was against Vibrio species, although prominent titers were also seen against Aeromonas salmonicida and Haemophilus piscium, bacteria usually associated with diseases in freshwater but not marine fish. Differences between polluted and clean waters were not as evident in winter flounder (Pseudopleuronectes americanus) during cold months. This could be due, in part, to reduced antibody production at colder temperatures. The data illustrate the usefulness of the serum antibody record in identifying environmental exposure to bacteria in marine fish and indicate that the polluted New York Bight apex has increased levels and diversity of bacteria during warm months. PMID:518084

  15. Molecular cloning and expression analysis of the STAT1 gene from olive flounder, Paralichthys olivaceus

    PubMed Central

    Park, Eun-Mi; Kang, Jung-Ha; Seo, Jung Soo; Kim, GunDo; Chung, Jongkyeong; Choi, Tae-Jin

    2008-01-01

    Background Signal transducer and activator of transcription 1 (STAT1) is a critical component of interferon (IFN)-alpha/beta and IFN-gamma signaling. Although seven isoforms of STAT proteins have been reported from mammals, limited information is available for the STAT genes in fish. We isolated complementary DNA with high similarity to mammalian STAT1 from the olive flounder, Paralichthys olivaceus. Results A DNA fragment containing the conserved SH2 domain was amplified by RT-PCR using degenerate primers designed based on the highly conserved sequences in the SH2 domains of the zebrafish and mammalian STAT1. The complete cDNA sequence was obtained by 5' and 3' RACE. The flounder STAT1 transcript consisted of 2,909 bp that encoded a polypeptide of 749 amino acids. The overall similarity between flounder STAT1 and other STATs was very high, with the highest amino acid sequence identity to snakehead (89%). Phylogenetic analyses reveal that flounder STAT1 is in the same monophyletic group with snakehead STAT1. Quantitative real time RT-PCR and in situ hybridization revealed that STAT1 was expressed in almost all examined organs and tissues, with high expression in gill, spleen, kidney, and heart. The accumulation of STAT1 mRNA in different developmental stages, as determined by real time RT-PCR, increased with development. Conclusion Recent cloning of various cytokine genes and the STAT1 gene of olive flounder here suggest that fish also use the highly specialized JAK-STAT pathway for cytokine signaling. Identification of other STAT genes will elucidate in detail the signal transduction system in this fish. PMID:18578892

  16. Stress responses of starry flounder, Platichthys stellatus (Pallas) following water temperature rise.

    PubMed

    Min, Byung Hwa; Park, Mi Seon; Myeong, Jeong-In

    2015-09-01

    Stress responses of starry flounder, Platichthys stellatus (Pallas) following water temperature rise were investigated to establish the influence of ambient temperature on this species. The physiological indicators of stress were plasma cortisol, glucose, aspartate aminotransferase, alanine aminotransferase, sodium, chloride, osmolality and triiodothyronine (T3). No significant difference in plasma parameters were observed among the experimental groups of 15 degrees C, 18 degrees C and 21 degrees C. Level of plasma cortisol (49.0-95.0 ng ml(-1)) and glucose (56.1-58.1 mg dl(-1)) of starry flounders kept at 24 degrees C-27 degrees C were significantly higher than those (cortisol: 20.4-23.6 ng ml(-1), glucose: 40.6-47.1 mg dl(-1)) observed in the 15 degrees C-21 degrees C groups. Changes in aspartate aminotransferase and alanine aminotransferase following water temperature rise showed a similar pattern to plasma cortisol and glucose. Starry flounders exposed to 27 degrees C exhibited higher plasma sodium (164.7 mmol l(-1)), chloride (147.6 mmol l(-1)), and osmolality (450.7 mOsm kg(-1)) than those (sodium: 154.0-158.7 mmol l(-1), chloride: 139.1-140.4 mmol l(-1), osmolality: 375.1-383.8 mOsm kg(-1)) fish exposed to 15-21 degrees C. Though plasma T3 (29.4 ng ml(-1)) of starry flounder increased at 24 degrees C, this hormone was significantly lower (19.3 ng ml(-1)) in fish kept at 27 degrees C than those (24.6 ng ml(-1)) the fish at 15 degrees C. This phenomenon seems to be directly associated with long-term fasting. Accordingly, the results suggested that starry flounders got stressed with osmoregulatory disturbances above 24 degrees C. PMID:26521544

  17. Use of imaging analysis for modeling growth and development of hatchery-reared southern flounder and Florida pompano

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Southern flounder, Paralichthys lethostigma, and Florida pompano, Trachinotus carolinus, are prime candidates for aquaculture. Hatchery methods have been developed for each species, but rearing larvae through metamorphosis is difficult. We investigated the early life history of these fishes using ...

  18. Ocean Acidification Effects on the Early Life-Stages of Commercially Important Flatfish of the Northeast USA

    NASA Astrophysics Data System (ADS)

    Chambers, R. C.; Habeck, E. A.; Candelmo, A. C.; Poach, M.; Wieczorek, D.; Phelan, B.; Caldarone, E.; Cooper, K. R.

    2012-12-01

    The limited available evidence about effects on finfish of high CO2 levels and acidification of our oceans suggests that effects will differ across fish species, be subtle, and interact with other stressors. A carefully planned, experimental framework was developed to cast an extensive yet strategic inferential net. Three key elements of our approach are the use of 1) multiple marine finfish species of relevance to the northeastern USA that differ in their ecologies including spawning season and habitat of early life-stages; 2) a wide yet realistic range of environmental conditions (i.e., concurrent manipulation of CO2 levels and water temperatures), and 3) a diverse set of response variables related to fish sensitivity to elevated CO2 levels, water temperatures, and their interactions. The response variable set reflects fish condition, fitness, and likelihood of recruitment, and includes measures of viability, physiology, histopathology, growth, development, and behavior expressed during fish early life-stages (i.e., gametes, embryos, and larvae). Early life-stages were chosen due to the anticipation of their vulnerability to acid-base challenges in their environment. To date, factorial experiments have been implemented on summer flounder (Paralichthys dentatus) and winter flounder (Pseudopleuronectes americanus). Initial results reveal survival of summer flounder embryos is compromised by pH < 7.7 (CO2 > 790 ppm). These results were similar across offspring groups (i.e., embryos from different parents). Winter flounder are larger at hatching when exposed to high CO2 levels in the coolest environment implemented in our experiments (range 4 to 10 ○C). Further responses of advanced larvae of both flounder species are currently being assessed for evidence of other whole body, component organ, and biochemical impairment. This study will aid researchers and resource managers in identifying species types, life-stages, and biotic responses that are most sensitive to

  19. Consumption of seeds of southwestern white pine (Pinus strobiformis) by Black Bear (Ursus americanus)

    USGS Publications Warehouse

    Mattson, David J.; Arundel, Terry A.

    2013-01-01

    We report a discovery of black bears (Ursus americanus) consuming seeds of southwestern white pine (Pinus strobiformis) on north slopes of the San Francisco Peaks near Flagstaff, Arizona, in high-elevation, mixed-species conifer forest. In one instance, a bear had obtained seeds from cones excavated from a larder horde made by a red squirrel (Tamiasciurus hudsonicus). Consumption of seeds of southwestern white pine by bears had not been previously documented. This discovery adds to the number of species of pine used by bears for food as well as the geographic range within which the behavior occurs.

  20. Mediastinal teratoma in a free-ranging American black bear (Ursus americanus).

    PubMed

    Munk, Brandon A; Turner, J Chris; Keel, M Kevin

    2013-12-01

    A 2.75-yr-old female American black bear (Ursus americanus) was found emaciated and unable to rise. It was euthanized due to the perceived poor prognosis. An approximately 18 x 8 x 10-cm, multiloculated, well-demarcated mass that extended from the bifurcation of the trachea, laterally displacing the lung lobes and caudally displacing the heart. The mass contained fibrous connective tissue, cartilage, bone, nervous tissue, smooth muscle, and a variety of epithelial structures. A mediastinal teratoma was diagnosed based on the microscopic features of the neoplasm. PMID:24450083

  1. SURGICAL CORRECTION OF BILATERAL PATELLAR LUXATION IN AN AMERICAN BLACK BEAR CUB (URSUS AMERICANUS).

    PubMed

    Bennett, Katarina R; Desmarchelier, Marion R; Bailey, Trina R

    2015-06-01

    A wild orphaned male American black bear cub ( Ursus americanus ) presented with hind limb gait abnormalities and was found to have bilateral grade 3 laterally luxating patellas. There were no other significant abnormalities detected on neurologic, radiographic, or hematologic examinations. The trochlear grooves were deepened with a chondroplasty, and the redundant soft tissues imbricated. There was a marked improvement in the bear's gait postoperatively, with an apparent full return to function. To the authors' knowledge, patellar luxation has not been reported in the Ursidae family, and the success in this case suggests that this technique may be used in large wild or captive carnivore cubs. PMID:26056894

  2. Studies on endoparasites of the black bear (Ursus americanus) in the southeastern United States.

    PubMed

    Crum, J M; Nettles, V F; Davidson, W R

    1978-04-01

    Examination of 53 black bears (Ursus americanus) from six states in the southeastern United States revealed at least 17 species of endoparasites, including Sarcocystis sp., Spirometra mansonoides (spargana), Macracanthorhynchus ingens, Ancylostoma caninum, Arthrocephalus lotoris, Baylisascaris transfuga, Capillaria aerophila, Capillaria putorii, Crenosoma sp., Cyathospirura sp., Dirofilaria immitis, Gnathostoma sp., Gongylonema pulchrum, microfilariae, Molineus barbatus, Physaloptera sp. and Strongyloides sp. Twelve of these represent new host records for black bear, and two are considered to be new species. Data are presented on prevalence, intensity and geographic distribution of each species. Pathologic effects were associated with infections of spargana of S. mansonoides and adults of C. aerophilia.

  3. Gene expression responses of European flounder (Platichthys flesus) to 17-beta estradiol.

    PubMed

    Williams, Tim D; Diab, Amer M; George, Stephen G; Sabine, Victoria; Chipman, James K

    2007-02-01

    Male European flounder (Platichthys flesus) were intraperitoneally injected with 10mg/kg 17-beta estradiol and tissues taken from individuals over a timecourse of 16 days. The GENIPOL P. flesus cDNA microarray was employed to detect hepatic gene expression differences between fish treated with estradiol and saline controls. Known biomarkers of estrogen exposure, choriogenin L and vitellogenins, showed sustained induction over the time-course. Among 175 identified clones showing sustained statistically significant induction or repression, those associated with the Gene Ontology terms mitochondria, amino acid synthesis, ubiquitination and apoptosis were included amongst those induced while those associated with immune function, electron transport, cell signalling and protein phosphorylation were repressed. Thus, we show the gene expression response of an environmentally relevant fish species to a high dose of an estrogenic endocrine disruptor and also report the sequencing of a further 2121 flounder ESTs.

  4. Does the chronic chemical contamination of a European flounder population decrease its thermal tolerance?

    PubMed

    Lavergne, Edouard; Pedron, Nicolas; Calves, Isabelle; Claireaux, Guy; Mazurais, David; Zambonino-Infante, José; Le Bayon, Nicolas; Cahu, Chantal; Laroche, Jean

    2015-06-30

    Juvenile flounders (Platichthys flesus), collected in two estuaries with similar temperature regimes (the heavily polluted Seine and the moderately contaminated Vilaine), were submitted to a common garden experiment. After an acclimation period, both populations were challenged by a thermal stress (9-24°C for 15days, then maintenance at 24°C for 19days). The condition factor of the Vilaine fish increased in both conditions, while it decreased for the heated Seine flounders after 34days. The expression of genes related to the energetic metabolism was measured in the liver. The expression levels for ATP-F0 and COII were significantly reduced for heated vs. standard fish from both estuaries, while a decrease of the 12S expression was detected only in heated vs. standard fish from the Seine estuary. Thus, it is suggested that highly contaminated fish from Seine could display a lower tolerance to thermal stress, compared to moderately contaminated fish from Vilaine.

  5. Cloning and expression of prion protein encoding gene of flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Zhang, Zhiwen; Sun, Xiuqin; Zhang, Jinxing; Zan, Jindong

    2008-02-01

    The prion protein (PrP) encoding gene of flounder ( Paralichthys olivaceus) was cloned. It was not interrupted by an intron. This gene has two promoters in its 5' upstream, indicating that its transcription may be intensive, and should have an important function. It was expressed in all 14 tissues tested, demonstrating that it is a house-keeping gene. Its expression in digestion and reproduction systems implies that the possible prions of fish may transfer horizontally.

  6. Dynamics of summer flounder, Paralichthys dentatus, seasonal migrations based on ultrasonic telemetry

    NASA Astrophysics Data System (ADS)

    Sackett, Dana K.; Able, Kenneth W.; Grothues, Thomas M.

    2007-08-01

    Migrations of summer flounder, Paralichthys dentatus, to and from estuaries to the continental shelf in the Mid-Atlantic Bight (MAB) occur seasonally but their dynamics are poorly understood. Ultrasonic telemetry, both passive and active, was used during 2003-2005 to determine timing and rate of juvenile and adult summer flounder (268-535 mm TL) migrating to and from the Mullica River-Great Bay estuary in southern New Jersey. Additionally, 7 years of inner continental shelf surveys off New Jersey were used to assess complementary seasonal movements. Most tagged fish emigrated from the estuary between July and September, though emigration lasted into December and appeared to be influenced by a number of factors. In July 2004, more tagged fish emigrated, at increased rates of movement, at low barometric pressure during a storm event. Trawl collections on the inner shelf demonstrated the same approximate immigration times as seen with telemetry. Later in the fall, increased numbers of tagged summer flounder emigrated from the estuary when dissolved oxygen was decreasing. Fall trawl surveys showed increased numbers of fish on the inner shelf when dissolved oxygen was decreasing in the Mullica River-Great Bay estuary, supporting the telemetry results. Fish emigrated from the estuary during the day and night but nighttime movements were in deeper water at slightly slower rates of movement. Exit and re-entry also occurred during the fall emigration. Ultrasonically tagged individuals demonstrated homing by returning to the same estuary, in March through June, in the second and third year of the study (39-6%, respectively). In summary, immigration may result from homing for a large proportion of summer flounder. Emigration may be associated with storms on an episodic scale, and dissolved oxygen and temperature on a seasonal scale.

  7. 78 FR 76766 - Fisheries of the Northeastern United States; Summer Flounder Fishery; Commercial Quota Harvested...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-19

    ... is 11,793,596 lb (5,349,575 kg) (77 FR 76942, December 31, 2012). The percent allocated to vessels... Fisheries, NOAA (AA), finds good cause pursuant to 5 U.S.C. 553(b)(B) to waive prior notice and the... Summer Flounder Fishery Management Plan. The AA further finds, pursuant to 5 U.S.C. 553(d)(3), good...

  8. Ontogenetic changes in digestive enzyme activities and the amino acid profile of starry flounder Platichthys stellatus

    NASA Astrophysics Data System (ADS)

    Song, Zhidong; Wang, Jiying; Qiao, Hongjin; Li, Peiyu; Zhang, Limin; Xia, Bin

    2016-09-01

    Ontogenetic changes in digestive enzyme activities and the amino acid (AA) profile of starry flounder, Platichthys stellatus, were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements. Larvae were collected at the egg stage and 0, 2, 4, 7, 12, 17, 24 days after hatching (DAH) for analysis. Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH. Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH. Pepsin activity increased sharply beginning 17 DAH. Lipase activity increased significantly 4 DAH and increased progressively with larval growth. Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease. High free amino acid (FAA) content was detected in starry flounder eggs (110.72 mg/g dry weight). Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae. Most FAAs (except lysine and methionine) decreased >50% in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae. Changes in the protein amino acid (PAA) profile were much milder than those observed for FAAs. Most PAAs increased gradually during larval development, except lysine and phenylalanine. The percentages of free threonine, valine, isoleucine, and leucine decreased until the end of the trial, whereas the protein forms of these four AAs followed the opposite trend. A comparison of the essential AA composition of live food (rotifers, Artemia nauplii, and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA. These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food. Moreover, the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed.

  9. Evaluating the Hypoxia Response of Ruffe and Flounder Gills by a Combined Proteome and Transcriptome Approach

    PubMed Central

    Tiedke, Jessica; Borner, Janus; Beeck, Hendrik; Kwiatkowski, Marcel; Schmidt, Hanno; Thiel, Ralf; Fabrizius, Andrej; Burmester, Thorsten

    2015-01-01

    Hypoxia has gained ecological importance during the last decades, and it is the most dramatically increasing environmental factor in coastal areas and estuaries. The gills of fish are the prime target of hypoxia and other stresses. Here we have studied the impact of the exposure to hypoxia (1.5 mg O2/l for 48 h) on the protein expression of the gills of two estuarine fish species, the ruffe (Gymnocephalus cernua) and the European flounder (Platichthys flesus). First, we obtained the transcriptomes of mixed tissues (gills, heart and brain) from both species by Illumina next-generation sequencing. Then, the gill proteomes were investigated using two-dimensional gel electrophoresis and mass spectrometry. Quantification of the normalized proteome maps resulted in a total of 148 spots in the ruffe, of which 28 (18.8%) were significantly regulated (> 1.5-fold). In the flounder, 121 spots were found, of which 27 (22.3%) proteins were significantly regulated. The transcriptomes were used for the identification of these proteins, which was successful for 15 proteins of the ruffe and 14 of the flounder. The ruffe transcriptome dataset comprised 87,169,850 reads, resulting in an assembly of 72,108 contigs (N50 = 1,828 bp). 20,860 contigs (26.93%) had blastx hits with E < 1e-5 in the human sequences in the RefSeq database, representing 14,771 unique accession numbers. The flounder transcriptome with 78,943,030 reads assembled into 49,241 contigs (N50 = 2,106 bp). 20,127 contigs (40.87%) had a hit with human proteins, corresponding to 14,455 unique accession numbers. The regulation of selected genes was confirmed by quantitative real-time RT-PCR. Most of the regulated proteins that were identified by this approach function in the energy metabolism, while others are involved in the immune response, cell signalling and the cytoskeleton. PMID:26273839

  10. Evaluating the Hypoxia Response of Ruffe and Flounder Gills by a Combined Proteome and Transcriptome Approach.

    PubMed

    Tiedke, Jessica; Borner, Janus; Beeck, Hendrik; Kwiatkowski, Marcel; Schmidt, Hanno; Thiel, Ralf; Fabrizius, Andrej; Burmester, Thorsten

    2015-01-01

    Hypoxia has gained ecological importance during the last decades, and it is the most dramatically increasing environmental factor in coastal areas and estuaries. The gills of fish are the prime target of hypoxia and other stresses. Here we have studied the impact of the exposure to hypoxia (1.5 mg O2/l for 48 h) on the protein expression of the gills of two estuarine fish species, the ruffe (Gymnocephalus cernua) and the European flounder (Platichthys flesus). First, we obtained the transcriptomes of mixed tissues (gills, heart and brain) from both species by Illumina next-generation sequencing. Then, the gill proteomes were investigated using two-dimensional gel electrophoresis and mass spectrometry. Quantification of the normalized proteome maps resulted in a total of 148 spots in the ruffe, of which 28 (18.8%) were significantly regulated (> 1.5-fold). In the flounder, 121 spots were found, of which 27 (22.3%) proteins were significantly regulated. The transcriptomes were used for the identification of these proteins, which was successful for 15 proteins of the ruffe and 14 of the flounder. The ruffe transcriptome dataset comprised 87,169,850 reads, resulting in an assembly of 72,108 contigs (N50 = 1,828 bp). 20,860 contigs (26.93%) had blastx hits with E < 1e-5 in the human sequences in the RefSeq database, representing 14,771 unique accession numbers. The flounder transcriptome with 78,943,030 reads assembled into 49,241 contigs (N50 = 2,106 bp). 20,127 contigs (40.87%) had a hit with human proteins, corresponding to 14,455 unique accession numbers. The regulation of selected genes was confirmed by quantitative real-time RT-PCR. Most of the regulated proteins that were identified by this approach function in the energy metabolism, while others are involved in the immune response, cell signalling and the cytoskeleton. PMID:26273839

  11. Molecular cloning and characterization of interferon regulatory factor 9 (IRF9) in Japanese flounder, Paralichthys olivaceus.

    PubMed

    Hu, Guo-Bin; Zhao, Ming-Yu; Lin, Jing-Yun; Liu, Qiu-Ming; Zhang, Shi-Cui

    2014-08-01

    Interferon regulatory factor 9 (IRF9) in mammals is known to be involved in antiviral response. In this study, we studied the structure, mRNA tissue distribution and regulation of IRF9 from Japanese flounder, Paralichthys olivaceus. The cDNA sequence of IRF9 is 3305 bp long, containing an open reading frame (ORF) of 1308 bp that encodes a peptide of 435 amino acids. The predicted protein sequence shares 33.7-72.0% identity to other fish IRF9s. Japanese flounder IRF9 possesses a DNA-binding domain (DBD), an IRF association domain (IAD), two nuclear localization signals (NLSs) and a proline-rich domain (PRD). The IRF9 transcripts were detectable in all examined tissues of healthy Japanese flounders, with higher levels in the head kidney, kidney, liver and spleen. The IRF9 mRNA levels were up-regulated in the gills, head kidney, spleen and muscle when challenged with polyinosinic:polycytidylic acid (poly I:C) or lymphocystis disease virus (LCDV). The up-regulations were stronger and arose earlier in the case of poly I:C treatment in most tested organs in a 7-day time course, with maximum increases ranging from 1.37- to 8.59-fold and peak time points from 3 h to 3 d post injection depending on different organs, relative to those in the case of LCDV treatment which ranged from 1.32- to 3.21-fold and from 18 h to 3 d post injection, respectively. The highest and earliest inductions were detected in the spleen in both challenge cases, while the inductions by LCDV in the muscle were quite faint. These results demonstrate a role of Japanese flounder IRF9 in the host's antiviral responses. PMID:24837327

  12. VITELLOGENIN-INDUCED PATHOLOGY AND MORTALITY IN MALE SUMMER FLOUNDER (PARALICHTHYS DENTATUS)

    EPA Science Inventory

    Male summer flounder (Paralichthys dentatus) were given two injections (initially and two weeks later) of 17 b-estradiol (E2) totaling 0.2 (2 x 0.1), 2.0 (2 x 1.0) or 20.0 (2 x 10.0) mg E2/kg body weight. Blood and tissue samples were collected 4, 6 and 8 weeks after the initial ...

  13. Sarcocystis sp. in mountain goats (Oreamnos americanus) in Washington: prevalence and search for the definitive host.

    PubMed

    Foreyt, W J

    1989-10-01

    During October and November 1986, Sarcocystis sp. was detected in 24 of 56 (43%) tongues from hunter-killed mountain goats (Oreamnos americanus) in Washington (USA). Sarcocysts had an unusual sessile polyp-shaped branched wall. Mean size of 154 sarcocysts was 71.3 x 37.8 microns (range, 20 to 248 x 10 to 120 microns), and the mean intensity was 2.3 (range, 1 to 28). In an attempt to identify the definitive host, infected tongues were fed to four coyotes (Canis latrans), eight domestic dogs, four domestic cats, three bears (Ursus americanus), two raccoons (Procyon lotor), two martens (Martes americana), two fishers (Martes pennanti), three skunks (Mephitis mephitis), five mink (Mustela vison), five ferrets (Mustela putorius), one pigtail macaque (Macaca nemestrina), two red-tailed hawks (Buteo jamaicensis) and one great horned owl (Bubo virginianus). Oocysts or sporocysts were not detected in the feces of any host for less than or equal to 20 days after ingestion of the infected meat. The definitive host for Sarcocystis sp. in mountain goats was not identified.

  14. Seasonal and daily plasma corticosterone rhythms in American toads, Bufo americanus

    SciTech Connect

    Pancak, M.K.; Taylor, D.H.

    1983-06-01

    Concentrations of corticosterone were measured in the plasma of American toads, Bufo americanus, on a seasonal basis using a radioimmunoassay technique. Two populations of toads, maintained under different light conditions, were monitored to observe the effects of photoperiod on the seasonal rhythm of plasma corticosterone. Under a natural photoperiod toads demonstrated a rhythm consisting of a spring peak and a fall peak in corticosterone concentration. Toads maintained under a 12L:12D photoperiod all year round demonstrated a similar rhythm with peaks in the spring and fall. This suggests that an endogenous (circannual) rhythm of corticosterone may be playing an important role in the seasonal change of overt behavior and physiology of Bufo americanus. A daily rhythm of corticosterone was also detected in toads when blood samples were taken every 4 hr. When compared to a previously published circadian rhythm study of locomotor activity, the surge in corticosterone concentration for the day occurred at 1730 just prior to the peak in locomotor activity.

  15. Aspartic cathepsin D endopeptidase contributes to extracellular digestion in clawed lobsters Homarus americanus and Homarus gammarus.

    PubMed

    Rojo, Liliana; Muhlia-Almazan, Adriana; Saborowski, Reinhard; García-Carreño, Fernando

    2010-11-01

    Acid digestive proteinases were studied in the gastric fluids of two species of clawed lobster (Homarus americanus and Homarus gammarus). An active protein was identified in both species as aspartic proteinase by specific inhibition with pepstatin A. It was confirmed as cathepsin D by mass mapping, N-terminal, and full-length cDNA sequencing. Both lobster species transcribed two cathepsin D mRNAs: cathepsin D1 and cathepsin D2. Cathepsin D1 mRNA was detected only in the midgut gland, suggesting its function as a digestive enzyme. Cathepsin D2 mRNA was found in the midgut gland, gonads, and muscle. The deduced amino acid sequence of cathepsin D1 and cathepsin D2 possesses two catalytic DTG active-site motifs, the hallmark of aspartic proteinases. The putatively active cathepsin D1 has a molecular mass of 36.4 kDa and a calculated pI of 4.14 and possesses three potential glycosylation sites. The sequences showed highest similarities with cathepsin D from insects but also with another crustacean cathepsin D. Cathepsin D1 transcripts were quantified during a starvation period using real-time qPCR. In H. americanus, 15 days of starvation did not cause significant changes, but subsequent feeding caused a 2.5-fold increase. In H. gammarus, starvation caused a 40% reduction in cathepsin D1 mRNA, and no effect was observed with subsequent feeding. PMID:20169386

  16. Comparative tissue expression of American lobster (Homarus americanus) immune genes during bacterial and scuticociliate challenge.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Wang, Haili; Greenwood, Spencer J

    2015-12-01

    The American lobster (Homarus americanus) fishery is the most economically significant fishery in Canada; although comparatively little is known about the lobsters' response to pathogenic challenge. This is the first study to investigate the expression of immune genes in tissues outside of the lobster hepatopancreas in response to challenges by the Gram-positive bacteria, Aerococcus viridans var. homari or the scuticociliate parasite, Anophryoides haemophila. The hepatopancreas has been regarded as the major humoral immune organ in crustaceans, but the contribution of other organs and tissues to the molecular immune response has largely been overlooked. This study used RT-qPCR to monitor the gene expression of several immune genes including three anti-lipopolysaccharide isoforms (ALF) Homame ALF-B1, Homame ALF-C1 and ALFHa-1, acute phase serum amyloid protein A (SAA), as well as thioredoxin and hexokinase, in antennal gland and gill tissues. Our findings indicate that the gene expression of the SAA and all ALF isoforms in the antennal gland and gill tissues increased in response to pathogenic challenge. However, there was differential expression of individual ALF isoforms that were dependent on both the tissue, and the pathogen used in the challenge. The gene expression changes of several immune genes were found to be higher in the antennal gland than have been previously reported for the hepatopancreas. This study demonstrates that increased immune gene expression from the gill and antennal gland over the course of pathogen induced disease contributes to the immune response of H. americanus.

  17. Cloning and expression study of the lobster (Homarus americanus) vitellogenin: Conservation in gene structure among decapods.

    PubMed

    Tiu, Shirley Hiu Kwan; Hui, Ho-Lam; Tsukimura, Brian; Tobe, Stephen S; He, Jian-Guo; Chan, Siu-Ming

    2009-01-01

    This study reports the molecular characterization of the vitellogenin (Vg) of the lobster, Homarus americanus. Based on the annual collection of female lobsters, vitellogenesis commences in early March and continues through to September of each year. Using an antibody to vitellin of the lobster, H. americanus, several immunoreactive ovarian proteins were initially identified by Western blot analysis. The 80kDa protein contained the amino acid sequence APWGGNTPRC, identified subsequently by cDNA cloning to be identical to the lobster Vg. In common with the shrimp Metapenaeus ensis and crab Charybdis feriatus, the lobster HaVg1 gene comprises 14 introns and 15 exons. The deduced HaVg1 precursor is most similar to the Vg of the crayfish Cherax quadricarinatus (57%), followed by M. ensis (40-43% identity) and C. feriatus (38%). The results from genomic and RT-PCR cloning also confirmed the presence of multiple Vg genes in lobster. At early reproductive stages, the hepatopancreas HaVg1 transcript levels are low but increased to a maximum in animals with mature oocytes. The ovary, however, also expressed low levels of HaVg1. Using in vitro explant culture, treatment of hepatopancreas fragments with farnesoic acid or 20-hydroxyecdysone resulted in a significant stimulation in HaVg1 expression. From this study, it appears that Vg gene organization and expression pattern in decapods is highly conserved. Similar endocrine mechanisms may govern the process of vitellogenesis across the decapods.

  18. Sarcocystis sp. in mountain goats (Oreamnos americanus) in Washington: prevalence and search for the definitive host.

    PubMed

    Foreyt, W J

    1989-10-01

    During October and November 1986, Sarcocystis sp. was detected in 24 of 56 (43%) tongues from hunter-killed mountain goats (Oreamnos americanus) in Washington (USA). Sarcocysts had an unusual sessile polyp-shaped branched wall. Mean size of 154 sarcocysts was 71.3 x 37.8 microns (range, 20 to 248 x 10 to 120 microns), and the mean intensity was 2.3 (range, 1 to 28). In an attempt to identify the definitive host, infected tongues were fed to four coyotes (Canis latrans), eight domestic dogs, four domestic cats, three bears (Ursus americanus), two raccoons (Procyon lotor), two martens (Martes americana), two fishers (Martes pennanti), three skunks (Mephitis mephitis), five mink (Mustela vison), five ferrets (Mustela putorius), one pigtail macaque (Macaca nemestrina), two red-tailed hawks (Buteo jamaicensis) and one great horned owl (Bubo virginianus). Oocysts or sporocysts were not detected in the feces of any host for less than or equal to 20 days after ingestion of the infected meat. The definitive host for Sarcocystis sp. in mountain goats was not identified. PMID:2509738

  19. Comparative tissue expression of American lobster (Homarus americanus) immune genes during bacterial and scuticociliate challenge.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Wang, Haili; Greenwood, Spencer J

    2015-12-01

    The American lobster (Homarus americanus) fishery is the most economically significant fishery in Canada; although comparatively little is known about the lobsters' response to pathogenic challenge. This is the first study to investigate the expression of immune genes in tissues outside of the lobster hepatopancreas in response to challenges by the Gram-positive bacteria, Aerococcus viridans var. homari or the scuticociliate parasite, Anophryoides haemophila. The hepatopancreas has been regarded as the major humoral immune organ in crustaceans, but the contribution of other organs and tissues to the molecular immune response has largely been overlooked. This study used RT-qPCR to monitor the gene expression of several immune genes including three anti-lipopolysaccharide isoforms (ALF) Homame ALF-B1, Homame ALF-C1 and ALFHa-1, acute phase serum amyloid protein A (SAA), as well as thioredoxin and hexokinase, in antennal gland and gill tissues. Our findings indicate that the gene expression of the SAA and all ALF isoforms in the antennal gland and gill tissues increased in response to pathogenic challenge. However, there was differential expression of individual ALF isoforms that were dependent on both the tissue, and the pathogen used in the challenge. The gene expression changes of several immune genes were found to be higher in the antennal gland than have been previously reported for the hepatopancreas. This study demonstrates that increased immune gene expression from the gill and antennal gland over the course of pathogen induced disease contributes to the immune response of H. americanus. PMID:26551049

  20. Effects of ocean acidification on the larval growth of olive flounder (Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Kim, K.-S.; Shim, J. H.; Kim, S.

    2013-04-01

    Little is known about how marine fishes respond to the reduced pH condition caused by the increased CO2 in the atmosphere. We investigated the effects of CO2 concentration on the growth of olive flounder (Paralichthys olivaceus) larvae. Newly hatched larvae were reared in three different concentrations of CO2 (574, 988 and 1297 μatm CO2) in temperature-controlled water tanks until metamorphosis (4 weeks). Body lengths, weights, and the concentration of some chemical elements in larval tissue were measured at the completion of each experiment, and experiment was repeated three times in May, June, and July 2011. Results indicated that body length and weight of flounder larvae were significantly increased with increasing CO2 concentration (P < 0.05). Daily growth rates of flounder larvae were higher (0.391 mm) from the high CO2 concentration (1297 μatm) than those (0.361 mm and 0.360 mm) from the lower ones (988 and 574 μatm).The measurement on some chemical elements (Ca, Fe, Cu, Zn and Sr) in fish tissue also revealed the increasing tendency of element concentration with increasing CO2 in seawater, although statistical significance cannot be tested due to the single measurement. It suggests that there are enrichment processes of these cations in larval tissue in the low pH condition.

  1. Comparison of proteome typing and serotyping of Streptococcus parauberis isolates from olive flounder (Paralichthys olivaceus).

    PubMed

    Kim, Si Won; Jang, Ho Bin; Lee, Jung Seok; Im, Se Pyeong; Lazarte, Jassy Mary S; Seo, Jong Pyo; Lee, Woo Jai; Kim, Jae Sung; Jung, Tae Sung

    2015-11-01

    The olive flounder (Paralichthys olivaceus) is a cultivated marine species that is economically important in Korea and Japan. Several bacterial pathogens have caused severe mortalities in farmed olive flounder, especially Streptococcus parauberis. We collected 145 S. parauberis isolates from diseased olive flounders from 2003 to 2008 in Jeju Island, South Korea and characterized them by Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS) and by serology. The serological analysis divided the isolates into serotype I (62.1%) and serotype II (36.6%) and the proteome analysis divided the isolates into cluster 1 (43.4%) and cluster 2 (56.6%). All cluster 1 isolates had serotype I, but cluster 2 consisted of serotype I (32.9%), serotype II (64.6%), and others (2.5%). Further detailed analysis of the mass spectra led to identification of several specific m/z peaks that enabled discrimination between cluster 1 and 2 and between serotype I and II within cluster 2. Our results suggest that MALDI TOF MS analysis has potential as an alternative method for the rapid and reliable identification of the fish pathogen S. parauberis.

  2. Species profiles: Life histories and environmental requirements of coastal fishes and invertebrates (North Atlantic): Winter flounder

    SciTech Connect

    Buckley, J.

    1989-01-01

    Species profiles are literature summaries of the taxonomy, life history, and environmental requirements of coastal fishes and aquatic invertebrates. They are designed to assist with environmental impact assessments. From 1935 to 1980, the annual commercial landings of winter flounder in New England ranged from 6000 to 15000 t; the sport catch exceeded the commercial catch in some years. Winter flounder are found in waters with temperatures of 0 to 25 /degree/C and they usually spawn at 0 to 3 /degree/C. Reported fecundities are 0.5 to 1.5 million eggs per female. Metamorphosis from larva to juvenile is complete in 49-80 days, depending on temperature. Juveniles remain in or near shallow natal waters for much of their first 2 years. Adults of some populations move more than 3 miles offshore to cooler waters in summer. These adults live in shallow inshore waters in winter and early spring. Adult winter flounder feed largely on annelids, cnidariids, and mollusks. 58 refs., 2 figs., 1 tab.

  3. Single nucleotide polymorphism of FSHβ gene associated with reproductive traits in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    He, Feng; Wen, Haishen; Yu, Dahui; Li, Jifang; Shi, Bao; Chen, Caifang; Zhang, Jiaren; Jin, Guoxiong; Chen, Xiaoyan; Shi, Dan; Yang, Yanping

    2010-12-01

    Follicle stimulating hormone β (FSHβ) of Japanese flounder ( Paralichthys olivaceus) plays a key role in the regulation of gonadal development. This study aimed to investigate molecular genetic characteristics of the FSHβ gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of FSHβ on reproductive traits in Japanese flounder. We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the FSHβ gene in 60 individuals. We identified only an SNP (T/C) in the coding region of exon3 of FSHβ. The SNP (T/C) did not lead to amino acid changes at the position 340 bp of FSHβ gene. Statistical analysis showed that the SNP was significantly associated with testosterone (T) level and gonadosomatic index (GSI) ( P < 0.05). Individuals with genotype TC of the SNP had significantly higher serum T levels and GSI ( P < 0.05) than that of genotype CC. Therefore, FSHβ gene could be a useful molecular marker in selection for prominent reproductive trait in Japanese Flounder.

  4. Expression Analysis of Cathepsin F during Embryogenesis and Early Developmental Stage in Olive Flounder (Paralichthys olivaceus)

    PubMed Central

    Lee, Jang-Wook; Lee, Young Mee; Yang, Hyun; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Hwang, In Joon; Kim, Sung Yeon; Lee, Jeong-Ho

    2013-01-01

    Cathepsins are members of the multigene family of lysosomal cysteine proteinases and have regulated function in several life processes. The potential role of cathepsin F cysteine gene was expected as protease in the yolk processing mechanism during early developmental stage, but expression analysis was unknown after fertilization. The alignment analysis showed that amino acid sequence of cathepsin F from olive flounder liver expressed sequence tag (EST) homologous to cathepsin F of other known cathepsin F sequences with 87-98% identity. In this study, we examined the gene expression analysis of cathepsin F in various tissues at variety age flounder. Tissue distribution of the cathepsin F mRNA has been shown to be ubiquitous and constitutive pattern regardless of age in each group, although derived from cDNA library using liver sample. The mRNA level of cathepsin F more increased as developmental proceed during embryogenesis and early developmental stage, especially increased in the blastula, hatching stage and 3 days post hatching (dph). As a result, it may suggest that the proteolysis of yolk proteins (YPs) has been implicated as a mechanism for nutrient supply during early larval stages in olive flounder. PMID:25949137

  5. Helminth community structure of sympatric eastern American toad, Bufo americanus americanus, northern leopard frog, Rana pipiens, and blue-spotted salamander, Ambystoma laterale, from southeastern Wisconsin.

    PubMed

    Bolek, Matthew G; Coggins, James R

    2003-08-01

    One-hundred twelve amphibians, including 51 blue-spotted salamanders, Ambystoma laterale, 30 eastern American toads, Bufo americanus americanus, and 31 northern leopard frogs, Rana pipiens, were collected during April-October 1996 from Waukesha County, Wisconsin and examined for helminth parasites. The helminth compound community of this amphibian assemblage consisted of at least 10 species: 9 in American toads, 8 in leopard frogs, and 3 in blue-spotted salamanders. American toads shared 7 species with leopard frogs, and 2 species occurred in all 3 host species. Although there was a high degree of helminth species overlap among these sympatric amphibians, statistically significant differences were found among host species and percent of indirect or direct-life cycle parasites of amphibian species individual component communities (chi2 = 1,015, P < 0.001). American toads had a higher relative abundance of nematodes, 59%, than larval cestodes, 31%, and larval and adult trematodes, 10%, whereas leopard frogs had a higher relative abundance of larval cestodes, 71.3%, and larval and adult trematodes, 25.3%, than nematodes 3.4%. This is related to ecological differences in habitat and dietary preferences between these 2 anuran species. Helminth communities of blue-spotted salamanders were depauperate and were dominated by larval trematodes, 94%, and few nematodes, 6%. Low helminth species richness in this host species is related to this salamander's relatively small host body size, smaller gape size, lower vagility, and more fossorial habitat preference than the other 2 anuran species. Adult leopard frogs and toads had significantly higher mean helminth species richness than metamorphs, but there was no significant difference in mean helminth species richness among adult and metamorph blue-spotted salamanders. Considering adult helminths, the low species richness and low vagility of caudatans as compared with anurans suggest that local factors may be more important in

  6. Isolation and characterization of new genetic types of toxoplasma gondii and prevalence of trichinella murrelli from black bear (Ursus americanus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Black bears (Ursus americanus) are hosts for two important zoonotic parasites, Toxoplasma gondii and Trichinella spp. and bears are hunted for human consumption in the USA. Little is known of the genetic diversity of T. gondii circulating in wildlife. In the present study, antibodies to T. gondii we...

  7. The hidden history of the snowshoe hare, Lepus americanus: extensive mitochondrial DNA introgression inferred from multilocus genetic variation.

    PubMed

    Melo-Ferreira, José; Seixas, Fernando A; Cheng, Ellen; Mills, L Scott; Alves, Paulo C

    2014-09-01

    Hybridization drives the evolutionary trajectory of many species or local populations, and assessing the geographic extent and genetic impact of interspecific gene flow may provide invaluable clues to understand population divergence or the adaptive relevance of admixture. In North America, hares (Lepus spp.) are key species for ecosystem dynamics and their evolutionary history may have been affected by hybridization. Here we reconstructed the speciation history of the three most widespread hares in North America - the snowshoe hare (Lepus americanus), the white-tailed jackrabbit (L. townsendii) and the black-tailed jackrabbit (L. californicus) - by analysing sequence variation at eight nuclear markers and one mitochondrial DNA (mtDNA) locus (6240 bp; 94 specimens). A multilocus-multispecies coalescent-based phylogeny suggests that L. americanus diverged ~2.7 Ma and that L. californicus and L. townsendii split more recently (~1.2 Ma). Within L. americanus, a deep history of cryptic divergence (~2.0 Ma) was inferred, which coincides with major speciation events in other North American species. While the isolation-with-migration model suggested that nuclear gene flow was generally rare or absent among species or major genetic groups, coalescent simulations of mtDNA divergence revealed historical mtDNA introgression from L. californicus into the Pacific Northwest populations of L. americanus. This finding marks a history of past reticulation between these species, which may have affected other parts of the genome and influence the adaptive potential of hares during climate change.

  8. The effects of combined dietary probiotics Lactococcus lactis BFE920 and Lactobacillus plantarum FGL0001 on innate immunity and disease resistance in olive flounder (Paralichthys olivaceus).

    PubMed

    Beck, Bo Ram; Kim, Daniel; Jeon, Jongsu; Lee, Sun-Min; Kim, Hui Kwon; Kim, Oi-Jin; Lee, Jae Il; Suh, Byung Sun; Do, Hyung Ki; Lee, Kwan Hee; Holzapfel, Wilhelm H; Hwang, Jee Youn; Kwon, Mun Gyeong; Song, Seong Kyu

    2015-01-01

    The effects of a dietary probiotic mixture containing Lactococcus (Lc.) lactis BFE920 isolated from bean sprout and autochthonous Lactobacillus (Lb.) plantarum FGL0001 originally isolated from the hindgut of olive flounder (Paralichthys olivaceus) were investigated for the purpose of improving the probiotic effects of Lc. lactis BFE920 on the olive flounder. The immunostimulatory, disease protective, and weight gain effects of Lc. lactis BFE920 were significantly improved when olive flounder (average weight 37.5±1.26 g) were fed the probiotic mixture (log10 7.0 CFU each/g feed pellet) for 30 days. Flounder fed the mixture showed improved skin mucus lysozyme activity and phagocytic activity of innate immune cells compared to flounder fed a single probiotic agent or a control diet. While the levels of neutrophil activity in flounder fed the single probiotic agent or the mixture were similar, they were significantly higher than levels in a control group. Additionally, probiotic-fed flounder showed significantly increased expressions of IL-6, IL-8, and TNF-α in the intestine compared to the control group. Following a 30-day period of being fed probiotics or a control diet, the olive flounder were challenged with an i.p. injection of Streptococcus iniae (log10 6.0 CFU/fish). The groups fed the mixed probiotics, Lc. lactis BFE920, Lb. plantarum FGL0001, and the control diet had survival rates of 55%, 45%, 35%, and 20%, respectively. Flounder fed the probiotic mixture gained 38.1±2.8% more body weight compared to flounder fed the control diet during the 30-day study period. These data strongly suggest that a mixture of Lc. lactis BFE920 and Lb. plantarum FGL0001 may serve as an immunostimulating feed additive useful for disease protection in the fish farming industry. PMID:25449382

  9. Assessment of the European flounder responses to chemical stress in the English Channel, considering biomarkers and life history traits.

    PubMed

    Dupuy, Célie; Galland, Claire; Pichereau, Vianney; Sanchez, Wilfried; Riso, Ricardo; Labonne, Maylis; Amara, Rachid; Charrier, Grégory; Fournier, Michel; Laroche, Jean

    2015-06-30

    A multi-biomarker approach was developed to evaluate responses of European flounder (Platichthys flesus) in three contrasted estuaries over the English Channel: the Canche (pristine site), Tamar (heavy metals and PAHs contamination) and Seine (heavily pollution with a complex cocktail of contaminants). The condition factor and several biomarkers of the immune system, antioxidant enzymes, energetic metabolism and detoxification processes were investigated in young-of-the-year (0+) and one-year-old (1+) flounder. Results underlined the difference between the pristine site and the Seine estuary which showed a lower condition factor, a modulation of the immune system, a higher Cytochrome C oxidase activity, and an up-regulation of BHMT expression. The moderate biomarker responses in the Tamar fish could be linked to the specific contamination context of this estuary. Flounder life history traits were analyzed by otolith microchemistry, in order to depict how the fish use their habitat and thus respond to chemical stress in estuaries.

  10. Comparison of common persistent organic pollutants (POPs) in flounder (Platichthys flesus) from the Vistula (Poland) and Douro (Portugal) River estuaries.

    PubMed

    Waszak, Ilona; Dabrowska, Henryka; Komar-Szymczak, Katarzyna

    2014-04-15

    Groups of flounder (Platichthys flesus) females were collected in 2011 from the Vistula River and the Duoro River estuaries and corresponding reference sites in the southern Baltic Sea and Portuguese coast of the Atlantic Ocean to measure and compare the levels and profiles of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), and polybrominated diphenyl ethers (PBDEs). The estuaries' sediments were also investigated. Several differences were found in the POPs between the estuaries and between the two marine regions, which were highlighted by PCA. The Vistula River estuary POPs, significantly higher than in the Douro River estuary, were dominated by DDTs followed by PCBs. PBDEs levels, indifferent between the estuaries, were relatively low. The POP levels in flounder and sediment evaluated against environmental assessment criteria (EACs) indicated that none of the measured contaminants for which EAC had been established exceeded the criterion, except for CB-118 in flounder from the Vistula River estuary.

  11. 'Candidatus Liberibacter americanus', associated with citrus huanglongbing (greening disease) in São Paulo State, Brazil.

    PubMed

    Teixeira, Diva do Carmo; Saillard, Colette; Eveillard, Sandrine; Danet, Jean Luc; da Costa, Paulo Inácio; Ayres, Antonio Juliano; Bové, Joseph

    2005-09-01

    , were found to have identical or almost identical sequences. In contrast, the intergenic regions of the SPS-HLB liberibacter, 'Ca. L. asiaticus' and 'Ca. L. africanus' had quite different sequences, with similarity between 66.0 and 79.5%. These results confirm that the SPS-HLB liberibacter is a novel species for which the name 'Candidatus Liberibacter americanus' is proposed. Like the African and the Asian liberibacters, the 'American' liberibacter is restricted to the sieve tubes of the citrus host. The liberibacter could also be detected by PCR amplification of the 16S rRNA gene in Diaphorina citri, the psyllid vector of 'Ca. L. asiaticus', suggesting that this psyllid is also a vector of 'Ca. L. americanus' in SPS. 'Ca. L. americanus' was detected in 216 of 218 symptomatic leaf samples from 47 farms in 35 municipalities, while 'Ca. L. asiaticus' was detected in only 4 of the 218 samples, indicating that 'Ca. L. americanus' is the major cause of HLB in SPS. PMID:16166678

  12. Structural characterization of recombinant crustacyanin subunits from the lobster Homarus americanus.

    PubMed

    Ferrari, Michele; Folli, Claudia; Pincolini, Elisa; McClintock, Timothy S; Rössle, Manfred; Berni, Rodolfo; Cianci, Michele

    2012-08-01

    Crustacean crustacyanin proteins are linked to the production and modification of carapace colour, with direct implications for fitness and survival. Here, the structural and functional properties of the two recombinant crustacyanin subunits H(1) and H(2) from the American lobster Homarus americanus are reported. The two subunits are structurally highly similar to the corresponding natural apo crustacyanin CRTC and CRTA subunits from the European lobster H. gammarus. Reconstitution studies of the recombinant crustacyanin proteins H(1) and H(2) with astaxanthin reproduced the bathochromic shift of 85-95 nm typical of the natural crustacyanin subunits from H. gammarus in complex with astaxanthin. Moreover, correlations between the presence of crustacyanin genes in crustacean species and the resulting carapace colours with the spectral properties of the subunits in complex with astaxanthin confirmed this genotype-phenotype linkage. PMID:22869108

  13. Structural characterization of recombinant crustacyanin subunits from the lobster Homarus americanus

    PubMed Central

    Ferrari, Michele; Folli, Claudia; Pincolini, Elisa; McClintock, Timothy S.; Rössle, Manfred; Berni, Rodolfo; Cianci, Michele

    2012-01-01

    Crustacean crustacyanin proteins are linked to the production and modification of carapace colour, with direct implications for fitness and survival. Here, the structural and functional properties of the two recombinant crustacyanin subunits H1 and H2 from the American lobster Homarus americanus are reported. The two subunits are structurally highly similar to the corresponding natural apo crustacyanin CRTC and CRTA subunits from the European lobster H. gammarus. Reconstitution studies of the recombinant crustacyanin proteins H1 and H2 with astaxanthin reproduced the bathochromic shift of 85–95 nm typical of the natural crustacyanin subunits from H. gammarus in complex with astaxanthin. Moreover, correlations between the presence of crustacyanin genes in crustacean species and the resulting carapace colours with the spectral properties of the subunits in complex with astaxanthin confirmed this genotype–phenotype linkage. PMID:22869108

  14. Nutritional quality of the edible tissues of European lobster Homarus gammarus and American lobster Homarus americanus.

    PubMed

    Barrento, Sara; Marques, António; Teixeira, Bárbara; Vaz-Pires, Paulo; Nunes, Maria Leonor

    2009-05-13

    The clawed lobsters Homarus gammarus and Homarus americanus are high-priced and appreciated food items in southern Europe. From a nutritional point of view there is still limited information on the chemical composition of edible tissues (muscle, hepatopancreas, and gonads) of both species. Therefore, the aims of the present work were to study the proximate chemical composition, energy, fatty acid and amino acid profiles, and cholesterol content in each edible tissue of both species and to evaluate their nutritional quality. Differences were observed between tissues, sexes, and species. Muscle and gonads were rich in protein, whereas hepatopancreas had high fat, cholesterol, and energy contents. All edible tissues were valuable sources of essential amino acids. Contrary to the common belief that shellfish consumption should be reduced in a low-fat and -cholesterol diet, all edible tissues of clawed lobsters have a nutritious value perfectly compatible with nutritious and healthy diets. PMID:19334784

  15. [Polychaetes (Annelida: Polychaeta) epibiont on Spondylus americanus (Bivalvia: Spondylidae) from Mochima National Park, Venezuela].

    PubMed

    Liñero Arana, Ildefonso; Díaz Díaz, Oscar

    2006-09-01

    The polychaetes epibiontic on the mollusk Spondylus americanus Hermann, 1781 were extracted from mollusks hand-collected at a depth of 10-30 m in Mochima National Park, Venezuela (10 degrees 21'00" N-63 degrees 23'36" W), using scuba diving gear. Forty-three polychaete species were identified on the 32 bivalve specimens analyzed. The Serpulidae included 17 especies, Eunicidae six and Terebellidae four species. The most abundant species were Hydroides dirampha Mörch, 1863, Pileolaria militaris Claparède, 1868 (Serpulidae), and Notaulax nudicollis Krøyer, 1856 (Sabellidae). Their geographic affinitie were: 51.3% Atlantic, 28.2% widely distributed, 17.9% Amphiamericans, and 2.6% have a disjunct distribution.

  16. Sex steroid and prolactin profiles in male American black bears (Ursus americanus) during denning.

    PubMed

    Tsubota, T; Garshelis, D L; Nelson, R A; Bahr, J M

    1999-01-01

    Serum sex steroid and prolactin profiles were examined in the male American black bear, Ursus americanus during denning. Sera collected in December and the following March from 8 denning male black bears in Minnesota, U.S.A. were assayed for testosterone, estradiol-17 beta and prolactin. Eight bears were confirmed to be the denning mode based on a serum urea to creatinine ratio less than 10. Serum testosterone concentrations tended to increase from December to the subsequent March whereas serum estradiol-17 beta concentrations tended to decrease during this period. There were few changes in serum prolactin concentrations between December and March. These findings suggest that spermatogenesis and testicular steroidogenesis initiated during denning may be influenced by changes in serum sex steroid concentrations in the American black bear. PMID:10027172

  17. Fatal disseminated blastomycosis in a free-ranging American black bear (Ursus americanus).

    PubMed

    Dykstra, Jaclyn A; Rogers, Lynn L; Mansfield, Susan A; Wünschmann, Arno

    2012-11-01

    An aged, free-ranging, female, radio-collared American black bear (Ursus americanus) died after an approximately 5 month long period of weight loss. Gross necropsy findings included severe diffuse pyogranulomatous bronchopneumonia, marked granulomatous lymphadenitis of tracheobronchial lymph nodes and multiple intra-abdominal lymph nodes, chronic focal jejunal ulceration, and widespread alopecia. Histopathologic examination revealed abundant fungal organisms morphologically compatible with Blastomyces sp. within pyogranulomatous inflammatory lesions in the lungs, multiple lymph nodes, liver, kidneys, jejunum, and right adrenal gland. In addition, the haircoat had a mild infestation of chewing lice (Trichodectes pinguis euarctidos), and large numbers of rhabditid nematodes consistent with Pelodera sp. were histologically observed within hair follicles. PMID:23012379

  18. Sarcoptic mange and Pelodera dermatitis in an American black bear (Ursus americanus).

    PubMed

    Fitzgerald, Scott D; Cooley, Thomas M; Cosgrove, Melinda K

    2008-06-01

    An adult female free-ranging American black bear (Ursus americanus) was presented in poor body condition, with advanced skin disease. Skin changes included hair loss, lichenification, crusting, and focal erosions. Skin scrapings and histopathology identified two distinct parasitic conditions that were contributing to this animal's dermatitis. Large numbers of larvae, nymphs, and adults of Sarcoptes scabiei were present in the superficial epidermis, and nematodes consistent with Pelodera strongyloides were abundant within the hair follicles. This appears to be the first reported case of Pelodera dermatitis in a bear species, adding a new differential agent to the list for dermatitis in bears. The sarcoptic mange and poor body condition of this bear may have been contributing factors to the development of the opportunistic Pelodera infestation. PMID:18634219

  19. Laryngeal squamous cell carcinoma in a North American black bear (Ursus americanus).

    PubMed

    Rotstein, David S; Govett, Pamela; Wolfe, Barbara

    2005-09-01

    A 10-yr-old female black bear (Ursus americanus) at the North Carolina Zoological Park presented with a 2-wk history of intermittent coughing and stertor. The animal was anesthetized for examination, and at intubation, a large mass associated with the tonsils and larynx was observed obstructing the airway. Cardiopulmonary collapse occurred during the procedure, and efforts at resuscitation were ineffective. At necropsy, an 8 cm times 6 cm times 5 cm mass was present in the larynx, infiltrating the epiglottis and obstructing the airway. Regional lymph nodes were mildly enlarged and contained tan inspissated substance. Histologically, the neoplasm was composed of nests of keratinizing squamous epithelial cells with evidence of lymphatic invasion and metastases to regional lymph nodes. PMID:17312782

  20. A description of a new "Amoebozoan" isolated from the American lobster, Homarus americanus.

    PubMed

    Cole, Jeffrey; Anderson, O Roger; Tekle, Yonas I; Grant, Jessica; Kat, Laura A; Nerad, Thomas

    2010-01-01

    Our knowledge of the diversity of amoeboid protists is rapidly expanding as new and old habitats are more fully explored. In 2003, while investigating the cause of an amoeboid disease afflicting lobsters on the East Coast, samples were examined for the presence of amoebae from the carapace washings of the American lobster, Homarus americanus. During this survey a unique community of gymnamoebae was discovered. Among the new taxa discovered was a small Thecamoeba-like organism with a single posteriorly directed pseudopodium. Although resembling Parvamoeba rugata, this amoeba displayed distinctive morphology from that isolate or any other amoebozoan. Phylogenetic analysis shows this amoeba is distantly related to the Thecamoebidae. In this paper we describe the unique morphology of a second species of Parvamoeba and discuss its phylogenetic position with respect to the "Amoebozoa."

  1. Structural characterization of recombinant crustacyanin subunits from the lobster Homarus americanus.

    PubMed

    Ferrari, Michele; Folli, Claudia; Pincolini, Elisa; McClintock, Timothy S; Rössle, Manfred; Berni, Rodolfo; Cianci, Michele

    2012-08-01

    Crustacean crustacyanin proteins are linked to the production and modification of carapace colour, with direct implications for fitness and survival. Here, the structural and functional properties of the two recombinant crustacyanin subunits H(1) and H(2) from the American lobster Homarus americanus are reported. The two subunits are structurally highly similar to the corresponding natural apo crustacyanin CRTC and CRTA subunits from the European lobster H. gammarus. Reconstitution studies of the recombinant crustacyanin proteins H(1) and H(2) with astaxanthin reproduced the bathochromic shift of 85-95 nm typical of the natural crustacyanin subunits from H. gammarus in complex with astaxanthin. Moreover, correlations between the presence of crustacyanin genes in crustacean species and the resulting carapace colours with the spectral properties of the subunits in complex with astaxanthin confirmed this genotype-phenotype linkage.

  2. Structure of a two-CAP-domain protein from the human hookworm parasite Necator americanus

    SciTech Connect

    Asojo, Oluwatoyin A.

    2011-05-01

    The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite N. americanus refined to a resolution limit of 2.2 Å is presented. Major proteins secreted by the infective larval stage hookworms upon host entry include Ancylostoma secreted proteins (ASPs), which are characterized by one or two CAP (cysteine-rich secretory protein/antigen 5/pathogenesis related-1) domains. The CAP domain has been reported in diverse phylogenetically unrelated proteins, but has no confirmed function. The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite Necator americanus was refined to a resolution limit of 2.2 Å. The structure was solved by molecular replacement (MR) using Na-ASP-2, a one-CAP-domain ASP, as the search model. The correct MR solution could only be obtained by truncating the polyalanine model of Na-ASP-2 and removing several loops. The structure reveals two CAP domains linked by an extended loop. Overall, the carboxyl-terminal CAP domain is more similar to Na-ASP-2 than to the amino-terminal CAP domain. A large central cavity extends from the amino-terminal CAP domain to the carboxyl-terminal CAP domain, encompassing the putative CAP-binding cavity. The putative CAP-binding cavity is a characteristic cavity in the carboxyl-terminal CAP domain that contains a His and Glu pair. These residues are conserved in all single-CAP-domain proteins, but are absent in the amino-terminal CAP domain. The conserved His residues are oriented such that they appear to be capable of directly coordinating a zinc ion as observed for CAP proteins from reptile venoms. This first structure of a two-CAP-domain ASP can serve as a template for homology modeling of other two-CAP-domain proteins.

  3. Functional morphology of gill ventilation of the goosefish, Lophius americanus (Lophiiformes: Lophiidae).

    PubMed

    Farina, Stacy C; Bemis, William E

    2016-06-01

    The goosefish, Lophius americanus, is a dorso-ventrally compressed marine fish that spends most of its life sitting on the substrate waiting to ambush prey. Species in the genus Lophius have some of the slowest ventilatory cycles recorded in fishes, with a typical cycle lasting more than 90s. They have a large gill chamber, supported by long branchiostegal rays and ending in a siphon-like gill opening positioned underneath and behind the base of the pectoral fin. Our goals were to characterize the kinematics of gill ventilation in L. americanus relative to those of more typical ray-finned fishes, address previous assertions about ventilation in this genus, and describe the anatomy of the gill opening. We found that phase 1 of ventilation (during which both the buccal and gill chamber are expanding) is greatly increased in duration relative to that of typical ray-finned fishes (ranging from 62 to 127s), and during this phase, the branchiostegal rays are slowly expanding. This slow expansion is almost visually imperceptible, especially from a dorsal view. Despite this unusually long phase 1, the pattern of skeletal movements follows that of a typical actinopterygian, refuting previous assertions that Lophius does not use its jaws, suspensorium, and operculum during ventilation. When individuals were disturbed from the sediment, they tended to breathe more rapidly by decreasing the duration of phase 1 (to 18-30s). Dissections of the gill opening revealed a previously undocumented dorsal extension of the adductor hyohyoideus muscle, which passes from between the branchiostegal rays, through the ventro-medial wall of the gill opening, and to the dorsal midline of the body. This morphology of the adductor hyohyoideus shares similarities with that of many Tetraodontiformes, and we suggest that it may be a synapomorphy for Lophiiformes+Tetraodontiformes. The specialized anatomy and function of the gill chamber of Lophius represents extreme modifications that provide

  4. Development of a multiplex PCR assay to detect Edwardsiella tarda, Streptococcus parauberis, and Streptococcus iniae in olive flounder (Paralichthys olivaceus).

    PubMed

    Park, Seong Bin; Kwon, Kyoung; Cha, In Seok; Jang, Ho Bin; Nho, Seong Won; Fagutao, Fernand F; Kim, Young Kyu; Yu, Jong Earn; Jung, Tae Sung

    2014-01-01

    A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.

  5. A low molecular weight peptide from snow mold with epitopic homology to the winter flounder antifreeze protein.

    PubMed

    Newsted, W J; Polvi, S; Papish, B; Kendall, E; Saleem, M; Koch, M; Hussain, A; Cutler, A J; Georges, F

    1994-01-01

    Evidence for a small size protein (ca. 3500 kDa) exhibiting epitopic homology to the Atlantic winter flounder antifreeze protein (AFP) is found in the snow molds Coprinus psychromorbidus, Myriosclerotinia borealis, and Typhula incarnata. The protein shows strong cross-reactivity with antisera specific for the flounder AFP. Preliminary studies suggest that the protein is synthesized in response to lowering the culture temperature, and that it is membrane associated and, therefore, may function in an analogous capacity to the fish AFP. Also, the protein is shown to have antifreeze properties as determined by nuclear magnetic resonance microimaging experiments.

  6. Differences in energy expenditures and growth dilution explain higher PCB concentrations in male summer flounder

    USGS Publications Warehouse

    Madenjian, Charles P.; Jensen, Olaf P.; Rediske, Richard R.; O'Keefe, James P.; Vastano, Anthony R.; Pothoven, Steven A.

    2016-01-01

    Comparison of polychlorinated biphenyl (PCB) concentrations between the sexes of mature fish may reveal important behavioral and physiological differences between the sexes. We determined whole-fish PCB concentrations in 23 female summer flounder Paralichthys dentatus and 27 male summer flounder from New Jersey coastal waters. To investigate the potential for differences in diet or habitat utilization between the sexes, carbon and nitrogen stable isotope ratios were also determined. In 5 of the 23 female summer flounder, PCB concentrations in the somatic tissue and ovaries were determined. In addition, we used bioenergetics modeling to assess the contribution of the growth dilution effect to the observed difference in PCB concentrations between the sexes. Whole-fish PCB concentrations for females and males averaged 87 and 124 ng/g, respectively; thus males were 43% higher in PCB concentration compared with females. Carbon and nitrogen stable isotope ratios did not significantly differ between the sexes, suggesting that diet composition and habitat utilization did not vary between the sexes. Based on PCB determinations in the somatic tissue and ovaries, we predicted that PCB concentration of females would increase by 0.6%, on average, immediately after spawning due to release of eggs. Thus, the change in PCB concentration due to release of eggs did not explain the higher PCB concentrations observed in males. Bioenergetics modeling results indicated that the growth dilution effect could account for males being 19% higher in PCB concentration compared with females. Thus, the bulk of the observed difference in PCB concentrations between the sexes was not explained by growth dilution. We concluded that a higher rate of energy expenditure in males, stemming from greater activity and a greater resting metabolic rate, was most likely the primary driver for the observed difference in PCB concentrations between the sexes.

  7. Differences in Energy Expenditures and Growth Dilution Explain Higher PCB Concentrations in Male Summer Flounder.

    PubMed

    Madenjian, Charles P; Jensen, Olaf P; Rediske, Richard R; O'Keefe, James P; Vastano, Anthony R; Pothoven, Steven A

    2016-01-01

    Comparison of polychlorinated biphenyl (PCB) concentrations between the sexes of mature fish may reveal important behavioral and physiological differences between the sexes. We determined whole-fish PCB concentrations in 23 female summer flounder Paralichthys dentatus and 27 male summer flounder from New Jersey coastal waters. To investigate the potential for differences in diet or habitat utilization between the sexes, carbon and nitrogen stable isotope ratios were also determined. In 5 of the 23 female summer flounder, PCB concentrations in the somatic tissue and ovaries were determined. In addition, we used bioenergetics modeling to assess the contribution of the growth dilution effect to the observed difference in PCB concentrations between the sexes. Whole-fish PCB concentrations for females and males averaged 87 and 124 ng/g, respectively; thus males were 43% higher in PCB concentration compared with females. Carbon and nitrogen stable isotope ratios did not significantly differ between the sexes, suggesting that diet composition and habitat utilization did not vary between the sexes. Based on PCB determinations in the somatic tissue and ovaries, we predicted that PCB concentration of females would increase by 0.6%, on average, immediately after spawning due to release of eggs. Thus, the change in PCB concentration due to release of eggs did not explain the higher PCB concentrations observed in males. Bioenergetics modeling results indicated that the growth dilution effect could account for males being 19% higher in PCB concentration compared with females. Thus, the bulk of the observed difference in PCB concentrations between the sexes was not explained by growth dilution. We concluded that a higher rate of energy expenditure in males, stemming from greater activity and a greater resting metabolic rate, was most likely the primary driver for the observed difference in PCB concentrations between the sexes.

  8. Differences in Energy Expenditures and Growth Dilution Explain Higher PCB Concentrations in Male Summer Flounder

    PubMed Central

    Madenjian, Charles P.; Jensen, Olaf P.; Rediske, Richard R.; O’Keefe, James P.; Vastano, Anthony R.; Pothoven, Steven A.

    2016-01-01

    Comparison of polychlorinated biphenyl (PCB) concentrations between the sexes of mature fish may reveal important behavioral and physiological differences between the sexes. We determined whole-fish PCB concentrations in 23 female summer flounder Paralichthys dentatus and 27 male summer flounder from New Jersey coastal waters. To investigate the potential for differences in diet or habitat utilization between the sexes, carbon and nitrogen stable isotope ratios were also determined. In 5 of the 23 female summer flounder, PCB concentrations in the somatic tissue and ovaries were determined. In addition, we used bioenergetics modeling to assess the contribution of the growth dilution effect to the observed difference in PCB concentrations between the sexes. Whole-fish PCB concentrations for females and males averaged 87 and 124 ng/g, respectively; thus males were 43% higher in PCB concentration compared with females. Carbon and nitrogen stable isotope ratios did not significantly differ between the sexes, suggesting that diet composition and habitat utilization did not vary between the sexes. Based on PCB determinations in the somatic tissue and ovaries, we predicted that PCB concentration of females would increase by 0.6%, on average, immediately after spawning due to release of eggs. Thus, the change in PCB concentration due to release of eggs did not explain the higher PCB concentrations observed in males. Bioenergetics modeling results indicated that the growth dilution effect could account for males being 19% higher in PCB concentration compared with females. Thus, the bulk of the observed difference in PCB concentrations between the sexes was not explained by growth dilution. We concluded that a higher rate of energy expenditure in males, stemming from greater activity and a greater resting metabolic rate, was most likely the primary driver for the observed difference in PCB concentrations between the sexes. PMID:26794728

  9. Differences in Energy Expenditures and Growth Dilution Explain Higher PCB Concentrations in Male Summer Flounder.

    PubMed

    Madenjian, Charles P; Jensen, Olaf P; Rediske, Richard R; O'Keefe, James P; Vastano, Anthony R; Pothoven, Steven A

    2016-01-01

    Comparison of polychlorinated biphenyl (PCB) concentrations between the sexes of mature fish may reveal important behavioral and physiological differences between the sexes. We determined whole-fish PCB concentrations in 23 female summer flounder Paralichthys dentatus and 27 male summer flounder from New Jersey coastal waters. To investigate the potential for differences in diet or habitat utilization between the sexes, carbon and nitrogen stable isotope ratios were also determined. In 5 of the 23 female summer flounder, PCB concentrations in the somatic tissue and ovaries were determined. In addition, we used bioenergetics modeling to assess the contribution of the growth dilution effect to the observed difference in PCB concentrations between the sexes. Whole-fish PCB concentrations for females and males averaged 87 and 124 ng/g, respectively; thus males were 43% higher in PCB concentration compared with females. Carbon and nitrogen stable isotope ratios did not significantly differ between the sexes, suggesting that diet composition and habitat utilization did not vary between the sexes. Based on PCB determinations in the somatic tissue and ovaries, we predicted that PCB concentration of females would increase by 0.6%, on average, immediately after spawning due to release of eggs. Thus, the change in PCB concentration due to release of eggs did not explain the higher PCB concentrations observed in males. Bioenergetics modeling results indicated that the growth dilution effect could account for males being 19% higher in PCB concentration compared with females. Thus, the bulk of the observed difference in PCB concentrations between the sexes was not explained by growth dilution. We concluded that a higher rate of energy expenditure in males, stemming from greater activity and a greater resting metabolic rate, was most likely the primary driver for the observed difference in PCB concentrations between the sexes. PMID:26794728

  10. Effectiveness and immunomodulation of chemotherapeutants against scuticociliate Philasterides dicentrarchi in olive flounder.

    PubMed

    Harikrishnan, Ramasamy; Jin, Chang-Nam; Kim, Man-Chul; Kim, Ju-Sang; Balasundaram, Chellam; Heo, Moon-Soo

    2010-03-01

    Philasterides dicentrarchi is a histophagous scuticociliate causes fatal scuticociliatosis in farmed olive flounder Paralichthys olivaceus. The average monthly prevalence of scuticociliatosis due to P. dicentrarchi infections was increased from May to July (40+/-3.1% to 79.4+/-1.7%) and it decreased from August to November (63+/-2.3% to 30+/-2.6%) in olive flounder farms at Jeju Island, South Korea during 2000-2006. The prevalence of mixed infection along with Vibrio spp. bacterial infection was 49+/-7.2% than that of other mixed infection. At present no effective control measure for P. dicentrarchi infection has been described and large production losses continue. In the present study, formalin, hydrogen peroxide and Jenoclean chemotheraputants were used for bath treatment. Among Jenoclean at a low concentration of 50ppm proved effective. The results were confirmed with in vitro motility assessments and morphological changes scoring system in P. dicentrarchi. On the other hand, similar trend was noted following hydrogen peroxide treatment at this concentration, but formalin was only moderately effective. Either hydrogen peroxide or Jenoclean are the promising compounds effective at low concentrations with short application time for P. dicentrarchi. Therefore, these substances were evaluated on day 10, 20 and 30 for their ability to enhance innate immune response and disease resistance against P. dicentrarchi in olive flounder after chemotheraputants bath treatment with 100ppm for 30min per day. All the tested immune parameters were enhanced by treatment with Jenoclean, but not formalin and hydrogen peroxide. These findings suggest that Jenoclean bath treatment can be used for ensuring the heath of cultured marine fish against internal parasites such as P. dicentrarchi.

  11. Species profiles: Life histories and environmental requirements of coastal fishes and invertebrates (South Florida): Southern, gulf, and summer flounders. [Paralichthys lethostigma; Paralichthys albigutta; Paralichthys dentatus

    SciTech Connect

    Gilbert, C.R.

    1986-07-01

    The southern, gulf, and summer flounders live most of their lives in or near estuaries. Adults migrate into offshore waters during fall and winter, where spawning occurs from September to April. Eggs and newly-hatched larvae are carried inshore into estuaries, where further growth takes place. Age at sexual maturity for the three species has been estimated from 2 to 4 years, and maximum longevity from 3 to 10 years. Females apparently live slightly longer than males. Southern and summer flounders are estimated to live slightly longer than the smaller gulf flounder. Postlarvae eat mostly zooplankton, and larger individuals feed on fish and crustaceans. The summer and gulf flounders prefer a hard and/or sandy substrate, whereas the southern flounder is more common in a soft bottom of rich organic mud, clay, or silt. The southern flounder, unlike the other two species, is highly euryhaline, and frequently occurs in fresh water. Commercial catch statistics lump the three species together under the name ''flounder.'' Peak commercial catch in the Gulf of Mexico (in 1977) was 1.5 million lb, for a value of $561,000; peak catch in the south Atlantic region (also in 1977) was 11.4 million lb, for a value of $5,100,000.

  12. A transcriptomic analysis of American lobster (Homarus americanus) immune response during infection with the bumper car parasite Anophryoides haemophila.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Greenwood, Spencer J

    2013-06-01

    Anophryoides haemophila is an important protistan parasite of American lobster, Homarus americanus, as it has been found to infect lobsters in the wild as well as causing major losses of lobsters maintained in commercial holding facilities. Expression of over 14,500 H. americanus hepatopancreatic genes were monitored during an A. haemophila infection challenge in order to elucidate molecular mechanisms involved in the lobster immune response. One hundred and forty-five genes were found to be differentially expressed during infection. For many genes, this study is the first to link their expression to an immune response to a known lobster pathogen. Several of the genes have previously been linked to crustacean or invertebrate immune response including: several anti-lipopolysaccharide factor isoforms (ALFHa), acute phase serum amyloid protein A (SAA), a serine protease inhibitor, a toll-like receptor, several haemocyanin subunits, phagocyte signaling-impaired protein, vitelline membrane outer layer protein-1, trypsin, and a C-type lectin receptor. Microarray results were verified using RT-qPCR and agreement was good between the two methods. The expression of six ALFHa isoforms was monitored via microarray where ALFHa-1, ALFHa-2, ALFHa-4 and ALFHa-6 were differentially expressed while ALFHa-3 and ALFHa7 were not. RT-qPCR analysis confirmed that ALFHa-1, ALFHA-2 and ALFHa-4 expression increased during infection with a peak at 5-7weeks for ALFHa-1 and 10weeks for ALFHa-2 and ALFHa-4. This suggests that different ALFHa isoforms are temporally expressed during A. haemophila infection. Importantly, these results provide evidence that different ALFHa isoforms have more significant roles in responding to A. haemophila infection. Significant increases in SAA gene expression were also found, corroborating previous findings of increased SAA expression during Aerococcus viridans infections; highlighting the importance of SAA as a marker of H. americanus immune activation and

  13. Detection of lymphocystis disease virus in Japanese flounder Paralichthys olivaceus and other marine teleosts from northern China

    NASA Astrophysics Data System (ADS)

    Zhan, Wenbin; Li, Yongqin; Sheng, Xiuzhen; Xing, Jing; Tang, Xiaoqian

    2010-11-01

    We isolated a strain of lymphocystis disease virus (LCDV) from Japanese flounder ( Paralichthys olivaceus) cultured in northern China. Based on published sequences of major capsid protein (MCP) gene of LCDV-cn (GenBank: AF126405), we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization (ISH) to detect LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral supernatant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus (OFIV and RSIV), Iridovirus (CzIV and WIV), Ranavirus (TFV and FV3), and Lymphocystivirus (8 LCDV). Based on the alignment, we confirmed the PCR product was from Lymphocystivirus (GenBank accession number DQ279090 (LCDV-HD)). Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen, stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel’s black rockfish ( Sebastes schlegeli Higendorf), redwing sea robin ( Lepidotrigla microptera Günther) and turbot ( Scophthalmus maximus) using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method.

  14. Edwardsiella tarda Outer Membrane Protein C: An Immunogenic Protein Induces Highly Protective Effects in Flounder (Paralichthys olivaceus) against Edwardsiellosis.

    PubMed

    Liu, Fuguo; Tang, Xiaoqian; Sheng, Xiuzhen; Xing, Jing; Zhan, Wenbin

    2016-01-01

    Outer membrane protein C of Edwardsiella tarda is a major cell surface antigen and it was identified to be an immunogenic protein by Western blot using flounder (Paralichthys olivaceus) anti-recombinant OmpC (rOmpC), and anti-E. tarda antibodies. rOmpC tested the immune protective effect against E. tarda challenge in a flounder model and produced a relative percentage of survival rate of 85%. The immune response of flounder induced by rOmpC was investigated, and the results showed that: (1) the levels of specific serum antibodies induced by rOmpC were significantly higher than the control group after the second week after immunization, and the peak level occurred at week five after immunization; (2) rOmpC could induce the proliferation of sIg+ lymphocytes, and the peak levels of sIg+ lymphocytes in blood, spleen, and pronephros occurred at 4-5 weeks after immunization; and (3) the MHCIIα, CD4-1, IL-1β, IL-6 and TNF-α genes were significantly induced after being injected with rOmpC. Taken together, these results demonstrated that rOmpC could evoke highly protective effects against E. tarda challenge and induce strong innate immune response and humoral immune response of flounder, which indicated that OmpC was a promising vaccine candidate against E. tarda infection. PMID:27420049

  15. 50 CFR 648.107 - Conservation equivalent measures for the summer flounder party/charter and recreational fishery.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Conservation equivalent measures for the summer flounder party/charter and recreational fishery. 648.107 Section 648.107 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN...

  16. 77 FR 48916 - Fisheries of the Exclusive Economic Zone Off Alaska; Arrowtooth Flounder in the Bering Sea and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-15

    ... INFORMATION: NMFS manages the groundfish fishery in the (BSAI) exclusive economic zone according to the... National Oceanic and Atmospheric Administration 50 CFR Part 679 RIN 0648-XC129 Fisheries of the Exclusive Economic Zone Off Alaska; Arrowtooth Flounder in the Bering Sea and Aleutian Islands Management Area...

  17. Edwardsiella tarda Outer Membrane Protein C: An Immunogenic Protein Induces Highly Protective Effects in Flounder (Paralichthys olivaceus) against Edwardsiellosis

    PubMed Central

    Liu, Fuguo; Tang, Xiaoqian; Sheng, Xiuzhen; Xing, Jing; Zhan, Wenbin

    2016-01-01

    Outer membrane protein C of Edwardsiella tarda is a major cell surface antigen and it was identified to be an immunogenic protein by Western blot using flounder (Paralichthys olivaceus) anti-recombinant OmpC (rOmpC), and anti-E. tarda antibodies. rOmpC tested the immune protective effect against E. tarda challenge in a flounder model and produced a relative percentage of survival rate of 85%. The immune response of flounder induced by rOmpC was investigated, and the results showed that: (1) the levels of specific serum antibodies induced by rOmpC were significantly higher than the control group after the second week after immunization, and the peak level occurred at week five after immunization; (2) rOmpC could induce the proliferation of sIg+ lymphocytes, and the peak levels of sIg+ lymphocytes in blood, spleen, and pronephros occurred at 4–5 weeks after immunization; and (3) the MHCIIα, CD4-1, IL-1β, IL-6 and TNF-α genes were significantly induced after being injected with rOmpC. Taken together, these results demonstrated that rOmpC could evoke highly protective effects against E. tarda challenge and induce strong innate immune response and humoral immune response of flounder, which indicated that OmpC was a promising vaccine candidate against E. tarda infection. PMID:27420049

  18. 76 FR 53831 - Fisheries of the Northeastern United States; Summer Flounder, Scup, and Black Sea Bass Fisheries...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-30

    ... summer flounder, scup, and black sea bass specifications published on December 28, 2010 (75 FR 81498). An... for each coastal state from North Carolina to Maine. Following publication, an error was identified in... error. DATES: Effective August 30, 2011 through December 31, 2011. FOR FURTHER INFORMATION...

  19. Edwardsiella tarda and Aeromonas hydrophila isolated from diseased Southern flounder (Paralichthys lethostigma) are virulent to channel catfish and Nile tilapia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study is to identify bacterial pathogens isolated from diseased Southern flounder and determine their virulence to channel catfish and Nile tilapia. Twenty five Gram-negative bacteria isolates were recovered from five tissues (skin lesions, brain, liver, intestine, and posterior kidn...

  20. Flounder growth and production as indicators of the nursery value of marsh habitats in a Mediterranean lagoon

    NASA Astrophysics Data System (ADS)

    Franco, Anita; Fiorin, Riccardo; Zucchetta, Matteo; Torricelli, Patrizia; Franzoi, Piero

    2010-11-01

    Estuarine marshes are known as suitable nursery areas for many marine migrant fishes, such as flounder. The potential nursery value of such habitats was investigated in the Venice lagoon, by using growth and production of 0-group flounder as indicators. Size-frequency distribution analysis was performed on fish samples collected fortnightly, from March 2004 to June 2005, in two marsh sites, Dese and Tessera, differing in their origin and environmental conditions. Samples were mostly composed of juvenile individuals, belonging to 0- and 1-group cohorts (the latter being present in Tessera only). A higher total production, either annual or monthly, and faster growth of 0-group flounder was observed in Dese, associated to a higher ecological performance of 0-group individuals in this site, as indicated by the higher P:B ratio values. Dese is a site located in a marsh complex characterized by the relevant influence of a nearby river, and the observed higher potential nursery value of this marsh area with respect to the other is discussed in the light of the higher trophic status and other environmental conditions in this site. The production results were also compared to those from other estuarine environments commonly acknowledged as important nurseries for European flounder.

  1. EFFECTS OF LARVAL STOCKING DENSITY ON LABORATORY-SCALE AND COMMERICAL-SCALE PRODUCTION OF SUMMER FLOUNDER, PARALICHTHYS DENTATUS

    EPA Science Inventory

    Three experiments investigating larval stocking densities of summer flounder from hatch to metamorphosis, Paralichthys dentatus, were conducted at laboratory-scale (75-L aquaria) and at commercial scale (1,000-L tanks). Experiments 1 and 2 at commercial scale tested the densities...

  2. Anti-lipopolysaccharide factors in the American lobster Homarus americanus: Molecular characterization and transcriptional response to Vibrio fluvialis challenge

    PubMed Central

    Beale, K.M.; Towle, D.W.; Jayasundara, N.; Smith, C.M.; Shields, J.D.; Small, H.J.; Greenwood, S.J.

    2008-01-01

    Two partial mRNA sequences predicted to encode anti-lipopolysaccharide factors (ALFs) were identified among expressed sequence tags generated from the American lobster Homarus americanus and complete cDNA sequences were obtained from library clones. Comparison of the translated amino acid sequences to those publicly available confirmed similarity to arthropod anti-lipopolysaccharide factors. Both protein sequences, designated ALFHa-1 and ALFHa-2, contained an N-terminal signal peptide and two half-cysteines participating in a disulfide bridge, features conserved in other ALFs. Predicted secondary structures were similar to that described for the ALF from the horseshoe crab Limulus polyphemus. As part of an exploratory study of immunity in H. americanus, lobsters were injected with the bacterium Vibrio fluvialis and gill, hematopoietic, and hepatopancreas tissues were sampled for analysis of gene expression of ALFHa-1 and ALFHa-2 by quantitative PCR. The relative abundance of ALFHa-2 mRNA was not significantly affected by Vibrio injection in any of the three tissues tested. In contrast, ALFHa-1 mRNA levels in gills were increased by the treatment some 17-fold. Our results support a molecularly specific regulation of antimicrobial proteins in response to bacterial infection in H. americanus. PMID:19956341

  3. Epidemiological evidence for a differential effect of hookworm species, Ancylostoma duodenale or Necator americanus, on iron status of children.

    PubMed

    Albonico, M; Stoltzfus, R J; Savioli, L; Tielsch, J M; Chwaya, H M; Ercole, E; Cancrini, G

    1998-06-01

    Hookworms infect 1.3 billion people annually. Iron-deficiency anemia caused or exacerbated by intestinal blood loss is the major feature of hookworm infection, with such loss caused by the feeding of hookworms upon the intestinal mucosa. The hookworms Ancylostoma duodenale and Necator americanus cause major gastrointestinal blood loss. While relatively greater blood loss has been reported in clinical studies due to infection with A. duodenale, there has been no evidence that endemic A. duodenale infection has a greater impact than N. americanus infection upon the iron status of populations. The authors compared the degree of anemia and iron deficiency associated with these 2 hookworm species at the individual and community levels using a sample of 525 school children in grades 1-4 in Pemba Island, Tanzania, and controlling for infection intensities and other child characteristics through multiple regression. In the 492 children with hookworm-positive fecal cultures, hemoglobin and ferritin concentrations decreased with increasing proportions of A. duodenale. Among children with only N. americanus larvae, the prevalence of anemia was 60.5% and the prevalence of ferritin under 12 mcg/l was 33.1%. In children with 50% or higher proportions of A. duodenale larvae, the respective prevalences were 80.6% and 58.9%. Children from schools with high prevalences of A. duodenale infection had significantly worse iron deficiency and anemia than did children from low prevalence schools.

  4. First report of Taenia arctos (Cestoda: Taeniidae) from grizzly (Ursus arctos horribilis) and black bears (Ursus americanus) in North America.

    PubMed

    Catalano, Stefano; Lejeune, Manigandan; Verocai, Guilherme G; Duignan, Pádraig J

    2014-04-01

    The cestode Taenia arctos was found at necropsy in the small intestine of a grizzly (Ursus arctos horribilis) and a black bear (Ursus americanus) from Kananaskis Country in southwestern Alberta, Canada. The autolysis of the tapeworm specimens precluded detailed morphological characterization of the parasites but molecular analysis based on mitochondrial DNA cytochrome c oxidase subunit 1 gene confirmed their identity as T. arctos. This is the first report of T. arctos from definitive hosts in North America. Its detection in Canadian grizzly and black bears further supports the Holarctic distribution of this tapeworm species and its specificity for ursids as final hosts. Previously, T. arctos was unambiguously described at its adult stage in brown bears (Ursus arctos arctos) from Finland, and as larval stages in Eurasian elk (Alces alces) from Finland and moose (Alces americanus) from Alaska, USA. Given the morphological similarity between T. arctos and other Taenia species, the present study underlines the potential for misidentification of tapeworm taxa in previous parasitological reports from bears and moose across North America. The biogeographical history of both definitive and intermediate hosts in the Holarctic suggests an ancient interaction between U. arctos, Alces spp., and T. arctos, and a relatively recent host-switching event in U. americanus.

  5. First report of Taenia arctos (Cestoda: Taeniidae) from grizzly (Ursus arctos horribilis) and black bears (Ursus americanus) in North America.

    PubMed

    Catalano, Stefano; Lejeune, Manigandan; Verocai, Guilherme G; Duignan, Pádraig J

    2014-04-01

    The cestode Taenia arctos was found at necropsy in the small intestine of a grizzly (Ursus arctos horribilis) and a black bear (Ursus americanus) from Kananaskis Country in southwestern Alberta, Canada. The autolysis of the tapeworm specimens precluded detailed morphological characterization of the parasites but molecular analysis based on mitochondrial DNA cytochrome c oxidase subunit 1 gene confirmed their identity as T. arctos. This is the first report of T. arctos from definitive hosts in North America. Its detection in Canadian grizzly and black bears further supports the Holarctic distribution of this tapeworm species and its specificity for ursids as final hosts. Previously, T. arctos was unambiguously described at its adult stage in brown bears (Ursus arctos arctos) from Finland, and as larval stages in Eurasian elk (Alces alces) from Finland and moose (Alces americanus) from Alaska, USA. Given the morphological similarity between T. arctos and other Taenia species, the present study underlines the potential for misidentification of tapeworm taxa in previous parasitological reports from bears and moose across North America. The biogeographical history of both definitive and intermediate hosts in the Holarctic suggests an ancient interaction between U. arctos, Alces spp., and T. arctos, and a relatively recent host-switching event in U. americanus. PMID:24382413

  6. Distribution, condition, and growth of newly settled southern flounder ( Paralichthys lethostigma) in the Galveston Bay Estuary, TX

    NASA Astrophysics Data System (ADS)

    Glass, Lindsay A.; Rooker, Jay R.; Kraus, Richard T.; Holt, G. Joan

    2008-08-01

    Several flatfish species, including southern flounder ( Paralichthys lethostigma) recruit to estuaries during early life. Therefore, evaluation of estuarine sites and habitats that serve as nurseries is critical to conservation and management. The present study used density data in conjunction with biochemical condition and growth measurements to evaluate settlement sites used by southern flounder in the Galveston Bay Estuary (GBE). In 2005, beam-trawl collections were made in three major sections of the GBE (East Bay, Galveston Bay, West Bay). Three sites were sampled in each bay. Within each sampling site, replicate collections were taken from three habitats: 1) marsh edge (< 1 m depth), 2) intermediate zone (10-20 m from marsh interface; ˜ 1 m depth), and 3) bay zone (typically > 100 m from marsh interface; depth > 1 m). Average size of southern flounder collected was 12-19 mm standard length, and peak densities occurred in January and February. Catch data indicated that densities of southern flounder were significantly greater in East Bay (2.75 per 100 m 2) than in Galveston Bay (0.91 per 100 m 2) or in West Bay (0.45 per 100 m 2). Densities were statistically similar among habitats. Otolith-based estimates of age indicated that the majority of southern flounder collected were 35-45 days old and derived from early December to early January hatch-dates. Growth rates were similar among bays and among habitats, with the average growth rate being 0.40 mm day - 1 (range: 0.21-0.76 mm day - 1 ). RNA:DNA was above the established baseline value for nutritional stress, indicating that newly settled southern flounder in the GBE were in relatively high condition. Habitat-specific differences in RNA:DNA ratios were not observed; however, ratios were significantly lower in West Bay (average 8.0) than in East Bay (average 9.5) or in Galveston Bay (average 9.8), suggesting the condition of new recruits may vary spatially within the GBE. Findings from the current study

  7. Development of retinal structure and visual acuity in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Zhang, Xiu-Mei; Gao, Tian-Xiang; Arimoto, Takafumi

    2000-12-01

    The retinal structure and visual acuity in Japanese flounder Paralichthys olivaceus at different stages of development were examined by light microscopy. The resolving power of the retina, the visual axis and the best visual field were estimated based on the distribution of cone cells in the retina. The visual system of the larvae appears poorly developed at hatching. The larvae with total length (TL) of less than 10 mm, have single cones only and the eyes were well pigmented. At 10 11 mm TL, most single cones fused to form double cones, with the single and double cones forming a mosaic pattern. From larvae to early juvenile the retina stretches, the cones increase in diameter and rods increase in number. Based on the highest density of the cones in the ventro-temporal region, the visual axis was orientated upforward. The resolving power of the retina in 40 530 mm TL Japanese flounder was found to range from 25.1 to 11.5 min. The results indicated continual improvements in the visual system of the growing fish towards higher resolving power, visual acuity and sensitivity.

  8. Expression of let-7 microRNAs that are involved in Japanese flounder (Paralichthys olivaceus) metamorphosis.

    PubMed

    Fu, Yuanshuai; Shi, Zhiyi; Wang, Guyue; Zhang, Junling; Li, Wenjuan; Jia, Liang

    2013-06-01

    The let-7 microRNAs (miRNAs), a class of small noncoding RNAs, are phylogenetically conserved and temporally expressed and control the proper timing of events during development as heterochronic genes in many animals. Japanese flounder (Paralichthys olivaceus) undergoes a metamorphosis from the larval to juvenile form. Here, we identified 21 let-7 miRNA precursors from different genome loci in Japanese flounder. P. olivaceus let-7 miRNAs are widely expressed in adult tissues, highly expressed during metamorphosis, but weakly during embryonic development. Exogenous thyroid hormone (0.1 mg/L), which induces premature metamorphosis, significantly promotes the expression of let-7 miRNAs, while thiourea (30 mg/L), which affects metamorphic arrest, inhibits the expression of let-7 miRNAs in metamorphosis in P. olivaceus. These results show that let-7 miRNAs widely participate in tissue development and metabolism during development and are also involved in regulation of temporal transitions associated with cell proliferation and differentiation during metamorphosis, in P. olivaceus.

  9. The renin-angiotensin system and drinking in the euryhaline flounder, Platichthys flesus.

    PubMed

    Carrick, S; Balment, R J

    1983-09-01

    Drinking behaviour and its possible regulation by the renin-angiotensin system (RAS) has been examined in the euryhaline flounder. Fluid intake was greater in seawater (SW)-adapted than freshwater (FW)-adapted fish, the latter having significantly lower plasma sodium, chloride, and osmotic concentrations. Oesophageal cannulation in SW-adapted fish resulted in further elevation of drinking rates, which increased proportionally with progressive body water loss as measured by the fall in body weight and rise in plasma tonicity. The influence of the RAS on drinking in SW-adapted fish was examined in animals with an intact gastrointestinal tract. Fluid intake fell markedly following administration of the converting enzyme inhibitor, Captopril. Infusions of angiotensin I (AI) and angiotensin II (AII) induced dose-related increments in the rate of drinking. The increased drinking in response to AI was inhibited, however, by the simultaneous administration of Captopril. The results are consistent with the presence in the flounder of the major elements of the RAS, including AI, AII, and a converting enzyme-like substance. The RAS appears to play an important regulatory role in the adaptative drinking behaviour associated with migration of euryhaline teleosts between FW and SW.

  10. Transcriptomic responses of European flounder (Platichthys flesus) liver to a brominated flame retardant mixture.

    PubMed

    Williams, Tim D; Diab, Amer M; Gubbins, Matt; Collins, Catherine; Matejusova, Iveta; Kerr, Rose; Chipman, James K; Kuiper, Raoul; Vethaak, A Dick; George, Stephen G

    2013-10-15

    Male European flounder (Platichthys flesus) were exposed to a technical mixture of brominated diphenyl ethers (PDBEs, DE-71, Pentamix) that had been purified to remove contaminating dioxins. Controls were exposed to carrier solvent alone. Fish were exposed to decadally increasing concentrations of Pentamix via both sediment and spiked food. The GENIPOL P. flesus cDNA microarray, differentially expressed gene profiling (DEG) and quantitative PCR were employed to detect hepatic transcriptional differences between exposed fish and controls. Gene transcriptional changes were more sensitive to Pentamix exposure than biomarkers measured previously. Pentamix exposure induced transcripts coding for enzymes of xenobiotic metabolism (CYP1A, aldo-keto reductases) and elicited endocrine disruption (vitellogenin and thyroid hormone receptor alpha), with effects on CYP1A and VTG occurring at the highest exposure. Ontology analysis clearly showed dose-responsive changes indicative of oxidative stress, induction of mitochondrial dysfunction, and apoptosis. We conclude that exposure to PBDEs in both sediment and food has a significant adverse effect on a broad range of crucial biochemical processes in the livers of this widely distributed estuarine fish species, the flounder. PMID:23948077

  11. Spatial and temporal variability in growth of southern flounder (Paralichthys lethostigma)

    USGS Publications Warehouse

    Midway, Stephen R.; Wagner, Tyler; Arnott, Stephen A.; Biondo, Patrick; Martinez-Andrade, Fernando; Wadsworth, Thomas F.

    2015-01-01

    Delineation of stock structure is important for understanding the ecology and management of many fish populations, particularly those with wide-ranging distributions and high levels of harvest. Southern flounder (Paralichthys lethostigma) is a popular commercial and recreational species along the southeast Atlantic coast and Gulf of Mexico, USA. Recent studies have provided genetic and otolith morphology evidence that the Gulf of Mexico and Atlantic Ocean stocks differ. Using age and growth data from four states (Texas, Alabama, South Carolina, and North Carolina) we expanded upon the traditional von Bertalanffy model in order to compare growth rates of putative geographic stocks of southern flounder. We improved the model fitting process by adding a hierarchical Bayesian framework to allow each parameter to vary spatially or temporally as a random effect, as well as log transforming the three model parameters (L∞, K, andt0). Multiple comparisons of parameters showed that growth rates varied (even within states) for females, but less for males. Growth rates were also consistent through time, when long-term data were available. Since within-basin populations are thought to be genetically well-mixed, our results suggest that consistent small-scale environmental conditions (i.e., within estuaries) likely drive growth rates and should be considered when developing broader scale management plans.

  12. Possible Involvement of Photoperiodic Regulation in Reproductive Endocrine System of Female Olive Flounder Paralichthys olivaceus.

    PubMed

    Kim, Hyun Chul; Lee, Chi Hoon; Hur, Sung Pyu; Kim, Byeong Hoon; Park, Jun Young; Lee, Young Don

    2015-03-01

    This study investigated possible involvement of photoperiodic regulation in reproductive endocrine system of female olive flounder. To investigate the influence on brain-pituitary axis in endocrine system by regulating photoperiod, compared expression level of Kisspeptin and sbGnRH mRNA in brain and FSH-β, LH-β and GH mRNA in pituitary before and after spawning. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from Aug. 2013 to Jun. 2014. Continuous long photoperiod treatment from Aug. (post-spawning phase) was inhibited gonadal development of female olive flounder. In natural photoperiod group, the Kiss2 expression level a significant declined in Mar. (spawning period). And also, FSH-β, LH-β and GH mRNA expression levels were increasing at this period. However, in long photoperiod group, hypothalamic Kiss2, FSH-β, LH-β and GH mRNA expression levels did not show any significant fluctuation. These results suggest that expression of hypothalamic Kiss2, GtH and GH in the pituitary would change in response to photoperiod and their possible involvement of photoperiodic regulation in reproductive endocrine system of the BPG axis.

  13. A Vibrio anguillarum strain associated with skin ulcer on cultured flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Mo, Zhao-Lan; Tan, Xun-Gang; Xu, Yong-Li; Zhang, Pei-Jun

    2001-12-01

    The characteristics of a bacterium strain M3, isolated from cultured flounder Paralichthys olivaceus with remarkable external sign of skin ulcer during an epizootic outbreak, indicated that the bacterium belonged to the species Vibrio anguillarum. Challenge by I.M. (intramuscular injection), bath, and oral administration with M3 showed that it was highly pathogenic for Paralichthys olivacues. The LD50 dose was 5.144×103 CFU/ per fish infection by I.M. injection. Recovered inoculated bacteria from the surviving fish revealed that the asymptomatic carriers could be a latent contagious source. Study of the effect of bacterial culture CFS (cell-free-supernatant) showed that the exotoxins produced by M3 play an important role in its pathogenicity for flounder. The resistance of M3 to 36 out of 41 antibiotics indicated that the bacterial disease outbreak was mainly attributable to the frequent and excessive use of antimicrobial agents; and that vaccination would be an effective precaution against bacterial disease.

  14. Diel feeding patterns and daily food intake of juvenile stone flounder Platichthys bicoloratus

    NASA Astrophysics Data System (ADS)

    Tomiyama, Takeshi; Katayama, Satoshi; Yamamoto, Masayuki; Shoji, Jun

    2016-01-01

    Pleuronectid flatfish are considered visual day feeders, but their ability to feed at night has not been examined in the field. Juvenile (age-0) stone flounder Platichthys bicoloratus were collected, and their stomach contents were investigated in situ every 4 h over a 24-h period in an estuarine habitat to elucidate diel feeding periodicity. The weight of juvenile stomach contents was usually the highest around dusk and the main prey was bivalve siphons. To reveal whether juveniles feed only at particular times during the day or throughout a 24-h period, we conducted 24-h cage experiments in which 10 juveniles with empty stomachs were held in a cage for approximately 4 h (six trials). This experiment was carried out three times during different moon phases. Juveniles primarily ingested prey during the day, but 30% of fish that were caged at night also ingested prey. The number of successful captures by the caged fish was much greater during the day than that at night. These results indicate that stone flounder generally feed during the day and they may only feed at night under unusual situations, although they have the ability to capture prey at night. The mean daily ration estimated by diel changes in stomach content weight varied from 3.4% (95% confidence interval, 1.8-4.8%) to 13.2% (11.0-15.6%) of body weight between survey dates, indicating that daily food consumption by fish estimated from a single survey may be strongly biased.

  15. Kinetics and thermodynamics of lactate dehydrogenases from beef heart, beef muscle, and flounder muscle.

    PubMed

    Borgmann, U; Laidler, K J; Moon, T W

    1975-11-01

    The eight rate constants for a four-step ordered ternary-complex mechanism have been compared for lactate dehydrogenases (EC1.1.1.27) from three sources, beef heart, beef muscle, and flounder muscle. The rate constants were determined at temperatures ranging from 5 degrees C to 50 degrees C, and the corresponding activation parameters deltaG not equal to, deltaH not equal to, and deltaS not equal to were calculated. Significant differences are noted for the values for the three types of enzyme. The relative heights of the activation barriers are much the same in all three cases, differences in kinetic behavior resulting mainly from differences in the stable binary and ternary enzyme-substrate complexes. These complexes are, in general, at lower free-energy and enthalpy levels of the beef-heart and beef-muscle enzymes than for the flounder-muscle enzyme. A high degree of compensation is found between the enthalpies and entropies of activation, resulting in relatively small differences between the free energies (and rates) for homologous steps with different enzymes. Analysis of the results, on the assumption that the compensation effect is due to weak-bonding effects, suggests that there are fewer weak bonds in the stable complexes of the muscle enzymes.

  16. Ion and water transport by the flounder urinary bladder: salinity dependence.

    PubMed

    Demarest, J R

    1984-04-01

    Urinary bladders from seawater-acclimated (SW) flounder had a transepithelial resistance (Rt) of congruent to 2,000 omega X cm2 and absorbed Na and Cl at equal rates of about 3 mueq X cm-2 X h-1 in an electrically silent manner; the short-circuit current (Isc) was 0.03 +/- 0.01 mueq X cm-2 X h-1. The transport of Na and Cl was only partially coupled. Removal of Na (or Cl) from the bathing solutions reduced net Cl (or Na) absorption by only 60%, yet there was neither a change in transepithelial potential nor the appearance of a short-circuit current that could be associated with the net Cl (or Na) transport that remained. Bladders from freshwater-acclimated (FW) flounder had a fivefold lower Rt and exhibited the same partially coupled and equal Na and Cl transport, but the ion transport rates were twice as large as those of SW bladders and the bladders exhibited a significant Isc of 0.51 +/- 0.08 mueq X cm-2 X h-1. The rate of fluid transport was much lower in FW than in SW bladders, reflecting a sixfold decrease in hydraulic conductivity (Lp). In both SW and FW bladders a large portion of the serosal-to-mucosal ion movement appears to be through nonconductive pathways.

  17. Possible Involvement of Photoperiodic Regulation in Reproductive Endocrine System of Female Olive Flounder Paralichthys olivaceus

    PubMed Central

    Kim, Hyun Chul; Lee, Chi Hoon; Hur, Sung Pyu; Kim, Byeong Hoon; Park, Jun Young; Lee, Young Don

    2015-01-01

    This study investigated possible involvement of photoperiodic regulation in reproductive endocrine system of female olive flounder. To investigate the influence on brain-pituitary axis in endocrine system by regulating photoperiod, compared expression level of Kisspeptin and sbGnRH mRNA in brain and FSH-β, LH-β and GH mRNA in pituitary before and after spawning. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from Aug. 2013 to Jun. 2014. Continuous long photoperiod treatment from Aug. (post-spawning phase) was inhibited gonadal development of female olive flounder. In natural photoperiod group, the Kiss2 expression level a significant declined in Mar. (spawning period). And also, FSH-β, LH-β and GH mRNA expression levels were increasing at this period. However, in long photoperiod group, hypothalamic Kiss2, FSH-β, LH-β and GH mRNA expression levels did not show any significant fluctuation. These results suggest that expression of hypothalamic Kiss2, GtH and GH in the pituitary would change in response to photoperiod and their possible involvement of photoperiodic regulation in reproductive endocrine system of the BPG axis. PMID:25949205

  18. Analysis of gene expression in Homarus americanus larvae exposed to sublethal concentrations of endosulfan during metamorphosis.

    PubMed

    Bauer, Megan; Greenwood, Spencer J; Clark, K Fraser; Jackman, Paula; Fairchild, Wayne

    2013-12-01

    Agricultural pesticide runoff has been suspected as the cause of numerous fish kills in rivers throughout Prince Edward Island but the impact on the surrounding marine environment is unknown. Endosulfan, an organochlorine pesticide, is a potent neurotoxin and molt inhibitor used to combat the Colorado potato beetle however it has the potential to affect non-target organisms including the American lobster (Homarus americanus). Metamorphosis is a critical stage of development and the effects of contaminant exposure during this time are largely unknown in lobster. A 14day endosulfan exposure was performed to identify the effects on survival, development and gene expression in lobster larvae during metamorphosis; all of which were predicted to be negatively impacted. The higher endosulfan concentrations resulted in greater mortality and a significant increase in the number of days required to reach metamorphosis in surviving animals. A custom made H. americanus microarray was used for monitoring the changes in expression of 14,592 genes at the termination of the exposure. Genes with >1.5 fold change and identified as being significant at p<0.05 using one-way ANOVA were selected for further analysis. A total of 707 genes were identified as being significantly differentiated, however with only ~40% annotation of the array, the majority of these genes were unknown. Annotated genes of interest were involved in many processes: development, metabolism, immunity and oxidative stress response and gene regulation. Nine genes of interest (histone H1, farnesoic acid O-methyltransferase, cuticle protein, glutathione S-transferase, thioredoxin, NADH dehydrogenase, ecdysone nuclear receptor Fushi tarazu F1 (FTZ-F1), ferritin and ecdysone inducible protein E75 (EIP-E75)) were selected for RT-qPCR validation of the microarray results. The RT-qPCR method was more sensitive than the microarray yet detected similar expression patterns. The two highest endosulfan concentrations resulted

  19. Analysis of gene expression in Homarus americanus larvae exposed to sublethal concentrations of endosulfan during metamorphosis.

    PubMed

    Bauer, Megan; Greenwood, Spencer J; Clark, K Fraser; Jackman, Paula; Fairchild, Wayne

    2013-12-01

    Agricultural pesticide runoff has been suspected as the cause of numerous fish kills in rivers throughout Prince Edward Island but the impact on the surrounding marine environment is unknown. Endosulfan, an organochlorine pesticide, is a potent neurotoxin and molt inhibitor used to combat the Colorado potato beetle however it has the potential to affect non-target organisms including the American lobster (Homarus americanus). Metamorphosis is a critical stage of development and the effects of contaminant exposure during this time are largely unknown in lobster. A 14day endosulfan exposure was performed to identify the effects on survival, development and gene expression in lobster larvae during metamorphosis; all of which were predicted to be negatively impacted. The higher endosulfan concentrations resulted in greater mortality and a significant increase in the number of days required to reach metamorphosis in surviving animals. A custom made H. americanus microarray was used for monitoring the changes in expression of 14,592 genes at the termination of the exposure. Genes with >1.5 fold change and identified as being significant at p<0.05 using one-way ANOVA were selected for further analysis. A total of 707 genes were identified as being significantly differentiated, however with only ~40% annotation of the array, the majority of these genes were unknown. Annotated genes of interest were involved in many processes: development, metabolism, immunity and oxidative stress response and gene regulation. Nine genes of interest (histone H1, farnesoic acid O-methyltransferase, cuticle protein, glutathione S-transferase, thioredoxin, NADH dehydrogenase, ecdysone nuclear receptor Fushi tarazu F1 (FTZ-F1), ferritin and ecdysone inducible protein E75 (EIP-E75)) were selected for RT-qPCR validation of the microarray results. The RT-qPCR method was more sensitive than the microarray yet detected similar expression patterns. The two highest endosulfan concentrations resulted

  20. Cytochrome P4501A indices as biomarkers of contaminant exposure: A field study with plaice and flounder

    SciTech Connect

    Eggens, M.; Opperhuizen, A.

    1995-12-31

    Samples of two flatfish species, flounder (Platichthys flesus) and plaice (Pleurnectus platessa), were collected during August/September 1991 and May/June 1992 in the southern North Sea. The induction of hepatic cytochrome P4501A1 (CYP1A) in both species was examined, when exposed to environmental PCBs and PAHs. CYP1A was measured by a semi-quantitative ELISA technique and by the activity of its catalyst, 7-ethoxyresorufin O-deethylase (EROD). In plaice, both hepatic CYP1 protein level and EROD activity were significantly higher at offshore sampling locations than at coastal locations, by a factor of 1.5 to 2. In flounder, which were only collected at coastal locations, the range of values of EROD activity was much greater than in plaice, but the range in CYP1A protein content was comparable in both species. multiple regression analysis of data from plaice and flounder did not show any significant correlation between CYP1A indices, water temperature and contaminant concentrations in fish tissues. CYP1A indices also appeared to be unrelated to food type, as determined by visual screening of fish intestines. It is concluded that the induction of CYP1A in plaice and flounder from the southern North Sea is not related in a simple manner to exposure to PCBs and PAHs. From the point of view-of use as a biomarker, the measurement of CYP1A protein level and EROD activity in plaice and flounder from the open North Sea cannot be applied straightforwardly for monitoring exposure to PCBs and/or PAHs.

  1. Fecundity regulation in relation to habitat utilisation of two sympatric flounder (Platichtys flesus) populations in the brackish water Baltic Sea

    NASA Astrophysics Data System (ADS)

    Nissling, Anders; Thorsen, Anders; da Silva, Filipa F. G.

    2015-01-01

    Two populations of flounder (Platichtys flesus) with different life history traits inhabit the brackish water Baltic Sea. Both types share feeding areas in coastal waters during summer-autumn but utilise different habitats for spawning in spring, namely offshore spawning with pelagic eggs and coastal spawning with demersal eggs respectively. Fecundity regulation by atresia was assessed as prevalence (portion of fish with atresia) and intensity (calculated as the average intensity of atresia in these fish) during the reproductive cycle following start of gonad development in the autumn up to spawning in spring, and evaluated in relation to fish condition (Fulton's condition factor reflecting energy reserves of the fish) and feeding incidence of the respective population. Peaking in winter (December-February), fecundity regulation was significantly higher for coastal spawning flounder than for flounder spawning offshore. For coastal spawners, the prevalence was 45-90% with an intensity of 6.4-9.3% vs. 0-25% and an intensity of 2.1-3.4% for offshore spawners during winter. Further, fecundity regulation ceased prior to spawning for offshore spawners but continued for coastal spawners. For coastal spawners, the prevalence was 12-29% and an intensity of 2.5-6.1% during spawning. The change in fish condition was strongly related to feeding incidence and differed between populations. As feeding ceased, condition of offshore spawners decreased during winter up to spawning, whereas condition of coastal spawners decreased during autumn but was maintained as feeding started again prior to spawning. Thus, habitat utilisation according to spawning strategy affects the timing of fecundity down-regulation reflecting availability of resources, namely limited food resources in deep areas and higher availability in coastal areas. Offshore spawning flounder display characteristics typical for a capital spawner with ceasing of feeding and oocyte down-regulation well before spawning

  2. Immune responses of flounder Paralichthys olivaceus vaccinated by immersion of formalin-inactivated Edwardsiella tarda following hyperosmotic treatment.

    PubMed

    Gao, Ying-Li; Tang, Xiao-Qian; Sheng, Xiu-Zhen; Xing, Jing; Zhan, Wen-Bin

    2015-10-16

    The aim of the present study was to evaluate the effects of hyperosmotic immersion (HI) vaccination and determine the optimum hyperosmotic salinity for flounder Paralichthys olivaceus by investigating its immune responses following vaccination. Flounder were immersed in 1 of 3 hyperosmotic solutions at 50, 60 and 70‰ salinity, then transferred into 30‰ salinity normal seawater containing formalin-inactivated Edwardsiella tarda for vaccination (3 HI groups), or were immersed in normal seawater as direct immersion (DI group). The results showed that the percentages of surface membrane immunoglobulin-positive (sIg+) cells in peripheral blood leukocytes and spleen leukocytes induced by HI were significantly higher than that with DI (p < 0.05), and the 50‰ salinity group showed the strongest response among the HI groups, which reached peaks at Week 4. ELISA assay showed that the specific serum antibodies gradually increased after vaccination and reached peak at Day 32, and the fish treated with HI showed stronger antibody responses; among the HI groups, a significantly higher specific antibody level was detected in the 50‰ salinity group at Day 32 (p < 0.05). Similarly, the fish treated with HI showed higher specific mucosal antibody levels compared to the DI group, and the mucosal antibody showed a faster response, with peak time arriving 1 wk earlier than for the serum antibody. The relative percent survival (RPS) of flounder treated with HI at 50, 60 and 70‰ salinities were 79, 71 and 57% respectively, while this was 43% in the DI group. These results demonstrated that HI, especially the 50‰ salinity, could efficiently enhance the immune response of flounder and show higher RPS. This has significant value for immunological prevention of edwardsiellosis in flounder. PMID:26480914

  3. Meteorological effects on the biting activity of Leptoconops americanus(Diptera: Ceratopogonidae).

    PubMed

    Strickman, D; Wirtz, R; Lawyer, P; Glick, J; Stockwell, S; Perich, M

    1995-03-01

    Collections of biting Leptoconops americanus were made at half-hour intervals throughout the daylight hours on Stansbury Island, UT, during 9 days in May, 1993. The most favorable conditions for biting (> or = 90 bites on the ears in 15 min) included temperatures higher than 15 degrees C, minimum wind (< 5 kph), minimum cloud cover, maximum sun, and no rain. Temperatures below 10 degrees C or the presence of rain prevented almost all biting. Higher winds and cloudiness decreased biting activity, but did not eliminate it if other conditions were favorable. Although not statistically significant, there was some suggestion from the data that higher temperatures (> 25 degrees C) reduced biting. The flies did not appear to be more numerous at any particular part of the day; the biting rate simply followed meteorological conditions at the time. Ambient light varied between 1 and 10,000 foot candles during the study, with high biting rates (76 and 99 bites per 15 min) observed at levels as low as 80-100 foot candles.

  4. Serosurvey for selected pathogens in free-ranging American black bears (Ursus americanus) in Maryland, USA.

    PubMed

    Bronson, Ellen; Spiker, Harry; Driscoll, Cindy P

    2014-10-01

    American black bears (Ursus americanus) in Maryland, USA, live in forested areas in close proximity to humans and their domestic pets. From 1999 to 2011, we collected 84 serum samples from 63 black bears (18 males; 45 females) in five Maryland counties and tested them for exposure to infectious, including zoonotic, pathogens. A large portion of the bears had antibody to canine distemper virus and Toxoplasma gondii, many at high titers. Prevalences of antibodies to zoonotic agents such as rabies virus and to infectious agents of carnivores including canine adenovirus and canine parvovirus were lower. Bears also had antibodies to vector-borne pathogens common to bears and humans such as West Nile virus, Borrelia burgdorferi, Rickettsia rickettsii, and Anaplasma phagocytophilum. Antibodies were detected to Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Canicola, Grippotyphosa, and Bratislava. We did not detect antibodies to Brucella canis or Ehrlichia canis. Although this population of Maryland black bears demonstrated exposure to multiple pathogens of concern for humans and domesticated animals, the low levels of clinical disease in this and other free-ranging black bear populations indicate the black bear is likely a spillover host for the majority of pathogens studied. Nevertheless, bear populations living at the human-domestic-wildlife interface with increasing human and domestic animal exposure should continue to be monitored because this population likely serves as a useful sentinel of ecosystem health.

  5. Seasonal variation in American black bear Ursus americanus activity patterns: Quantification via remote photography

    USGS Publications Warehouse

    Bridges, A.S.; Vaughan, M.R.; Klenzendorf, S.

    2004-01-01

    Activity pattern plasticity may serve as an evolutionary adaptation to optimize fitness in an inconstant environment, however, quantifying patterns and demonstrating variation can be problematic. For American black bears Ursus americanus, wariness and habitat inaccessibility further complicate quantification. Radio telemetry has been the primary technique used to examine activity, however, interpretation error and limitation on numbers of animals available to monitor prevent extrapolation to unmarked or untransmittered members of the population. We used remote cameras to quantify black bear activity patterns and examined differences by season, sex and reproductive class in the Alleghany Mountains of western Virginia, USA. We used 1,533 pictures of black bears taken during 1998-2002 for our analyses. Black bears generally were diurnal in summer and nocturnal in autumn with a vespertine activity peak during both seasons. Bear-hound training seasons occurred during September and may offer explanation for the observed shift towards nocturnal behaviour. We found no substantial differences in activity patterns between sex and reproductive classes. Use of remote cameras allowed us to efficiently sample larger numbers of individual animals and likely offered a better approximation of population-level activity patterns than individual-level, telemetry-based methodologies.

  6. Social network analysis of mating patterns in American black bears (Ursus americanus).

    PubMed

    Moore, Jennifer A; Xu, Ran; Frank, Kenneth; Draheim, Hope; Scribner, Kim T

    2015-08-01

    Nonrandom mating can structure populations and has important implications for population-level processes. Investigating how and why mating deviates from random is important for understanding evolutionary processes as well as informing conservation and management. Prior to the implementation of parentage analyses, understanding mating patterns in solitary, elusive species like bears was virtually impossible. Here, we capitalize on a long-term genetic data set collected from black bears (Ursus americanus) (N = 2422) in the Northern Lower Peninsula (NLP) of Michigan, USA. We identified mated pairs using parentage analysis and applied logistic regression (selection) models that controlled for features of the social network, to quantify the effects of individual characteristics, and spatial and population demographic factors on mating dynamics. Logistic regression models revealed that black bear mating was associated with spatial proximity of mates, male age, the time a pair had coexisted, local population density and relatedness. Mated pairs were more likely to contain older males. On average, bears tended to mate with nearby individuals to whom they were related, which does not support the existence of kin recognition in black bears. Pairwise relatedness was especially high for mated pairs containing young males. Restricted dispersal and high male turnover from intensive harvest mortality of NLP black bears are probably the underlying factors associated with younger male bears mating more often with female relatives. Our findings illustrate how harvest has the potential to disrupt the social structure of game species, which warrants further attention for conservation and management. PMID:26113220

  7. Aroma components of cooked tail meat of American lobster (Homarus americanus).

    PubMed

    Lee, G H; Suriyaphan, O; Cadwallader, K R

    2001-09-01

    Key aroma components of cooked tail meat of American lobster (Homarus americanus) were studied by gas chromatography-olfactometry (GCO) techniques. Components of low and intermediate volatility were evaluated by aroma extract dilution analysis of solvent extracts prepared by direct solvent extraction-high vacuum distillation and vacuum steam distillation-solvent extraction, whereas headspace volatile components were assessed by GCO of decreasing headspace (static and dynamic modes) samples. Forty-seven odorants were detected by all techniques. 3-Methylbutanal (chocolate, malty), 2,3-butanedione (buttery), 3-(methylthio)propanal (cooked potato), 1-octen-3-one (mushroom), 2-acetyl-1-pyrroline (popcorn), and (E,Z)-2,6-nonadienal (cucumber), were identified as predominant odorants by all four isolation methods. The highly volatile compounds methanethiol (rotten, sulfurous) and dimethyl sulfide (canned corn) were detected by headspace methods only. These eight odorants along with three unknown compounds with crabby, amine, fishy odors were found to predominate in the overall aroma of cooked lobster tail meat. PMID:11559132

  8. Bone formation is not impaired by hibernation (disuse) in black bears Ursus americanus

    USGS Publications Warehouse

    Donahue, S.W.; Vaughan, M.R.; Demers, L.M.; Donahue, H.J.

    2003-01-01

    Disuse by bed rest, limb immobilization or space flight causes rapid bone loss by arresting bone formation and accelerating bone resorption. This net bone loss increases the risk of fracture upon remobilization. Bone loss also occurs in hibernating ground squirrels, golden hamsters, and little brown bats by arresting bone formation and accelerating bone resorption. There is some histological evidence to suggest that black bears Ursus americanus do not lose bone mass during hibernation (i.e. disuse). There is also evidence suggesting that muscle mass and strength are preserved in black bears during hibernation. The question of whether bears can prevent bone loss during hibernation has not been conclusively answered. The goal of the current study was to further assess bone metabolism in hibernating black bears. Using the same serum markers of bone remodeling used to evaluate human patients with osteoporosis, we assayed serum from five black bears, collected every 10 days over a 196-day period, for bone resorption and formation markers. Here we show that bone resorption remains elevated over the entire hibernation period compared to the pre-hibernation period, but osteoblastic bone formation is not impaired by hibernation and is rapidly accelerated during remobilization following hibernation.

  9. Temperature and acid-base balance in the American lobster Homarus americanus.

    PubMed

    Qadri, Syed Aman; Camacho, Joseph; Wang, Hongkun; Taylor, Josi R; Grosell, Martin; Worden, Mary Kate

    2007-04-01

    Lobsters (Homarus americanus) in the wild inhabit ocean waters where temperature can vary over a broad range (0-25 degrees C). To examine how environmental thermal variability might affect lobster physiology, we examine the effects of temperature and thermal change on the acid-base status of the lobster hemolymph. Total CO(2), pH, P(CO)2 and HCO(-)(3) were measured in hemolymph sampled from lobsters acclimated to temperature in the laboratory as well as from lobsters acclimated to seasonal temperatures in the wild. Our results demonstrate that the change in hemolymph pH as a function of temperature follows the rule of constant relative alkalinity in lobsters acclimated to temperature over a period of weeks. However, thermal change can alter lobster acid-base status over a time course of minutes. Acute increases in temperature trigger a respiratory compensated metabolic acidosis of the hemolymph. Both the strength and frequency of the lobster heartbeat in vitro are modulated by changes in pH within the physiological range measured in vivo. These observations suggest that changes in acid-base status triggered by thermal variations in the environment might modulate lobster cardiac performance in vivo.

  10. Sarcocystis oreamni, n. sp. (Apicomplexa: Sarcocystidae) from the mountain goat (Oreamnos americanus).

    PubMed

    Calero-Bernal, Rafael; Van Wilpe, Erna; White, Kevin; Verma, Shiv K; Cerqueira-Cézar, Camila K; Dubey, Jitender P

    2015-11-01

    Numerous species of Sarcocystis have been reported from wild ruminants, but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of three of seven mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; one had Sarcocystis cornagliai-like sarcocysts, previously named from the Alpine ibex (Capra ibex) from Europe. Two other goats were infected with a new species, Sarcocystis oreamni. Sarcocystis oreamni sarcocysts were microscopic with 2 μm-thick sarcocyst wall. By transmission electron microscopy, the sarcocyst wall had 1.7 μm-thick with unusual molar tooth-like villar protrusions (vp), type 29. The vp had an electron dense core and two disc-shaped plaques at the tip with fine microtubules. Bradyzoites were 8.6-9.1 μm long. Single nucleotide polymorphism (SNP) identified in 18S rRNA, and 28S rRNA loci of rDNA regions that suggested S. oreamni molecularly apart from related species. The phylogenetic analysis based on 18S rRNA and 28S rRNA sequences suggested S. oreamni is related with Sarcocystis species that employ members of the Canidae family as their definitive host. PMID:26255900

  11. Temporal dynamics of genetic variability in a mountain goat (Oreamnos americanus) population.

    PubMed

    Ortego, Joaquín; Yannic, Glenn; Shafer, Aaron B A; Mainguy, Julien; Festa-Bianchet, Marco; Coltman, David W; Côté, Steeve D

    2011-04-01

    The association between population dynamics and genetic variability is of fundamental importance for both evolutionary and conservation biology. We combined long-term population monitoring and molecular genetic data from 123 offspring and their parents at 28 microsatellite loci to investigate changes in genetic diversity over 14 cohorts in a small and relatively isolated population of mountain goats (Oreamnos americanus) during a period of demographic increase. Offspring heterozygosity decreased while parental genetic similarity and inbreeding coefficients (F(IS) ) increased over the study period (1995-2008). Immigrants introduced three novel alleles into the population and matings between residents and immigrants produced more heterozygous offspring than local crosses, suggesting that immigration can increase population genetic variability. The population experienced genetic drift over the study period, reflected by a reduced allelic richness over time and an 'isolation-by-time' pattern of genetic structure. The temporal decline of individual genetic diversity despite increasing population size probably resulted from a combination of genetic drift due to small effective population size, inbreeding and insufficient counterbalancing by immigration. This study highlights the importance of long-term genetic monitoring to understand how demographic processes influence temporal changes of genetic diversity in long-lived organisms. PMID:21366746

  12. American Lobsters (Homarus Americanus) not Surviving During Air Transport: Evaluation of Microbial Spoilage

    PubMed Central

    Tirloni, Erica; Stella, Simone; Gennari, Mario; Colombo, Fabio

    2016-01-01

    Eighteen American lobsters (Homarus americanus), dead during air transport, were analysed in order to evaluate the microbial population of meat, gills and gut: no specific studies have ever been conducted so far on the microbiological quality of American lobsters’ meats in terms of spoilage microbiota. The meat samples showed very limited total viable counts, in almost all the cases below the level of 6 Log CFU/g, while higher loads were found, as expected, in gut and gills, the most probable source of contamination. These data could justify the possibility to commercialise these not-surviving subjects, without quality concerns for the consumers. Most of the isolates resulted to be clustered with type strains of Pseudoalteromonas spp. (43.1%) and Photobacterium spp. (24.1%), and in particular to species related to the natural marine environment. The distribution of the genera showed a marked inhomogeneity among the samples. The majority of the isolates identified resulted to possess proteolytic (69.3%) and lipolytic ability (75.5%), suggesting their potential spoilage ability. The maintanance of good hygienical practices, especially during the production of ready-to-eat lobsters-based products, and a proper storage could limit the possible replication of these microorganisms. PMID:27800442

  13. Behavior and substrate selection during larval settling in the lobster Homarus americanus

    SciTech Connect

    Botero, L.; Atema, J.

    1982-02-01

    During the molt from third- to fourth-stage, larvae of Homarus americanus metamorphose into their adult form and pigmentation. In the course of the fourth stage, their pelagic life changes to a benthic existence. Artificial substrate choice experiments and qualitative illumination experiments show that during the early fourth stage the phototactic response reverses from positive to negative. Together with positive thigmotaxis, this results in a choice of dark crevices. Among natural substrate choices, preferential settlement occurred on macroalgal-covered rocks, followed by rocks on sand, mud, and sand. Although mud was not a preferred substrate in choice tests, the animals that chose mud and those that were presented only with mud settled successfully and were immediately efficient in their burrowing behavior, constructing U-shaped tunnels even in the center of the aquarium without using a pebble or rock as a starting point. These laboratory test confirm field observations that lobsters can successfully exploit a variety of substrates. They show that a substrate with preformed crevices is preferred for settling, but other substrates can be manipulated to make suitable burrows.

  14. Electrogenic sulfate uptake by crustacean hepatopancreatic basolateral membrane vesicles. [Homarus americanus

    SciTech Connect

    Cattey, M.A.; Gerencser, G.A.; Aheam, G.A. Univ. of Florida, Gainesville )

    1990-02-26

    Basolateral membrane vesicles (BLMV) were isolated from Atlantic lobster (Homarus americanus) hepatopancreas and purified by discontinuous sucrose gradient centrifugation. BLMV prepared in this fashion were osmotically reactive exhibiting linear dependence of vesicular {sup 35}SO{sub 4}{sup {minus}2} uptake to increasing external osmotic pressure with negligible non-specific isotope binding. Under short circuited conditions (valinomycin/K{sup +}) BLMV responded to either a HCO{sub 3}{sup {minus}} gradient directed out or equilibrated HCO{sub 3}{sup {minus}} (10 mM) by displaying short term accumulation of sulfate above that of equilibrium. Uptake of divalent anion was unaffected by an inwardly directed transmembrane Na{sup +} or tetramethylammonium{sup +} gradient. {sup 35}SO{sub 4}{sup {minus}2}/HCO{sub 3}{sup {minus}} exchange in the presence of valinomycin was stimulated by transient inside positive K{sup +} diffusion potentials and inhibited by transient inside negative K{sup +} diffusion potentials. The role of electrogenic anion exchange by hepatopancreas BLMV in transcellular sulfate transport is discussed.

  15. Sulfate uptake by crustacean hepatopancreatic brush border membrane vesicles. [Homarus americanus

    SciTech Connect

    Gerencser, G.A.; Cattey, M.A; Ahearn, G.A. Univ. of Hawaii, Honolulu )

    1990-02-26

    Purified brush border membrane vesicles (BBMV) were prepared from Atlantic lobster (Homarus americanus) hepatopancreas using differential centrifugation and Mg{sup +2} precipitation techniques. Uptake of 0.1 mM {sup 35}SO{sub 4}{sup {minus}2} was stimulated by pre-loading vesicles with Cl{sup {minus}} leading to a transient accumulation of isotope more than twice that at equilibrium. Pre-loading with HCO{sub 3}{sup {minus}} or gluconate had no effect on sulfate uptake. No stimulation of {sup 35}SO{sub 4}{sup {minus}2} was observed in the presence of inwardly directed Na{sup +} or tetramethylammonium{sup +} gradients. Uptake of the divalent anion was strongly stimulated by inwardly directed proton gradients (pH{sub o} < pH{sub i}) and markedly inhibited by outwardly directed proton gradients (pH{sub o} > pH{sub i}). {sup 35}SO{sub 4}{sup {minus}2}/Cl{sup {minus}} exchange was enhanced by imposing a transmembrane inside positive K{sup +} diffusion potential and inhibited by a membrane potential of the opposite polarity (K{sup +}/valinomycin). Results suggest the presence of a proton-dependent, electrogenic anion antiport mechanism in BBMV isolated from the crustacean hepatopancreas.

  16. Daily Rhythms of PERIOD protein in the eyestalk of the American lobster, Homarus americanus.

    PubMed

    Grabek, Katharine R; Chabot, Christopher C

    2012-01-01

    The daily rhythm of PERIOD protein (PER) expression is an integral component of the circadian clock, which is found among a broad range of animal species including fruit flies, marine mollusks and even humans. The use of antibodies directed against PER has provided a helpful tool in the discovery of PER homologues and the labeling of putative pacemaker cells, especially in animals for which an annotated genome is not readily available. In this study, DrosophilaPER antibodies were used to probe for PER in the American lobster, Homarus americanus. This species exhibits robust endogenous circadian rhythms but the circadian clock has yet to be located or characterized. PER was detected in the eyestalks of the lobster but not in the brain. Furthermore, a significant effect of the LD cycle on daily PER abundance was identified, and PER was significantly more abundant at mid dark than in early light or mid light hours. Our results suggest that PER is a part of the molecular machinery of the circadian clock located in the eyestalk of the lobster.

  17. An unusual cuticular tumor-like growth on the abdomen of a lobster, Homarus americanus.

    PubMed

    Shields, Jeffrey D; Small, Hamish J

    2013-11-01

    Tumors are rare in crustaceans, and whereas a few have been reported from the lobster Homarus americanus none have been adequately described. A lobster with an unusual, large, blue-colored tumor-like growth projecting laterally outward from the first abdominal somite was caught off Stonington, Maine, USA. The growth was rugose and covered by a relatively normal appearing cuticle with dispersed focal melanization. The underlying stroma consisted of an internal area of rescaffolded fibrous connective tissue, restructured muscle fibers, few arterioles, and an epidermal area comprised of columnar, highly vacuolated epithelial cells. No infectious pathogens or unusual inclusions were observed with microscopy and no eukaryotic pathogens were detected via molecular sequencing. Given the nature of the histology and the appearance of the growth, we identify the mass as a benign papilliform hamartoma that likely originated as a result of abnormal wound repair possibly initiated around ecdysis. This represents the first tumor-like hamartoma reported from a lobster, and the second hamartoma reported from a crustacean.

  18. Experimental infection and detection of necrotizing hepatopancreatitis bacterium in the American lobster Homarus americanus.

    PubMed

    Avila-Villa, Luz A; Gollas-Galván, Teresa; Martínez-Porchas, Marcel; Mendoza-Cano, Fernando; Hernández-López, Jorge

    2012-01-01

    Necrotizing hepatopancreatitis bacterium (NHPB) is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus) with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo.

  19. Antimicrobial activity in the cuticle of the American lobster, Homarus americanus.

    PubMed

    Mars Brisbin, Margaret; McElroy, Anne E; Pales Espinosa, Emmanuelle; Allam, Bassem

    2015-06-01

    American lobster, Homarus americanus, continues to be an ecologically and socioeconomically important species despite a severe decline in catches from Southern New England and Long Island Sound (USA) and a high prevalence of epizootic shell disease in these populations. A better understanding of lobster immune defenses remains necessary. Cuticle material collected from Long Island Sound lobsters was found to be active against a broad spectrum of bacteria, including Gram-negative and -positive species. The antimicrobial activity was characterized by boiling, muffling, and size fractioning. Boiling did not significantly reduce activity, while muffling did have a significant effect, suggesting that the active component is organic and heat stable. Size fractioning with 3 and 10 kDa filters did not significantly affect activity. Fast protein liquid chromatography fractions were also tested for antimicrobial activity, and fractions exhibiting protein peaks remained active. MALDI mass spectrometry revealed peptide peaks at 1.6, 2.8, 4.6, and 5.6 kDa. The data presented suggest that one or several antimicrobial peptides contribute to antimicrobial activity present in the American lobster cuticle.

  20. Anaerobic oral flora in the North American black bear (Ursus americanus) in eastern North Carolina.

    PubMed

    Clarke, Elsburgh O; Stoskopf, Michael K; Minter, Larry J; Stringer, Elizabeth M

    2012-06-01

    Microbial flora can provide insight into the ecology and natural history of wildlife in addition to improving understanding of health risks. This study examines the anaerobic oral flora of hunter killed black bears (Ursus americanus) in eastern North Carolina. Oral swabs from the buccal and lingual supragingival tooth surfaces of the first and second mandibular and maxillary molars of 22 black bears were inoculated onto Brucella Blood Agar plates supplemented with hemin and vitamin K after transport from the field using reduced oxoid nutrient broth. Sixteen anaerobic bacterial species, representing nine genera were identified using the RapID ANA II Micromethod Kit system and a number of organisms grown that could not be identified with the system. The most frequently identified anaerobes were Peptostreptococcus prevotii, Streptococcus constellatus, and Porphyromonas gingivalis. The diversity in the anaerobic oral flora of black bear in eastern North Carolina suggests the importance of including these organisms in basic health risk assessment protocols and suggests a potential tool for assessment of bear/habitat interactions. PMID:22503889

  1. Serosurvey for selected pathogens in free-ranging American black bears (Ursus americanus) in Maryland, USA.

    PubMed

    Bronson, Ellen; Spiker, Harry; Driscoll, Cindy P

    2014-10-01

    American black bears (Ursus americanus) in Maryland, USA, live in forested areas in close proximity to humans and their domestic pets. From 1999 to 2011, we collected 84 serum samples from 63 black bears (18 males; 45 females) in five Maryland counties and tested them for exposure to infectious, including zoonotic, pathogens. A large portion of the bears had antibody to canine distemper virus and Toxoplasma gondii, many at high titers. Prevalences of antibodies to zoonotic agents such as rabies virus and to infectious agents of carnivores including canine adenovirus and canine parvovirus were lower. Bears also had antibodies to vector-borne pathogens common to bears and humans such as West Nile virus, Borrelia burgdorferi, Rickettsia rickettsii, and Anaplasma phagocytophilum. Antibodies were detected to Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Canicola, Grippotyphosa, and Bratislava. We did not detect antibodies to Brucella canis or Ehrlichia canis. Although this population of Maryland black bears demonstrated exposure to multiple pathogens of concern for humans and domesticated animals, the low levels of clinical disease in this and other free-ranging black bear populations indicate the black bear is likely a spillover host for the majority of pathogens studied. Nevertheless, bear populations living at the human-domestic-wildlife interface with increasing human and domestic animal exposure should continue to be monitored because this population likely serves as a useful sentinel of ecosystem health. PMID:25075540

  2. Bears "Count" Too: Quantity Estimation and Comparison in Black Bears (Ursus Americanus).

    PubMed

    Vonk, Jennifer; Beran, Michael J

    2012-07-01

    Studies of bear cognition are notably missing from the comparative record despite bears' large relative brain size and interesting status as generalist carnivores facing complex foraging challenges, but lacking complex social structures. We investigated the numerical abilities of three American black bears (Ursus Americanus) by presenting discrimination tasks on a touch-screen computer. One bear chose the larger of two arrays of dot stimuli, while two bears chose the smaller array of dots. On some trials the relative number of dots was congruent with the relative total area of the two arrays. On other trials number of dots was incongruent with area. All of the bears were above chance on trials of both types with static dots. Despite encountering greater difficulty with dots that moved within the arrays, one bear was able to discriminate numerically larger arrays of moving dots, and a subset of moving dots from within the larger array, even when area and number were incongruent. Thus, although the bears used area as a cue to guide responding, they were also able to use number as a cue. The pattern of performance was similar to that found previously with monkeys, and suggests that bears may also show other forms of sophisticated quantitative abilities. PMID:22822244

  3. Temporal dynamics of genetic variability in a mountain goat (Oreamnos americanus) population.

    PubMed

    Ortego, Joaquín; Yannic, Glenn; Shafer, Aaron B A; Mainguy, Julien; Festa-Bianchet, Marco; Coltman, David W; Côté, Steeve D

    2011-04-01

    The association between population dynamics and genetic variability is of fundamental importance for both evolutionary and conservation biology. We combined long-term population monitoring and molecular genetic data from 123 offspring and their parents at 28 microsatellite loci to investigate changes in genetic diversity over 14 cohorts in a small and relatively isolated population of mountain goats (Oreamnos americanus) during a period of demographic increase. Offspring heterozygosity decreased while parental genetic similarity and inbreeding coefficients (F(IS) ) increased over the study period (1995-2008). Immigrants introduced three novel alleles into the population and matings between residents and immigrants produced more heterozygous offspring than local crosses, suggesting that immigration can increase population genetic variability. The population experienced genetic drift over the study period, reflected by a reduced allelic richness over time and an 'isolation-by-time' pattern of genetic structure. The temporal decline of individual genetic diversity despite increasing population size probably resulted from a combination of genetic drift due to small effective population size, inbreeding and insufficient counterbalancing by immigration. This study highlights the importance of long-term genetic monitoring to understand how demographic processes influence temporal changes of genetic diversity in long-lived organisms.

  4. Sarcocystis oreamni, n. sp. (Apicomplexa: Sarcocystidae) from the mountain goat (Oreamnos americanus).

    PubMed

    Calero-Bernal, Rafael; Van Wilpe, Erna; White, Kevin; Verma, Shiv K; Cerqueira-Cézar, Camila K; Dubey, Jitender P

    2015-11-01

    Numerous species of Sarcocystis have been reported from wild ruminants, but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of three of seven mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; one had Sarcocystis cornagliai-like sarcocysts, previously named from the Alpine ibex (Capra ibex) from Europe. Two other goats were infected with a new species, Sarcocystis oreamni. Sarcocystis oreamni sarcocysts were microscopic with 2 μm-thick sarcocyst wall. By transmission electron microscopy, the sarcocyst wall had 1.7 μm-thick with unusual molar tooth-like villar protrusions (vp), type 29. The vp had an electron dense core and two disc-shaped plaques at the tip with fine microtubules. Bradyzoites were 8.6-9.1 μm long. Single nucleotide polymorphism (SNP) identified in 18S rRNA, and 28S rRNA loci of rDNA regions that suggested S. oreamni molecularly apart from related species. The phylogenetic analysis based on 18S rRNA and 28S rRNA sequences suggested S. oreamni is related with Sarcocystis species that employ members of the Canidae family as their definitive host.

  5. Energy expenditure during activity in the American lobster Homarus americanus: Correlations with body acceleration.

    PubMed

    Lyons, G N; Halsey, L G; Pope, E C; Eddington, J D; Houghton, J D R

    2013-10-01

    How animals manage time and expend energy has implications for survivorship. Being able to measure key metabolic costs of animals under natural conditions is therefore an important tool in behavioral ecology. One method for estimating activity-specific metabolic rate is via derived measures of acceleration, often 'overall dynamic body acceleration' (ODBA), recorded by an instrumented acceleration logger. ODBA has been shown to correlate well with rate of oxygen consumption (V˙o2) in a range of species during activity in the laboratory. This study devised a method for attaching acceleration loggers to decapod crustaceans and then correlated ODBA against concurrent respirometry readings to assess accelerometry as a proxy for activity-specific energy expenditure in a model species, the American lobster Homarus americanus. Where the instrumented animals exhibited a sufficient range of activity levels, positive linear relationships were found between V˙o2 and ODBA over 20min periods at a range of ambient temperatures (6, 13 and 20°C). Mixed effect linear models based on these data and morphometrics provided reasonably strong predictive power for estimating activity-specific V˙o2 from ODBA. These V˙o2-ODBA calibrations demonstrate the potential of accelerometry as an effective predictor of behavior-specific metabolic rate of crustaceans in the wild during periods of activity.

  6. Bears “Count” Too: Quantity Estimation and Comparison in Black Bears (Ursus Americanus)

    PubMed Central

    Vonk, Jennifer; Beran, Michael J.

    2012-01-01

    Studies of bear cognition are notably missing from the comparative record despite bears’ large relative brain size and interesting status as generalist carnivores facing complex foraging challenges, but lacking complex social structures. We investigated the numerical abilities of three American black bears (Ursus Americanus) by presenting discrimination tasks on a touch-screen computer. One bear chose the larger of two arrays of dot stimuli, while two bears chose the smaller array of dots. On some trials the relative number of dots was congruent with the relative total area of the two arrays. On other trials number of dots was incongruent with area. All of the bears were above chance on trials of both types with static dots. Despite encountering greater difficulty with dots that moved within the arrays, one bear was able to discriminate numerically larger arrays of moving dots, and a subset of moving dots from within the larger array, even when area and number were incongruent. Thus, although the bears used area as a cue to guide responding, they were also able to use number as a cue. The pattern of performance was similar to that found previously with monkeys, and suggests that bears may also show other forms of sophisticated quantitative abilities. PMID:22822244

  7. Behavioral response and kinetics of terrestrial atrazine exposure in American toads (bufo americanus)

    USGS Publications Warehouse

    Storrs, Mendez S.I.; Tillitt, D.E.; Rittenhouse, T.A.G.; Semlitsch, R.D.

    2009-01-01

    Amphibians in terrestrial environments obtain water through a highly vascularized pelvic patch of skin. Chemicals can also be exchanged across this patch. Atrazine (ATZ), a widespread herbicide, continues to be a concern among amphibian ecologists based on potential exposure and toxicity. Few studies have examined its impact on the terrestrial juvenile or adult stages of toads. In the current study, we asked the following questions: (1) Will juvenile American toads (Bufo americanus) avoid soils contaminated with ATZ? (2) Can they absorb ATZ across the pelvic patch? (3) If so, how is it distributed among the organs and eventually eliminated? We conducted a behavioral choice test between control soil and soil dosed with ecologically relevant concentrations of ATZ. In addition, we examined the uptake, distribution, and elimination of water dosed with 14C-labeled ATZ. Our data demonstrate that toads do not avoid ATZ-laden soils. ATZ crossed the pelvic patch rapidly and reached an apparent equilibrium within 5 h. The majority of the radiolabeled ATZ ended up in the intestines, whereas the greatest concentrations were observed in the gall bladder. Thus, exposure of adult life stages of amphibians through direct uptake of ATZ from soils and runoff water should be considered in risk evaluations. ?? 2009 Springer Science+Business Media, LLC.

  8. Antimicrobial activity in the cuticle of the American lobster, Homarus americanus.

    PubMed

    Mars Brisbin, Margaret; McElroy, Anne E; Pales Espinosa, Emmanuelle; Allam, Bassem

    2015-06-01

    American lobster, Homarus americanus, continues to be an ecologically and socioeconomically important species despite a severe decline in catches from Southern New England and Long Island Sound (USA) and a high prevalence of epizootic shell disease in these populations. A better understanding of lobster immune defenses remains necessary. Cuticle material collected from Long Island Sound lobsters was found to be active against a broad spectrum of bacteria, including Gram-negative and -positive species. The antimicrobial activity was characterized by boiling, muffling, and size fractioning. Boiling did not significantly reduce activity, while muffling did have a significant effect, suggesting that the active component is organic and heat stable. Size fractioning with 3 and 10 kDa filters did not significantly affect activity. Fast protein liquid chromatography fractions were also tested for antimicrobial activity, and fractions exhibiting protein peaks remained active. MALDI mass spectrometry revealed peptide peaks at 1.6, 2.8, 4.6, and 5.6 kDa. The data presented suggest that one or several antimicrobial peptides contribute to antimicrobial activity present in the American lobster cuticle. PMID:25804485

  9. Social network analysis of mating patterns in American black bears (Ursus americanus).

    PubMed

    Moore, Jennifer A; Xu, Ran; Frank, Kenneth; Draheim, Hope; Scribner, Kim T

    2015-08-01

    Nonrandom mating can structure populations and has important implications for population-level processes. Investigating how and why mating deviates from random is important for understanding evolutionary processes as well as informing conservation and management. Prior to the implementation of parentage analyses, understanding mating patterns in solitary, elusive species like bears was virtually impossible. Here, we capitalize on a long-term genetic data set collected from black bears (Ursus americanus) (N = 2422) in the Northern Lower Peninsula (NLP) of Michigan, USA. We identified mated pairs using parentage analysis and applied logistic regression (selection) models that controlled for features of the social network, to quantify the effects of individual characteristics, and spatial and population demographic factors on mating dynamics. Logistic regression models revealed that black bear mating was associated with spatial proximity of mates, male age, the time a pair had coexisted, local population density and relatedness. Mated pairs were more likely to contain older males. On average, bears tended to mate with nearby individuals to whom they were related, which does not support the existence of kin recognition in black bears. Pairwise relatedness was especially high for mated pairs containing young males. Restricted dispersal and high male turnover from intensive harvest mortality of NLP black bears are probably the underlying factors associated with younger male bears mating more often with female relatives. Our findings illustrate how harvest has the potential to disrupt the social structure of game species, which warrants further attention for conservation and management.

  10. Necator americanus secretory acetylcholinesterase and its purification from excretory-secretory products by affinity chromatography.

    PubMed

    Pritchard, D I; Leggett, K V; Rogan, M T; McKean, P G; Brown, A

    1991-03-01

    Acetylcholinesterase (AChE) secretion by adult N. americanus was enhanced in vitro by incorporating insoluble collagen rafts into culture dishes. Enzyme produced in this way had preferential substrate specificity for acetylthiocholine iodide (ATC), and its activity was inhibited by eserine (1.1 x 10(-8) M). Ancylostoma ceylanicum, another hookworm species, failed to produce comparable amounts of AChE in culture. AChE was efficiently purified from culture medium by affinity chromatography on edrophonium sepharose; 81% of the AChE activity was retained by the affinity matrix, although this fraction contained only 4.3% of the protein loaded. Antisera raised against purified AChE in rabbits immunohistochemically stained the oesophageal glands of the parasite, and reacted with molecules of 32, 60, 80, 140 and 220 kDa in reduced adult ES products on Western blotting, although differential activity was observed against worm homogenates and earlier developmental stages. On IEF, purified AChE resolved predominantly with a pl of 3.55; proteins with a similar pl were recognized by rabbit anti-AChE. IgG preparations of this antiserum inhibited AChE activity in ES products, and inhibited AChE secretion by adult worms in culture. The availability of this immunological probe will allow definitive experiments to be conducted on the role of this enigmatic enzyme in the host-parasite relationship. PMID:2052405

  11. Influence of long (16L:8D) and short (8L:16D) photoperiods on blood metabolites and hepatic metabolism in Olive flounder, Paralichthys olivaceus.

    PubMed

    Zou, Huafeng; Bai, Xianshou; Feng, Yuhong; Zhang, Ying; Wang, Youji; Lu, Weiqun

    2016-01-01

    In the present study the influence of long photoperiod (LP, 16L:8D) and short photoperiod (SP, 8L:16D) on hepatic energy metabolism in the olive flounder (Paralichthys olivaceus) was investigated. Flounders were maintained under LP or SP conditions for 2 weeks then plasmatic and hepatic parameters were assessed. At the plasmatic level, the concentration of cortisol was enhanced in flounder maintained under LP compared to SP. Alkaline phosphatase (ALP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzyme activities in plasma also increased in LP flounder. There was no significant difference in plasma glucose levels between the two experimental groups. Plasma osmotic pressure, Na and Cl levels were significantly higher in LP compared to the SP group. In liver, a significant decrease of triglycerides together with an increase in glycogen was observed in the LP group. Hepatic hsl and pepck and muscle hsl mRNA expression in LP was significantly higher in the SP group. Overall the results indicate that the LP treatment caused a mild stress response and increased hepatic energy metabolism in the flounder, which in turn could affect osmoregulation. In conclusion, it would appear that LP treatment can adversely influence hepatic energy metabolism in adult olive flounder under fasting condition. PMID:27386368

  12. Molecular and functional characterization of opsins in barfin flounder (Verasper moseri).

    PubMed

    Kasagi, Satoshi; Mizusawa, Kanta; Murakami, Naoto; Andoh, Tadashi; Furufuji, Sumihisa; Kawamura, Shoji; Takahashi, Akiyoshi

    2015-02-10

    Green light irradiation facilitates the somatic growth of barfin flounder (Verasper moseri). However, the V. moseri visual system, which may be associated with somatic growth by acting on the endocrine system upon exposure to this particular wavelength, remains largely unexplored. Herein, we characterized the visual opsin repertoire of V. moseri to understand the molecular basis underlying this effect. The five types of visual opsins that are found in vertebrates were cloned from RNA that was extracted from the eyes of V. moseri. Notably, V. moseri possessed one pseudogene (RH2-A) and two intact (RH2-B and RH2-C) copies of "green-sensitive" opsin genes. The wavelengths of maximum absorption spectra (λmax) for each of the reconstituted photopigments were 552nm for "red-sensitive" LWS, 506nm for RH2-B, 490nm for RH2-C, 482nm and 416nm for "blue-sensitive" SWS2A and SWS2B, respectively, 367nm for "ultraviolet-sensitive" SWS1, and 494nm for "dim-light sensitive rhodopsin" RH1. The λmax of SWS2A was longer than that of any other reported vertebrate SWS2 opsin. By measuring the expression level of these opsin genes with quantitative RT-PCR in 3-, 15-, and 27-month-old fish, we found that RH2-B and SWS2A were expressed at a constant level, whereas the expression of LWS, RH2-C, SWS2B, and SWS1 opsin genes decreased, and that of RH1 increased with age. Barfin flounders inhabit inshore waters at a young age and expand their habitat to deep sea areas as they age, and green light is relatively abundant in deep water compared to the lights of other wavelengths in shallow water. Our results indicate that gene repertoire and expression profile of the opsin genes of barfin flounder are adaptive to their habitat shift that occurs during development, with some opsins acquiring a distinct λmax.

  13. Differential expression of American lobster (Homarus americanus) immune related genes during infection of Aerococcus viridans var. homari, the causative agent of Gaffkemia.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Greenwood, Spencer J

    2013-02-01

    This is the first transcriptomic study focusing on immunity in the commercially valuable American lobster (Homarus americanus). We have conducted an in vivo infection trial using the Gram-positive bacterium Aerococcus viridans var. homari to determine how H. americanus responds to this naturally occurring lethal-pathogen. A novel H. americanus microarray was used to measure the transcriptomic changes occurring in over 14,000 genes in the lobster hepatopancreas. Hundreds of new immune genes and isoforms were identified and measured for the first time in this species, and our findings highlight 148 genes of interest involved in H. americanus pathogen response. We verified our microarray results using RT-qPCR on three anti-lipopolysaccharide (ALFHa-1, ALFHa-2, ALFHa-4), a thioredoxin, acute phase serum amyloid protein A, hexokinase and two trypsin genes. RT-qPCR and microarray findings show close agreement and highlight the significant increase in gene expression in many lobster immune genes during A. viridans infection. Differential expression of the ALFHa isoforms may indicate that the H. americanus immune response can be tailored to the class of pathogen causing disease.

  14. Identification of candidate piRNAs in the gonads of Paralichthys olivaceus (Japanese flounder)

    PubMed Central

    WANG, Chun-Lei; WANG, Zhi-Peng; WANG, Jia-Qi; Li, Ming-You; CHEN, Xiao-Wu

    2016-01-01

    Piwi-interacting RNA (piRNA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, piRNA libraries derived from the adult gonads of Japanese flounder (Paralichthys olivaceus) were generated using next-generation sequencing technology. Using zebrafish piRNAs as a reference, 5 865 unique candidate piRNAs were identified; 289 candidate piRNA clusters (PRCs) were generated from the above piRNAs. Among the isolated candidate PRCs, a total of 38 ovary-specific, 45 ovary-bias, 24 testis-specific, and 131 testis-bias PRCs were found. The relative expression levels of seven PRCs were validated through quantitative reverse transcription-polymerase chain reaction. The results of this study will help facilitate exploration of the development and maintenance of the phenotypic sex mechanism in P. olivaceus. PMID:27686790

  15. Feeding and growth of juvenile Japanese flounder Paralichthys olivaceus in relation to temperature and food supply

    NASA Astrophysics Data System (ADS)

    Fonds, Mark; Tanaka, Masaru; Van der Veer, Henk W.

    The growth of juvenile Japanese flounder Paralichthys olivaceus (Temming et Schlegel) exposed to various feeding and temperature regimes was studied in the laboratory. The temperature limits for growth were estimated from about 9 to 32°C, and highest growth rate was observed at a temperature of 25°C. For fish of an initial total length of about 3 cm, exposed to temperatures between 10 and 30°C with unlimited food, the daily growth rate in length (dL; mm·d -1) was described as a function of temperature (T,°C) by the equation: dL=0.005 T 2 - 0.0000046 T 4 - 0.35. Fish fed with mussel meat or mysids displayed similar protein conversion efficiencies. The relationship between condition factor of the fish and the proximate body composition of the fish is reported.

  16. Identification of candidate piRNAs in the gonads of Paralichthys olivaceus (Japanese flounder).

    PubMed

    Wang, Chun-Lei; Wang, Zhi-Peng; Wang, Jia-Qi; Li, Ming-You; Chen, Xiao-Wu

    2016-09-18

    Piwi-interacting RNA (piRNA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, piRNA libraries derived from the adult gonads of Japanese flounder (Paralichthys olivaceus) were generated using next-generation sequencing technology. Using zebrafish piRNAs as a reference, 5 865 unique candidate piRNAs were identified; 289 candidate piRNA clusters (PRCs) were generated from the above piRNAs. Among the isolated candidate PRCs, a total of 38 ovary-specific, 45 ovary-bias, 24 testis-specific, and 131 testis-bias PRCs were found. The relative expression levels of seven PRCs were validated through quantitative reverse transcription-polymerase chain reaction. The results of this study will help facilitate exploration of the development and maintenance of the phenotypic sex mechanism in P. olivaceus. PMID:27686790

  17. Analysis of new microsatellite markers developed from reported sequences of Japanese flounder Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Yu, Haiyang; Jiang, Liming; Chen, Wei; Wang, Xubo; Wang, Zhigang; Zhang, Quanqi

    2010-12-01

    The expressed sequence tags (ESTs) of Japanese flounder, Paralichthys olivaceus, were selected from GenBank to identify simple sequence repeats (SSRs) or microsatellites. A bioinformatic analysis of 11111 ESTs identified 751 SSR-containing ESTs, including 440 dinucleotide, 254 trinucleotide, 53 tetranucleotide, 95 pentanucleotide and 40 hexanucleotide microsatellites respectively. The CA/TG and GA/TC repeats were the most abundant microsatellites. AT-rich types were predominant among trinucleotide and tetranucleotide microsatellites. PCR primers were designed to amplify 10 identified microsatellites loci. The PCR results from eight pairs of primers showed polymorphisms in wild populations. In 30 wild individuals, the mean observed and expected heterozygosities of these 8 polymorphic SSRs were 0.71 and 0.83 respectively and the average PIC value was 0.8. These microsatellite markers should prove to be a useful addition to the microsatellite markers that are now available for this species.

  18. Effects of resveratrol on growth and skeletal muscle physiology of juvenile southern flounder.

    PubMed

    Wilson, Whitney N; Baumgarner, Bradley L; Watanabe, Wade O; Alam, Md Shah; Kinsey, Stephen T

    2015-05-01

    Resveratrol is a naturally occurring antioxidant that has been widely studied in mammals due to its potential to extend lifespan. However, antioxidants may also limit protein damage and therefore reduce rates of protein degradation, providing a potential avenue for enhancing growth in an aquaculture setting. The present study tested the hypotheses that in Southern flounder, Paralichthys lethostigma, resveratrol would decrease protein carbonylation and 4-HNE (indicators of protein and lipid oxidative damage, respectively), levels of ubiquitinylation and LC3 (indicators of non-lysosomal and lysosomal protein degradation, respectively), while having no effect on S6K activation (indicator of protein synthesis). These effects were predicted to increase growth rate. Mitochondrial volume density was also examined since resveratrol may lead to the proliferation of mitochondria, which are the principal source of reactive oxygen species (ROS) that cause oxidative damage. Juvenile fish (n=142) were fed a control diet or a diet supplemented with 600 μg resveratrol per g of food for 16 weeks. Fish treated with resveratrol had a 9% greater length and 33% greater body mass than control fish after 16 weeks. Additionally, there was lower protein carbonylation and lipid 4-HNE within the muscle tissues of treated fish, indicating decreased oxidative damage, and reduced protein ubiquitinylation in the resveratrol fed flounder, indicating less protein degradation. However, there was not a significant difference in LC3, S6K activation, or mitochondrial volume density. These results suggest that resveratrol has positive effects on growth due to its antioxidant properties that reduce non-lysosomal protein degradation. PMID:25522711

  19. Effects of resveratrol on growth and skeletal muscle physiology of juvenile southern flounder.

    PubMed

    Wilson, Whitney N; Baumgarner, Bradley L; Watanabe, Wade O; Alam, Md Shah; Kinsey, Stephen T

    2015-05-01

    Resveratrol is a naturally occurring antioxidant that has been widely studied in mammals due to its potential to extend lifespan. However, antioxidants may also limit protein damage and therefore reduce rates of protein degradation, providing a potential avenue for enhancing growth in an aquaculture setting. The present study tested the hypotheses that in Southern flounder, Paralichthys lethostigma, resveratrol would decrease protein carbonylation and 4-HNE (indicators of protein and lipid oxidative damage, respectively), levels of ubiquitinylation and LC3 (indicators of non-lysosomal and lysosomal protein degradation, respectively), while having no effect on S6K activation (indicator of protein synthesis). These effects were predicted to increase growth rate. Mitochondrial volume density was also examined since resveratrol may lead to the proliferation of mitochondria, which are the principal source of reactive oxygen species (ROS) that cause oxidative damage. Juvenile fish (n=142) were fed a control diet or a diet supplemented with 600 μg resveratrol per g of food for 16 weeks. Fish treated with resveratrol had a 9% greater length and 33% greater body mass than control fish after 16 weeks. Additionally, there was lower protein carbonylation and lipid 4-HNE within the muscle tissues of treated fish, indicating decreased oxidative damage, and reduced protein ubiquitinylation in the resveratrol fed flounder, indicating less protein degradation. However, there was not a significant difference in LC3, S6K activation, or mitochondrial volume density. These results suggest that resveratrol has positive effects on growth due to its antioxidant properties that reduce non-lysosomal protein degradation.

  20. Seasonal variability of prey in the stomachs of Astropecten americanus (Echinodermata: Asteroidea) from off southern New England, U.S.A.

    NASA Astrophysics Data System (ADS)

    Franz, David R.; Worley, E. K.

    1982-04-01

    The diet of Astropecten americanus from a location off Nantucket Shoals, Massachusetts, was determined by analysis of stomach contents in May and November. Molluscs, followed by crustaceans, dominated stomach contents. The proportion of molluscan species in the diet differed significantly from the benthos, Juvenile Arctica islandica dominated stomachs in May, but declined sharply in November, probably due in part to predation by Astropecten. Other molluscs, not prevalent in the benthos, were important in stomach contents. Frequency distributions of counts of prey species in a sample of seastar stomachs indicate that some seastars ingest unusually large numbers of certain prey species. A. americanus generally ingests small prey individuals (<4mm) although larger prey are consumed, especially in November. Relatively high selectivity, seasonal shifts in the diversity of stomach contents, and evidence for patch-specific foraging are characteristics of A. americanus which are consistent with general predictions of optimum diet models.

  1. Fatal Caprine arthritis encephalitis virus-like infection in 4 Rocky Mountain goats (Oreamnos americanus).

    PubMed

    Patton, Kristin M; Bildfell, Robert J; Anderson, Mark L; Cebra, Christopher K; Valentine, Beth A

    2012-03-01

    Over a 3.5-year period, 4 Rocky Mountain goats (Oreamnos americanus), housed at a single facility, developed clinical disease attributed to infection by Caprine arthritis encephalitis virus (CAEV). Ages ranged from 1 to 10 years. Three of the goats, a 1-year-old female, a 2-year-old male, and a 5-year-old male, had been fed raw domestic goat milk from a single source that was later found to have CAEV on the premises. The fourth animal, a 10-year-old male, had not ingested domestic goat milk but had been housed with the other 3 Rocky Mountain goats. All 4 animals had clinical signs of pneumonia prior to death. At necropsy, findings in lungs included marked diffuse interstitial pneumonia characterized histologically by severe lymphoplasmacytic infiltrates with massive alveolar proteinosis, interstitial fibrosis, and type II pneumocyte hyperplasia. One animal also developed left-sided hemiparesis, and locally extensive lymphoplasmacytic myeloencephalitis was present in the cranial cervical spinal cord. Two animals had joint effusions, as well as severe lymphoplasmacytic and ulcerative synovitis. Immunohistochemical staining of fixed sections of lung tissue from all 4 goats, as well as spinal cord in 1 affected animal, and synovium from 2 affected animals were positive for CAEV antigen. Serology testing for anti-CAEV antibodies was positive in the 2 goats tested. The cases suggest that Rocky Mountain goats are susceptible to naturally occurring CAEV infection, that CAEV from domestic goats can be transmitted to this species through infected milk and by horizontal transmission, and that viral infection can result in clinically severe multisystemic disease. PMID:22379056

  2. Multilocus heterozygosity, parental relatedness and individual fitness components in a wild mountain goat, Oreamnos americanus population.

    PubMed

    Mainguy, Julien; Côté, Steeve D; Coltman, David W

    2009-05-01

    Matings between relatives lead to a decrease in offspring genetic diversity which can reduce fitness, a phenomenon known as inbreeding depression. Because alpine ungulates generally live in small structured populations and often exhibit a polygynous mating system, they are susceptible to inbreeding. Here, we used marker-based measures of pairwise genetic relatedness and inbreeding to investigate the fitness consequences of matings between relatives in a long-term study population of mountain goats (Oreamnos americanus) at Caw Ridge, Alberta, Canada. We first assessed whether individuals avoided mating with kin by comparing actual and random mating pairs according to their estimated genetic relatedness, which was derived from 25 unlinked polymorphic microsatellite markers and reflected pedigree relatedness. We then examined whether individual multilocus heterozygosity H, used as a measure of inbreeding, was predicted by parental relatedness and associated with yearling survival and the annual probability of giving birth to a kid in adult females. Breeding pairs identified by genetic parentage analyses of offspring that survived to 1 year of age were less genetically related than expected under random matings. Parental relatedness was negatively correlated with offspring H, and more heterozygous yearlings had higher survival to 2 years of age. The probability of giving birth was not affected by H in adult females. Because kids that survived to yearling age were mainly produced by less genetically related parents, our results suggest that some individuals experienced inbreeding depression in early life. Future research will be required to quantify the levels of gene flow between different herds, and evaluate their effects on population genetic diversity and dynamics. PMID:19389162

  3. Sequential ovulation and fertility of polyoestrus in American black bears (Ursus americanus)

    PubMed Central

    Himelright, Brendan M.; Moore, Jenna M.; Gonzales, Ramona L.; Mendoza, Alejandra V.; Dye, Penny S.; Schuett, Randall J.; Durrant, Barbara S.; Read, Betsy A.; Spady, Thomas J.

    2014-01-01

    American black bears (Ursus americanus) are seasonally polyoestrous and exhibit delayed implantation, which may allow equal and independent fertility of recurrent oestruses of a mating season. We postulated that the luteal inactivity during delayed implantation allows bears to have sequential ovulation during a polyoestrous mating season such that each oestrus of a polyoestrous female will have equivalent fertility, and pregnancy would not preclude subsequent ovulation and superfetation. Controlled mating experiments were conducted on semi-free-ranging female American black bears during three mating seasons, wherein females were bred by different male cohorts in each oestrus. Behavioural observation, vulva score ranking, genetic paternity analysis, gross morphology of ovaries and microscopic morphology of diapaused embryos were used to evaluate the fertility of each subsequent oestrus in polyoestrous females. Oestrus duration, number of successful mounts and median vulva scores were similar between first and subsequent oestruses of the season. Polyoestrus occurred in 81.3% of oestrous females, with a 9.7 ± 5.5 day (mean ± SD) inter-oestrous interval. Sequential ovulation was documented in three polyoestrous females, including one that possessed both a corpus haemorrhagicum and a developed corpus luteum. Among polyoestrous dams, four of nine embryos were conceived in the first oestrus and five of nine in the second oestrus. These results indicate that each oestrus of polyoestrous females is capable of fertility, even if the female is already pregnant from a prior oestrus. Although superfetation was not directly observed in the present study, our results strongly suggest the potential of superfetation in the American black bear and provide novel insight into the complex behavioural and physiological breeding mechanisms of bears. Given that most endangered bear species share similar reproductive traits with American black bears, captive breeding programmes

  4. Behavioral Thermoregulation and Trade-Offs in Juvenile Lobster Homarus americanus.

    PubMed

    Nielsen, Travis V; McGaw, Iain J

    2016-02-01

    Water temperature influences the behavior and distribution patterns of both larval and adult American lobster Homarus americanus. However, very little is known about the responses of juvenile lobsters. The juvenile life stage is a critical period; high levels of mortality, combined with specific behavioral responses, can disconnect larval settlement from patterns of abundance of adults. We assessed behavioral thermoregulation in juvenile lobsters, and determined how thermal preferences can be altered by the presence of shelter and food. Juvenile lobsters avoided temperatures higher than 20 °C and lower than 8 °C, and had a mean temperature preference of 16.2 ± 1 °C. This preference was unaffected by prior acclimation, origin (laboratory-raised or wild), or size. When the animals were subjected to a temperature change (5-20 °C), activity rates peaked at 15 °C, and remained stable thereafter. Activity rates did not change when a shelter was added. The addition of food resulted in an increase in activity associated with food handling. When juvenile lobsters were offered a choice between temperature, shelter, and food, they always chose the environment with a shelter, even when it was in a thermally unfavorable temperature. Juveniles also spent more time in a thermally unfavorable environment when food was present; however, acquisition of a shelter was prioritized over food. Although juveniles had a similar thermal preference to adults, they are more vulnerable to predation; the innate shelter-seeking behavior of juveniles overrode their thermal preference. While temperature is an important environmental factor affecting the physiology, distribution, and growth of aquatic ectotherms, our findings suggest that trade-off behaviors occur in order to maintain optimal fitness and survival of the individual.

  5. Structural and Functional Characterization of a Multifunctional Alanine-Rich Peptide Analogue from Pleuronectes americanus

    PubMed Central

    Migliolo, Ludovico; Silva, Osmar N.; Silva, Paula A.; Costa, Maysa P.; Costa, Carolina R.; Nolasco, Diego O.; Barbosa, João A. R. G.; Silva, Maria R. R.; Bemquerer, Marcelo P.; Lima, Lidia M. P.; Romanos, Maria T. V.; Freitas, Sonia M.; Magalhães, Beatriz S.; Franco, Octavio L.

    2012-01-01

    Recently, defense peptides that are able to act against several targets have been characterized. The present work focuses on structural and functional evaluation of the peptide analogue Pa-MAP, previously isolated as an antifreeze peptide from Pleuronectes americanus. Pa-MAP showed activities against different targets such as tumoral cells in culture (CACO-2, MCF-7 and HCT-116), bacteria (Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 25923), viruses (HSV-1 and HSV-2) and fungi (Candida parapsilosis ATCC 22019, Trichophyton mentagrophytes (28d&E) and T. rubrum (327)). This peptide did not show toxicity against mammalian cells such as erythrocytes, Vero and RAW 264.7 cells. Molecular mechanism of action was related to hydrophobic residues, since only the terminal amino group is charged at pH 7 as confirmed by potentiometric titration. In order to shed some light on its structure-function relations, in vitro and in silico assays were carried out using circular dichroism and molecular dynamics. Furthermore, Pa-MAP showed partial unfolding of the peptide changes in a wide pH (3 to 11) and temperature (25 to 95°C) ranges, although it might not reach complete unfolding at 95°C, suggesting a high conformational stability. This peptide also showed a conformational transition with a partial α-helical fold in water and a full α-helical core in SDS and TFE environments. These results were corroborated by spectral data measured at 222 nm and by 50 ns dynamic simulation. In conclusion, data reported here show that Pa-MAP is a potential candidate for drug design against pathogenic microorganisms due to its structural stability and wide activity against a range of targets. PMID:23056574

  6. Oocyte recovery and maturation in the American black bear (Ursus americanus): a model for endangered ursids.

    PubMed

    Johnston, L A; Donoghue, A M; Igo, W; Simmons, L G; Wildt, D E; Rieffenberger, J

    1994-05-15

    A study was conducted to determine if meiotic maturation could be induced in ovarian oocytes of the American black bear (Ursus americanus), a model for gamete "rescue" techniques for endangered ursids. Ovaries obtained from 48 black bears yielded 2,403 oocytes (51.1 +/- 4.9/female), of which 777 (32.3%) were morphologically classified as excellent quality. More total oocytes were recovered from donors that were anestrous compared to luteal/pregnant (P < 0.05) at the time of ovarian excision. Delaying the recovery of oocytes from antral follicles within excised ovaries from 12-24 hr to 25-36 hr had no effect (P > 0.05) on the overall number of high quality oocytes recovered or subsequent maturational ability. The highest incidence of metaphase II was reached between 48 and 60 hr of in vitro incubation. Donor status (anestrous vs. luteal/pregnant) had no influence on the oocyte maturation rate by 24 or 48 hr, but by 60 hr, more (P < 0.05) oocytes recovered from anestrous females (43.9%) had achieved metaphase II compared to luteal/pregnant counterparts (23.1%). In preliminary trials involving endangered ursids, 54 ovarian oocytes were recovered from three aged sun bears (Helarctos malayanus), of which 72.2% were excellent quality and 15.4% matured in vitro to metaphase II. Similarly, 119 antral oocytes were recovered from two aged sloth bears (Melursus ursinus), of which 41.2% were excellent and 17.5% matured in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8207381

  7. Sequential ovulation and fertility of polyoestrus in American black bears (Ursus americanus).

    PubMed

    Himelright, Brendan M; Moore, Jenna M; Gonzales, Ramona L; Mendoza, Alejandra V; Dye, Penny S; Schuett, Randall J; Durrant, Barbara S; Read, Betsy A; Spady, Thomas J

    2014-01-01

    American black bears (Ursus americanus) are seasonally polyoestrous and exhibit delayed implantation, which may allow equal and independent fertility of recurrent oestruses of a mating season. We postulated that the luteal inactivity during delayed implantation allows bears to have sequential ovulation during a polyoestrous mating season such that each oestrus of a polyoestrous female will have equivalent fertility, and pregnancy would not preclude subsequent ovulation and superfetation. Controlled mating experiments were conducted on semi-free-ranging female American black bears during three mating seasons, wherein females were bred by different male cohorts in each oestrus. Behavioural observation, vulva score ranking, genetic paternity analysis, gross morphology of ovaries and microscopic morphology of diapaused embryos were used to evaluate the fertility of each subsequent oestrus in polyoestrous females. Oestrus duration, number of successful mounts and median vulva scores were similar between first and subsequent oestruses of the season. Polyoestrus occurred in 81.3% of oestrous females, with a 9.7 ± 5.5 day (mean ± SD) inter-oestrous interval. Sequential ovulation was documented in three polyoestrous females, including one that possessed both a corpus haemorrhagicum and a developed corpus luteum. Among polyoestrous dams, four of nine embryos were conceived in the first oestrus and five of nine in the second oestrus. These results indicate that each oestrus of polyoestrous females is capable of fertility, even if the female is already pregnant from a prior oestrus. Although superfetation was not directly observed in the present study, our results strongly suggest the potential of superfetation in the American black bear and provide novel insight into the complex behavioural and physiological breeding mechanisms of bears. Given that most endangered bear species share similar reproductive traits with American black bears, captive breeding programmes

  8. Thyroid cystadenoma, colloid goiter, and hypothyroidism in an American black bear (Ursus americanus).

    PubMed

    Storms, Timothy N; Beazley, Shelley L; Schumacher, Juergen; Ramsay, Edward C

    2004-03-01

    A 178-kg, 14-yr-old captive female American black bear (Ursus americanus) was examined because of lethargy, inappetance, obesity, and alopecia. Serum chemistry and complete blood count values were within normal limits. Based on serum levels for total thyroxine (T4), free T4 by equilibrium dialysis (fT4ED), and canine thyroid-stimulating hormone concentrations, using assays validated for domestic dogs, hypothyroidism was diagnosed presumptively, and therapy with levothyroxine sodium (0.022 mg/kg p.o. b.i.d.) was initiated. Haircoat, body weight, appetite, and activity level improved within 30 days. The levothyroxine dose was decreased twice (to 0.018 mg/kg p.o. b.i.d. and then to 0.011 mg/kg p.o. b.i.d.) during the course of treatment based on monitoring of serum T4 and fT4ED concentrations. After euthanasia for severe refractory lameness, postmortem examination revealed bilateral thyroid lobe enlargement and a fluid-filled cyst within the right lobe. Histologically, colloid goiter was present in both lobes, and a follicular cystadenoma had replaced one third of the cranial pole of the right lobe. The goiter and cystadenoma likely contributed to the hypothyroid condition in this bear and fT4ED was a more sensitive indicator of hypothyroidism than was T4. The recommended canine dosage of levothyroxine may be too high for the treatment of hypothyroidism in American black bears; 0.011 mg/kg p.o. b.i.d. may be a more appropriate dosage. PMID:15193079

  9. Plasma levels of ursodeoxycholic acid in black bears, Ursus americanus: seasonal changes.

    PubMed

    Solá, Susana; Garshelis, David L; Amaral, Joana D; Noyce, Karen V; Coy, Pam L; Steer, Clifford J; Iaizzo, Paul A; Rodrigues, Cecília M P

    2006-06-01

    To date, no other studies have examined the seasonal changes in circulating levels of various bile acids in the plasma of wild North American black bears, Ursus americanus. Using gas chromatography, bile acid concentrations were measured in plasma samples obtained during either early or late hibernation, and during summer active periods. Thus, specific compositional changes from individual animals were examined through a given year. Total bile acid concentrations in the plasma of these normal animals were found to range between 0.2 and 3.1 micromol/L (0.9 +/- 0.2 micromol/L, mean +/- SEM). Cholic, ursodeoxycholic and chenodeoxycholic acids were the major bile acid species identified. Ursodeoxycholic acid represented 28.0 +/- 2.6% of the total bile acid pool. Deoxycholic and lithocholic acids were found only in small amounts. In addition, total bile acid concentrations were lower in plasma samples obtained during hibernation compared with those obtained during summer active periods (0.6 +/- 0.1 and 1.2 +/- 0.4 micromol/L, respectively; p < 0.05). However, the relative proportion of ursodeoxycholic acid, was significantly greater in winter than in summer (31.5 +/- 3.2% and 22.2 +/- 4.5%, p < 0.05). Finally, taurine-conjugated bile acids were the predominant species in bear plasma, accounting for >67% of the total bile acids. These data demonstrate that ursodeoxycholic acid is a major bile acid in black bear plasma, mostly conjugated with taurine. Further, the finding of seasonal variation in plasma bile acid composition provides evidence to support the possible role that ursodeoxycholic acid may play in cellular protection in hibernating black bears. PMID:16571381

  10. Fatal Caprine arthritis encephalitis virus-like infection in 4 Rocky Mountain goats (Oreamnos americanus).

    PubMed

    Patton, Kristin M; Bildfell, Robert J; Anderson, Mark L; Cebra, Christopher K; Valentine, Beth A

    2012-03-01

    Over a 3.5-year period, 4 Rocky Mountain goats (Oreamnos americanus), housed at a single facility, developed clinical disease attributed to infection by Caprine arthritis encephalitis virus (CAEV). Ages ranged from 1 to 10 years. Three of the goats, a 1-year-old female, a 2-year-old male, and a 5-year-old male, had been fed raw domestic goat milk from a single source that was later found to have CAEV on the premises. The fourth animal, a 10-year-old male, had not ingested domestic goat milk but had been housed with the other 3 Rocky Mountain goats. All 4 animals had clinical signs of pneumonia prior to death. At necropsy, findings in lungs included marked diffuse interstitial pneumonia characterized histologically by severe lymphoplasmacytic infiltrates with massive alveolar proteinosis, interstitial fibrosis, and type II pneumocyte hyperplasia. One animal also developed left-sided hemiparesis, and locally extensive lymphoplasmacytic myeloencephalitis was present in the cranial cervical spinal cord. Two animals had joint effusions, as well as severe lymphoplasmacytic and ulcerative synovitis. Immunohistochemical staining of fixed sections of lung tissue from all 4 goats, as well as spinal cord in 1 affected animal, and synovium from 2 affected animals were positive for CAEV antigen. Serology testing for anti-CAEV antibodies was positive in the 2 goats tested. The cases suggest that Rocky Mountain goats are susceptible to naturally occurring CAEV infection, that CAEV from domestic goats can be transmitted to this species through infected milk and by horizontal transmission, and that viral infection can result in clinically severe multisystemic disease.

  11. Multilocus heterozygosity, parental relatedness and individual fitness components in a wild mountain goat, Oreamnos americanus population.

    PubMed

    Mainguy, Julien; Côté, Steeve D; Coltman, David W

    2009-05-01

    Matings between relatives lead to a decrease in offspring genetic diversity which can reduce fitness, a phenomenon known as inbreeding depression. Because alpine ungulates generally live in small structured populations and often exhibit a polygynous mating system, they are susceptible to inbreeding. Here, we used marker-based measures of pairwise genetic relatedness and inbreeding to investigate the fitness consequences of matings between relatives in a long-term study population of mountain goats (Oreamnos americanus) at Caw Ridge, Alberta, Canada. We first assessed whether individuals avoided mating with kin by comparing actual and random mating pairs according to their estimated genetic relatedness, which was derived from 25 unlinked polymorphic microsatellite markers and reflected pedigree relatedness. We then examined whether individual multilocus heterozygosity H, used as a measure of inbreeding, was predicted by parental relatedness and associated with yearling survival and the annual probability of giving birth to a kid in adult females. Breeding pairs identified by genetic parentage analyses of offspring that survived to 1 year of age were less genetically related than expected under random matings. Parental relatedness was negatively correlated with offspring H, and more heterozygous yearlings had higher survival to 2 years of age. The probability of giving birth was not affected by H in adult females. Because kids that survived to yearling age were mainly produced by less genetically related parents, our results suggest that some individuals experienced inbreeding depression in early life. Future research will be required to quantify the levels of gene flow between different herds, and evaluate their effects on population genetic diversity and dynamics.

  12. Behavioral Thermoregulation and Trade-Offs in Juvenile Lobster Homarus americanus.

    PubMed

    Nielsen, Travis V; McGaw, Iain J

    2016-02-01

    Water temperature influences the behavior and distribution patterns of both larval and adult American lobster Homarus americanus. However, very little is known about the responses of juvenile lobsters. The juvenile life stage is a critical period; high levels of mortality, combined with specific behavioral responses, can disconnect larval settlement from patterns of abundance of adults. We assessed behavioral thermoregulation in juvenile lobsters, and determined how thermal preferences can be altered by the presence of shelter and food. Juvenile lobsters avoided temperatures higher than 20 °C and lower than 8 °C, and had a mean temperature preference of 16.2 ± 1 °C. This preference was unaffected by prior acclimation, origin (laboratory-raised or wild), or size. When the animals were subjected to a temperature change (5-20 °C), activity rates peaked at 15 °C, and remained stable thereafter. Activity rates did not change when a shelter was added. The addition of food resulted in an increase in activity associated with food handling. When juvenile lobsters were offered a choice between temperature, shelter, and food, they always chose the environment with a shelter, even when it was in a thermally unfavorable temperature. Juveniles also spent more time in a thermally unfavorable environment when food was present; however, acquisition of a shelter was prioritized over food. Although juveniles had a similar thermal preference to adults, they are more vulnerable to predation; the innate shelter-seeking behavior of juveniles overrode their thermal preference. While temperature is an important environmental factor affecting the physiology, distribution, and growth of aquatic ectotherms, our findings suggest that trade-off behaviors occur in order to maintain optimal fitness and survival of the individual. PMID:26896176

  13. Population trends and flight behavior of the American burying beetle, Nicrophorus americanus (Coleoptera: Silphidae), on Block Island, RI

    USGS Publications Warehouse

    Raithel, C.J.; Ginsberg, H.S.; Prospero, M.L.

    2006-01-01

    The endangered American burying beetle, Nicrophorus americanus, was monitored on Block Island, RI, USA, from 1991-2003 using mark-recapture population estimates of adults collected in pitfall traps. Populations increased through time, especially after 1994 when a program was initiated that provided carrion for beetle production. Beetle captures increased with increasing temperature and dew point, and decreased with increasing wind speed. Short distance movement was not related to wind direction, while longer distance flights tended to be downwind. Although many individuals flew considerable distances along transects, most recaptures were in traps near the point of release. These behaviors probably have counterbalancing effects on population estimates.

  14. Species profiles: life histories and environmental requirements of coastal fishes and invertebrates (North Atlantic). American lobster. [Homarus americanus

    SciTech Connect

    MacKenzie, C.; Moring, J.R.

    1985-04-01

    This species profile is a literature summary of the taxonomy, morphology, distribution, life history, and environmental requirements of the American lobster (Homarus americanus). These species profiles are designed to assist in the preparation of environmental impact assessments. The American lobster is a valuable commercial shellfish. After spawning, lobsters undergo a series of molts; as adults they live in coastal and offshore waters. Lobsters are captured in baited traps and incidentally in trawls. About 5 to 6 million pounds were captured commercially in 1983, a downward trend from 1971. Major environmental factors affecting reproduction, growth, and survival are water temperature, oxygen concentration, salinity, and substrate. 82 refs., 2 figs., 3 tabs.

  15. On the ecology of two sympatric flounders of the genus Paralichthys in the Bay of Coquimbo, Chile

    NASA Astrophysics Data System (ADS)

    Acuña, Enzo; Cid, Luis

    The ecology of two flounders belonging to the genus Paralichthys was studied in the Bay of Coquimbo (29°57'S), Chile. P. adspersus grows larger than P. microps. In both species, the females were significantly larger than the males. The species differed in their reproductive cycles as revealed by their gonadosomatic indices. This difference was also reflected in larval abundance and spatial distribution of the species. No significant changes were found in the abundances of juveniles and adults in the Bay of Coquimbo either throughout the year or interannually. However, when the analysis included the proportion of specimens between the two species and sex by season, the differences were apparent in most cases. The results show that the environmental characteristics of the marine system which the Bay of Coquimbo is a part of provide good feeding, spawning and nursery grounds for these two species of flounder.

  16. De Novo Assembly of the Japanese Flounder (Paralichthys olivaceus) Spleen Transcriptome to Identify Putative Genes Involved in Immunity

    PubMed Central

    Huang, Lin; Li, Guiyang; Mo, Zhaolan; Xiao, Peng; Li, Jie; Huang, Jie

    2015-01-01

    Background Japanese flounder (Paralichthys olivaceus) is an economically important marine fish in Asia and has suffered from disease outbreaks caused by various pathogens, which requires more information for immune relevant genes on genome background. However, genomic and transcriptomic data for Japanese flounder remain scarce, which limits studies on the immune system of this species. In this study, we characterized the Japanese flounder spleen transcriptome using an Illumina paired-end sequencing platform to identify putative genes involved in immunity. Methodology/Principal Findings A cDNA library from the spleen of P. olivaceus was constructed and randomly sequenced using an Illumina technique. The removal of low quality reads generated 12,196,968 trimmed reads, which assembled into 96,627 unigenes. A total of 21,391 unigenes (22.14%) were annotated in the NCBI Nr database, and only 1.1% of the BLASTx top-hits matched P. olivaceus protein sequences. Approximately 12,503 (58.45%) unigenes were categorized into three Gene Ontology groups, 19,547 (91.38%) were classified into 26 Cluster of Orthologous Groups, and 10,649 (49.78%) were assigned to six Kyoto Encyclopedia of Genes and Genomes pathways. Furthermore, 40,928 putative simple sequence repeats and 47, 362 putative single nucleotide polymorphisms were identified. Importantly, we identified 1,563 putative immune-associated unigenes that mapped to 15 immune signaling pathways. Conclusions/Significance The P. olivaceus transciptome data provides a rich source to discover and identify new genes, and the immune-relevant sequences identified here will facilitate our understanding of the mechanisms involved in the immune response. Furthermore, the plentiful potential SSRs and SNPs found in this study are important resources with respect to future development of a linkage map or marker assisted breeding programs for the flounder. PMID:25723398

  17. Molecular Response of Estuarine Fish to Hypoxia: A Comparative Study with Ruffe and Flounder from Field and Laboratory

    PubMed Central

    Tiedke, Jessica; Thiel, Ralf; Burmester, Thorsten

    2014-01-01

    On a global scale, the frequencies and magnitudes of hypoxic events in coastal and estuarine waters have increased dramatically over the past 20 years. Fish populations are suitable indicators for the assessment of the quality of aquatic ecosystems, as they are omnipresent and often comprise a variety of different lifestyles and adaption strategies. We have investigated on the molecular level the impact of hypoxia on two fish species typical of European estuaries. We monitored the expression of eleven putatively hypoxia-responsive genes by means of quantitative real-time RT-PCR in brains, gills and hearts of the ruffe (Gymnocephalus cernua) and the flounder (Platichthys flesus). We first investigated the effect of naturally occurring hypoxia in the Elbe estuary. In a second approach, expression changes in the response to hypoxia were monitored under controlled laboratory conditions. The genes that showed the strongest effect were two respiratory proteins, myoglobin and neuroglobin, as well as the apoptosis enzyme caspase 3. As previously observed in other fish, myoglobin, which was considered to be muscle-specific, was found in brain and gills as well. Comparison of field and laboratory studies showed that – with the exception of the heart of flounder – that mRNA levels of the selected genes were about the same, suggesting that laboratory conditions reflect natural conditions. Likewise, trends of gene expression changes under hypoxia were the same, although hypoxia response was more pronounced in the Elbe estuary. In general, the flounder displayed a stronger response to hypoxia than the ruffe, suggesting that the flounder is more susceptible to hypoxia. The most pronounced differences were found among tissues within a species, demonstrating that hypoxia response is largely tissue-specific. In summary, our data suggest that laboratory experiments essentially mimic field data, but additional environmental factors enhance hypoxia response in nature. PMID:24595439

  18. Characterization of Paralichthys olivaceus peroxisome proliferator-activated receptor-α gene as a master regulator of flounder lipid metabolism.

    PubMed

    Cho, Hyun Kook; Kong, Hee Jeong; Kim, Hye Young; Cheong, JaeHun

    2012-01-01

    Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that play key roles in lipid and energy homeostasis. Paralichthys olivaceus PPARα (PoPPARα) cDNA was isolated by initial reverse transcription-polymerase chain reaction (RT-PCR) using conserved region among fish species and rapid amplification of cDNA ends (RACE). The full-length of PoPPARα cDNA is 2040-bp long encoding a polypeptide with 505 amino acids and containing a DNA binding domain (C4-type zinc finger) and a ligand-binding domain. PoPPARα was detected from 1 day post-hatch and was highly expressed in the stomach, liver, and intestine of continuously fed flounder, approximately 16 cm in size. PoPPARα mRNA expression was down-regulated in the kidney, stomach, and liver of the 4.5-month-old flounder after a 30 day food-deprivation period. PoPPARα activates the PPAR response element (PPRE)-driven reporter, and treatment with Wy14643, a PPARα agonist, augmented PoPPARα-stimulated peroxisome proliferator response element activity in HINAE and HepG2 cells. PoPPARα activated the expression of fatty acid β-oxidation related genes such as carnitine palmitoyltransferase 1A, medium chain acyl-CoA dehydrogenase, and acyl-CoA oxidase 1 and inhibited the expression of sterol regulatory element binding protein and fatty acid synthase by competitively inhibiting LXR/RXR heterodimer formation. These results suggest that PoPPARα plays an important role in lipid metabolism of olive flounder and that it is functionally and evolutionarily conserved in olive flounder and mammals. PMID:22051321

  19. Mutations in exons of the CYP17-II gene affect sex steroid concentration in male Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Ma, Ruiqin; He, Feng; Wen, Haishen; Li, Jifang; Shi, Bao; Shi, Dan; Liu, Miao; Mu, Weijie; Zhang, Yuanqing; Hu, Jian; Han, Weiguo; Zhang, Jianan; Wang, Qingqing; Yuan, Yuren; Liu, Qun

    2012-03-01

    As a specific gene of fish, cytochrome P450c17-II ( CYP17-II) gene plays a key role in the growth, development an reproduction level of fish. In this study, the single-stranded conformational polymorphism (SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-II gene in a population of 75 male Japanese flounder ( Paralichthys olivaceus). Three single nucleotide polymorphisms (SNPs) were identified in CYP17-II gene of Japanese flounder. They were c.G594A (p.G188R), c.G939A and c.G1502A (p.G490D). SNP1 (c.G594A), located in exon 4 of CYP17-II gene, was significantly associated with gonadosomatic index (GSI). Individuals with genotype GG of SNP1 had significantly lower GSI ( P < 0.05) than those with genotype AA or AG. SNP2 (c.G939A) located at the CpG island of CYP17-II gene. The mutation changed the methylation of exon 6. Individuals with genotype AA of SNP2 had significantly lower serum testosterone (T) level and hepatosomatic index (HSI) compared to those with genotype GG. The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder. However, the SNP3 (c.G1502A) located in exon 9 did not affect the four measured reproductive traits. This study showed that CYP17-II gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.

  20. Radiocesium biokinetics in olive flounder inhabiting the Fukushima accident-affected Pacific coastal waters of eastern Japan.

    PubMed

    Tateda, Yutaka; Tsumune, Daisuke; Tsubono, Takaki; Aono, Tatsuo; Kanda, Jota; Ishimaru, Takashi

    2015-09-01

    Radiocesium ((134)Cs and (137)Cs) originating from the Fukushima Dai-ichi Nuclear Power Plant (1FNPP) has contaminated coastal waters and been subsequently transferred to the marine biota along the Pacific coastal region of eastern Japan. To clarify the mechanism of radiocesium biokinetics in olive flounder, a commercially valuable and piscivorous predator, the biokinetics of (137)Cs was simulated using a dynamic biological compartment model and then validated with the measured concentrations in available monitoring data. The (137)Cs concentrations in seawater of the Pacific coastal sites of eastern Japan, from Kesen-numa (170 km north from the 1FNPP) to Choshi (190 km south from the 1FNPP), were reconstructed by fitting the simulated levels to the observed concentrations. Simulated values were verified by measured radiocesium levels in sedentary organism such as macro-algae and mussels inhabiting each study site which had accumulated radiocesium in their ambient environment from the beginning of the accident. Using reconstructed (137)Cs concentrations in seawater, the (137)Cs levels in olive flounder and its main planktivorous prey fish, e.g. anchovy, sand lance, whitebait, etc., were simulated and compared with observed concentrations to clarify the biokinetics of radiocesium in these organisms. This assessment showed that the determining factor for the maximum radiocesium concentrations in fish in the plankton food chain is likely to be the initial radiocesium concentration which they were exposed to during the contamination stage. Furthermore, the simulated (137)Cs concentrations in gut contents of olive flounder were verified by measured (137)Cs concentrations in the stomach contents of this fish collected within 30 km from the 1FNPP. These results indicated that the decrease of (137)Cs levels in their prey organisms was the primary determining factor of radiocesium depuration, and the resultant ecological half-lives were 140-160 d in the olive flounder

  1. Comparison of growth characteristics between skeletal muscle satellite cell lines from diploid and triploid olive flounder Paralichthys olivaceus

    PubMed Central

    Wu, Zhi-hao; Tan, Xungang; Jiao, Shuang; Zhang, Pei-jun

    2016-01-01

    Objectives. According to myosatellite cell lines (MSCs) established in vitro from diploid and triploid flounder, we compared the characters of growth and differentiation of their MSCs. The results would be useful for learning the muscle development mechanism in teleosts. Materials and Methods. The skeletal muscle cells from the diploid and triploid olive flounder Paralichthys olivaceus were isolated and cultured in vitro, respectively, and the cells were characterized at the morphology and molecular level; meanwhile, the performance of these cells’ proliferation and differentiation were analyzed. Results. Two new skeletal muscle cell lines (POMSCS(2n) and POMSCS(3n)) from diploid and triploid flounder have been respectively subcultured for 67 times and 66 times. The cultured cells were mostly spindle-like mononuclear cells. They have normal flounder diploid karyotype (2n=48t) and triploid karyotype (3n=72t), respectively. Muscle satellite cell gene marker (pax7b) and myogenic cell protein marker (Desmin) were all expressed in cells of two cell lines. Both of the cells could differentiate into the large polynucleated muscle fibre cells, and immunofluorescence reactions of myosin heavy chain (MyHC) were positive. There were more cells of POMSCS(3n) to differentiate into the muscle fibre cells than that of POMSCS(2n). However, POMSCS(2n) cells proliferated more rapidly than those of POMSCS(3n) (P < 0.05). The significant fluorescent signals were observed in both POMSCS(2n) and POMSCS(3n) cells after transfected with pEGFP-N3 reporter plasmid. Conclusions. The two cell lines have been established and characterized as MSCs. We suppose that it might be the differentiation capacity, rather than the proliferation activity of MSCs to play a key role in the better growth of triploid ones than diploid. Both cell lines will become the ideal tools to learn the mechanism of fish MSCs proliferation, differentiation and regeneration during muscle development in the future. PMID

  2. Food and feeding habits of juvenile flounder Platichthys flesus (L.), abd turbot Scophthalmus maximus L. in the åland archipelago, northern Baltic Sea

    NASA Astrophysics Data System (ADS)

    Aarnio, Katri; Bonsdorff, Erik; Rosenback, Nina

    1996-12-01

    The food choice of juvenile flounder ( Platichthys flesus) and turbot ( Scophthalmus maximus) was studied in the northern Baltic Sea during the years 1988, 1989, 1994 and 1995. The diet included organisms from 30 species/taxa in flounder (n = 306) and 10 species/taxa in turbot (n = 41). Flounder ⩽ 45 mm mainly consumed meiofauna (dominating taxon: Harpacticoida, Copepoda) and larger fish (46-101 mm) consumed macrofauna (dominating taxa: Oligochaeta, Amphipoda and Chironomidae). In terms of biomass, macrofauna dominated for all sizes of flounders, and meiofauna was important only for the smallest fish. A strong seasonal variation could be detected in the diet. In spring, macrofauna dominated for all size classes of fish (only fish > 30 mm were caught in spring), while in summer and autumn meiofauna dominated the diets for fish ⩽ 45 mm in size. Juvenile turbot (22-88 mm) consumed macrofauna and small fish. Turbot ⩽ 30 mm consumed mainly amphipods, while > 30 mm turbot consumed mysid shrimps, amphipods and fish. The ontogenetic shift from meio- to macrofauna-sized prey in flounders occurs at a larger fish size in the northern Baltic Sea than reported in other areas, possibly depending on the increased relative importance of meiofauna in the northern Baltic. The seasonal variation in the diet could be due to seasonally changing abundances in the zoobenthos, or for the small fish (1-group, spring), to switching from meio- to macrofauna in order to optimize their energy gain. The 0-group flounders consumed meiofauna for a long period, possibly due to a learning-process or simply due to easy availability of meiofauna. Turbot has a much larger mouth gap than flounders, thus allowing them to consume macrofauna from the beginning of their benthic life.

  3. Possible causes and consequences of variation in age and size at metamorphosis in flatfishes (pleuronectiformes): An analysis at the individual, population, and species levels

    NASA Astrophysics Data System (ADS)

    Chambers, R. C.; Leggett, W. C.

    Observations on metamorphic ages and sizes in flatfishes are synthesized within a hierarchical framework which ascends from individual to population to species levels. Examples of observations and inferences are given for each level with emphasis on variation among individuals within populations. At the species level, size at metamorphosis ranges from ≈4 to 120 mm and is log-normally distributed. Species-level estimates of ages at metamorphosis from field-collected flatfish are of low resolution if they are derived as the difference between approximate spawning and inferred settling dates. Temperature is the only environmental factor whose effects on metamorphic traits have been evaluated at the population level. Age at metamorphosis increases by more than twofold and metamorphic size is expected to increase slightly as temperature decreases across the viable range. At the individual level, data are available for starry flounder, Platichthys stellatus, and winter flounder, Pseudopleuronectes americanus. Bivariate distributions of metamorphic ages and sizes for these flounders, schematized as metamorphic envelopes, have different locations and orientations, but similar shapes. Metamorphic age has greater within-population variation and is less influenced by parentage than is metamorphic length. Furthermore, maternal contributions to metamorphic traits exceed paternal ones. Effects of metamorphic trait variation on size distributions of young-of-the-year juveniles were evaluated by simulations. In a benign environment lacking mortality bias, the effect of metamorphic age on juvenile size variation was ≈sixfold greater than the effect of metamorphic size. The impact of metamorphic age on juvenile size variation lessened with extension of the spawning interval and with increased duration of juvenile life prior to capture.

  4. Early development of circadian rhythmicity in the suprachiamatic nuclei and pineal gland of teleost, flounder (Paralichthys olivaeus), embryos.

    PubMed

    Mogi, Makoto; Uji, Susumu; Yokoi, Hayato; Suzuki, Tohru

    2015-08-01

    Circadian rhythms enable organisms to coordinate multiple physiological processes and behaviors with the earth's rotation. In mammals, the suprachiasmatic nuclei (SCN), the sole master circadian pacemaker, has entrainment mechanisms that set the circadian rhythm to a 24-h cycle with photic signals from retina. In contrast, the zebrafish SCN is not a circadian pacemaker, instead the pineal gland (PG) houses the major circadian oscillator. The SCN of flounder larvae, unlike that of zebrafish, however, expresses per2 with a rhythmicity of daytime/ON and nighttime/OFF. Here, we examined whether the rhythm of per2 expression in the flounder SCN represents the molecular clock. We also examined early development of the circadian rhythmicity in the SCN and PG. Our three major findings were as follows. First, rhythmic per2 expression in the SCN was maintained under 24 h dark (DD) conditions, indicating that a molecular clock exists in the flounder SCN. Second, onset of circadian rhythmicity in the SCN preceded that in the PG. Third, both 24 h light (LL) and DD conditions deeply affected the development of circadian rhythmicity in the SCN and PG. This is the first report dealing with the early development of circadian rhythmicity in the SCN in fish.

  5. Bacterial enteritis and the development of the larval digestive tract in olive flounder, Paralichthys olivaceus (Temminck & Schlegel).

    PubMed

    Kim, D-H; Han, H-J; Kim, S-M; Lee, D-C; Park, S-I

    2004-09-01

    Three bacterial isolates obtained from diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization and 16S rRNA gene sequencing studies. Bacterial enteritis was reproduced in 16 and 22 days post-hatch (dph) larvae by administering brine shrimp nauplii, Artemia salina, dosed with the environmental isolates and reference strains of V. ichthyoenteri. To investigate the effect of the disease on development of the stomach, a pepsin activity assay and reverse transcription-polymerase chain reaction (RT-PCR) analysis of the expression of the pepsinogen gene were performed. Expression of olive flounder pepsinogen was detected from 30-dph larvae and the increased level of pepsin activity coincided with reduced susceptibility to the disease. Growth rates of V. ichthyoenteri, V. anguillarum and Edwardsiella tarda were tested in artificial stomach conditions using HCl and porcine pepsin. All the strains of V. ichthyoenteri were inhibited by low pH conditions which corresponded with an increase in pepsin levels. This suggests that differentiation of the stomach in olive flounder larvae and juveniles, an essential physiological development, also provides the host with a non-immunological defence mechanism. PMID:15357708

  6. The effects of ambient pH on nitrogen excretion in early life stages of the American toad (Bufo americanus).

    PubMed

    Tattersall, G J; Wright, P A

    1996-04-01

    Acidification of breeding ponds has been identified as a potential threat to the survival and health of North American amphibian populations. The effects of acid exposure on ion and acid-balance are well known, but there is little information on how environmental water pH influences nitrogen balance in amphibians. The aim of this study was to determine the effects of moderately acidic water (pH 6.0) on nitrogen excretion in early life stages of the toad, Bufo americanus. Acid exposure (pH 6.0, 54 h) resulted in a 20-80% increase in ammonia-N excretion rates in embryos and early, middle and late tadpoles stages, whereas there was no significant effect on urea-N excretion. Tissue ammonia concentrations were significantly higher (+33%) in the embryos and 35-65% lower in the three groups of tadpoles exposed to water of pH 6.0 compared to control animals (pH 8.5). In embryos, ammonia excretion accounted for greater than 90% of total nitrogen excretion (ammonia-N + urea-N), but by the late tadpole stage this value had decreased to approximately 65%. These findings indicate that exposure of embryonic and larval B. americanus to moderately acidic water disrupts nitrogen balance by increasing nitrogen loss as ammonia, with no compensatory decrease in urea excretion.

  7. Manganese concentration in lobster (Homarus americanus) gills as an index of exposure to reducing conditions in western Long Island Sound

    USGS Publications Warehouse

    Draxler, Andrew F.J.; Sherrell, Robert M.; Wieczorek, Dan; Lavigne, Michele G.; Paulson, A.J.

    2005-01-01

    We examined the accumulation of manganese (Mn) in gill tissues of chemically nai??ve lobsters held in situ at six sites in Long Island Sound (LIS) for up to six weeks to evaluate the possible contribution of eutrophication-driven habitat quality factors to the 1999 mass mortality of American lobsters (Homarus americanus). These western LIS lobster habitats experience seasonal hypoxia, which results in redox-mobilized Mn being transferred to and deposited on the tissues of the lobsters. Manganese accumulated in gill tissue of lobsters throughout the study, but rates were highest at western and southern LIS sites, ranging from 3.4-0.8 ??g/g/d (???16 ??g/g initial). The Baden-Eriksson observation that Mn accumulation in Norway lobsters (Nephrops norvegicus) is associated with ecosystem hypoxia is confirmed and extended to H. americanus. It seems likely that, after accounting for molting frequency, certain critical values may be applied to other lobster habitats of the NE US shelf. If a high proportion of lobsters in autumn have gill Mn concentrations exceeding 30 ??g/g, then the habitats are likely experiencing some reduced oxygen levels. Manganese concentrations above 100 ??g/g suggest exposure to conditions with the potential for lobster mortality should the temperatures of bottom waters become elevated, and gill concentrations above some higher level (perhaps 300 ??g/g) indicate the most severe habitat conditions with a strong potential for hypoxia stress.

  8. Metabolism, food consumption and growth of plaice ( Pleuronectes platessa) and flounder ( Platichthys flesus) in relation to fish size and temperature

    NASA Astrophysics Data System (ADS)

    Fonds, M.; Cronie, R.; Vethaak, A. D.; Van Der Puyl, P.

    Daily rates of oxygen consumption, food consumption and growth of plaice ( Pleuronectes platessa) and flounder ( Platichthys flesus) have been measured in the laboratory at various constant temperatures. Oxygen consumption was related to body weight of the fish as a power function, with a weight exponent of between 0.71 and 0.85. No significant effects of temperature or feeding on this exponent were found. Flounder showed a significantly higher metabolic rate and a higher temperature coefficient for metabolism than plaice. Maximum daily rates of food consumption and the weight increment of fish fed with excess rations of fresh mussel meat could also be related to fish weights by means of power functions. For plaice these exponents decreased from about 0.9 at low temperatures (2-6 C°) to about 0.7 at high temperatures (18-22°C). Such a temperature effect on the weight exponent indicates that small juvenile fish eat more and grow faster at higher temperatures than do large older fish, and that large fish do better at low temperatures. After scaling of daily food consumption and growth in proportion to metabolic weights of the fish (W 0.78), feeding and growth at different fish sizes and temperatures can be compared and temperature-growth rate models can be used for investigations of feeding in natural populations. Compared to plaice, young flounder ate more and grew faster at higher temperatures (> 14°C). This may partly explain the preference of flounder for the shallower parts of coastal areas and estuaries, where summer temperatures and food densities are higher. Energy budgets of young plaice and flounder fed with excess rations of mussel meat indicate that at least 29% of the food energy is used for metabolism while about 37% of the food energy is converted into growth. The net conversion efficiency was estimated at 0.45 for food and growth in units of ash-free dry weight, and at 0.53 for food and growth in energy units. Analysis of the energy budget showed

  9. Candidatus Liberibacter americanus induces significant reprogramming of the transcriptome of the susceptible citrus genotype

    PubMed Central

    2013-01-01

    Background Citrus huanglongbing (HLB) disease is caused by endogenous, phloem-restricted, Gram negative, uncultured bacteria named Candidatus Liberibacter africanus (CaLaf), Ca. L. asiaticus (CaLas), and Ca. L. americanus (CaLam), depending on the continent where the bacteria were first detected. The Asian citrus psyllid vector, Diaphorina citri, transmits CaLas and CaLam and both Liberibacter species are present in Brazil. Several studies of the transcriptional response of citrus plants manifesting HLB symptoms have been reported, but only for CaLas infection. This study evaluated the transcriptional reprogramming of a susceptible genotype of sweet orange challenged with CaLam, using a customized 385K microarray containing approximately 32,000 unigene transcripts. We analyzed global changes in gene expression of CaLam-infected leaves of sweet orange during the symptomatic stage of infection and compared the results with previously published microarray studies that used CaLas-infected plants. Twenty candidate genes were selected to validate the expression profiles in symptomatic and asymptomatic PCR-positive leaves infected with CaLas or CaLam. Results The microarray analysis identified 633 differentially expressed genes during the symptomatic stage of CaLam infection. Among them, 418 (66%) were upregulated and 215 (34%) were down regulated. Five hundred and fourteen genes (81%) were orthologs of genes from Arabidopsis thaliana. Gene set enrichment analysis (GSEA) revealed that several of the transcripts encoded transporters associated with the endomembrane system, especially zinc transport. Among the most biologically relevant gene transcripts in GSEA were those related to signaling, metabolism and/or stimulus to hormones, genes responding to stress and pathogenesis, biosynthesis of secondary metabolites, oxidative stress and transcription factors belonging to different families. Real time PCR of 20 candidate genes validated the expression pattern of some genes in

  10. Seasonal changes in spermatogenesis and testicular steroidogenesis in the male black bear Ursus americanus.

    PubMed

    Tsubota, T; Howell-Skalla, L; Nitta, H; Osawa, Y; Mason, J I; Meiers, P G; Nelson, R A; Bahr, J M

    1997-01-01

    American black bears, Ursus americanus, are seasonal breeders with a mating season in late spring to early summer. The objectives of this study were to determine whether there are seasonal changes in spermatogenesis and immunolocalization of testicular steroidogenic enzymes, and to correlate these changes with peripheral steroid concentrations. Three captive mature bears were maintained in open cages during the summer season and provided with chambers for denning during the winter. Testicular biopsies and blood samples were obtained from anaesthetized bears on 12 March, 15 June, 12 October and 15 January. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3 beta-hydroxysteroid dehydrogenase (3 beta HSD), porcine testicular 17 alpha-hydroxylase cytochrome P450 (P450c17) and human placental aromatase cytochrome P450 (P450arom). Spermatogenesis changed seasonally: spermatogonia and degenerating spermatocytes were observed in October; spermatogonia and primary spermatocytes were present in January; spermatogonia, spermatocytes and round spermatids were present in March; and spermatogonia through spermatozoa were present in June. P450scc and P450c17 were immunolocalized in spermatids and Leydig cells in June, whereas in October these enzymes were present only in Leydig cells. 3 beta HSD was localized in Leydig cells in June and October with more intense staining in June. Localization of P450arom changed seasonally: no immunostaining in October; positive immunostaining in Sertoli cells in January; more extensive immunostaining in Sertoli cells, peritubular-myoid cells and round spermatids in March; and strong immunostaining in Sertoli cells and round and elongating spermatids in June. Serum testosterone and oestradiol concentrations changed seasonally: testosterone and oestrogen were low in October and January, slightly higher in March, and high in

  11. Ocean acidification effects in the early life-stages of summer flounder, Paralichthys dentatus

    NASA Astrophysics Data System (ADS)

    Chambers, R. C.; Candelmo, A. C.; Habeck, E. A.; Poach, M. E.; Wieczorek, D.; Cooper, K. R.; Greenfield, C. E.; Phelan, B. A.

    2013-08-01

    The limited available evidence about effects of high CO2 and acidification of our oceans on fish suggests that effects will differ across fish species, be subtle, and interact with other stressors. An experimental framework was implemented that includes the use of (1) multiple marine fish species of relevance to the northeastern USA that differ in their ecologies including spawning season and habitat; (2) a wide yet realistic range of environmental conditions (i.e., concurrent manipulation of CO2 levels and water temperatures), and (3) a diverse set of response variables related to fish sensitivity to elevated CO2 levels, water temperatures, and their interactions. This report is on an array of early life-history responses of summer flounder (Paralichthys dentatus), an ecologically and economically important flatfish of this region, to a wide range of pH and CO2 levels. Survival of summer flounder embryos was reduced by 50% below local ambient conditions (7.8 pH, 775 ppm pCO2) when maintained at the intermediate conditions (7.4 pH, 1860 ppm pCO2), and by 75% below local ambient when maintained at the most acidic conditions tested (7.1 pH, 4715 ppm pCO2). This pattern of reduced survival of embryos at higher CO2 levels was consistent among three females used as sources of embryos. Sizes and shapes of larvae were altered by elevated CO2 levels with longer larvae in more acidic waters. This pattern of longer larvae was evident at hatching (although longer hatchlings had less energy reserves) to midway through the larval period. Larvae from the most acidic conditions initiated metamorphosis at earlier ages and smaller sizes than those from more moderate and ambient conditions. Tissue damage was evident in older larvae (age 14 to 28 d post-hatching) from both elevated CO2 levels. Damage included liver sinusoid dilation, focal hyperplasia on the epithelium, separation of the trunk muscle bundles, and dilation of the liver sinusoids and central veins. Cranial

  12. Sequential developmental acquisition of cotransmitters in identified sensory neurons of the stomatogastric nervous system of the lobsters, Homarus americanus and Homarus gammarus.

    PubMed

    Kilman, V; Fénelon, V S; Richards, K S; Thirumalai, V; Meyrand, P; Marder, E

    1999-06-01

    We studied the developmental acquisition of three of the cotransmitters found in the gastropyloric receptor (GPR) neurons of the stomatogastric nervous systems of the lobsters Homarus americanus and Homarus gammarus. By using wholemount immunocytochemistry and confocal microscopy, we examined the distribution of serotonin-like, allatostatin-like, and FLRF(NH2)-like immunoreactivities within the stomatogastric nervous system of embryonic, larval, juvenile, and adult animals. The GPR neurons are peripheral sensory neurons that send proprioceptive information to the stomatogastric and commissural ganglia. In H. americanus, GPR neurons of the adult contain serotonin-like, allatostatin-like, and Phe-Leu-Arg-Phe-amide (FLRF(NH2))-like immunoreactivities. In the stomatogastric ganglion (STG) of the adult H. americanus and H. gammarus, all of the serotonin-like and allatostatin-like immunoreactivity colocalizes in neuropil processes that are derived exclusively from ramifications of the GPR neurons. In both species, FLRF(NH2)-like immunoreactivity was detected in the STG neuropil by 50% of embryonic development (E50). Allatostatin-like immunoreactivity was visible first in the STG at approximately E70-E80. In contrast, serotonin staining was not clearly visible until larval stage I (LI) in H. gammarus and until LII or LIII in H. americanus. These data indicate that there is a sequential acquisition of the cotransmitters of the GPR neurons. PMID:10340509

  13. ISOLATION AND GENETIC CHARACTERIZATION OF TOXOPLASMA GONDII FROM RACCOONS (PROCYON LOTOR), CATS (FELIS DOMESTICUS), STRIPED SKUNK (MEPHITIS MEPHITIS), BLACK BEAR (URSUS AMERICANUS), AND COUGAR (PUMA CONCOLOR) FROM CANADA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats ( Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus ) from Kuujjuaq, northern Quebec, Canada. Geno...

  14. Population ecology and habitat preferences of juvenile flounder Platichthys flesus (Actinopterygii: Pleuronectidae) in a temperate estuary

    NASA Astrophysics Data System (ADS)

    Souza, Allan T.; Dias, Ester; Nogueira, Ana; Campos, Joana; Marques, João C.; Martins, Irene

    2013-05-01

    The European flounder Platichthys flesus is a widely distributed epibenthic species and an important component of demersal fish assemblages in the European Atlantic coastal waters. In Portuguese estuaries, this species reaches high densities, especially in Minho estuary (NW Iberian Peninsula, Europe), potentially playing an important role in the system's ecology. In this context, the population structure, production and the habitat use of juvenile P. flesus were investigated. Sampling took place monthly, from February 2009 until July 2010 along the entire estuarine gradient (5 sampling stations distributed in the first 29 km from the river mouth, with S1 located near the river mouth, S2 inside a salt marsh, S3 in a salinity transition zone, while S4 and S5 were located in the upper estuary). Flounder's density varied significantly among sampling stations and seasons (two-way PERMANOVA: p < 0.001), with the majority of the individuals being found during the spring (30.1%) and in S3 and S4 (72.6%). Males and females presented an even distribution, with a higher proportion of males observed during summer. Fish length also differed among sampling stations and seasons (two-way PERMANOVA: p < 0.001), with larger fishes being found in S1 during the autumn (168.50 ± 59.50 mm) and the smallest in S4 during the spring (33.80 ± 3.12 mm). Size classes associated differently with environmental variables, with larger juveniles being more abundant in the downstream areas of the estuary, whereas smaller juveniles were related to higher water temperatures, suggesting a habitat segregation of P. flesus of different sizes. The fish condition of P. flesus in Minho estuary was higher than in other systems, probably due to the dominance of juveniles on the population. Also, the densities found in this estuary were up to 32 times higher than in other locations, suggesting that Minho estuary is an important nursery area for the species. The estimated secondary production of P. flesus

  15. Application of a food chain model to polychlorinated biphenyl contamination of the lobster and winter flounder food chains in New Bedford Harbor

    SciTech Connect

    Connolly, J.P. Manhattan Coll., Riverdale, NY )

    1991-04-01

    As part of a Remedial Investigation/Feasibility Study for the New Bedford Harbor Superfund site a model of polychlorinated biphenyls (PCBs) in the lobster and winter flounder food chains was developed. This model successfully reproduces tri-, tetra-, penta-, and hexachlorobiphenyl concentrations observed at all levels of the food chain and across the 2 order of magnitude concentration gradient in the system. The model indicated that PCB concentrations in the flounder and, to a lesser extent, in the lobster are derived from the sediment. Dietary uptake exceeds uptake across the gill for all four homologues and becomes the dominant route at the higher chlorinated homologues. The assimilation efficiency of ingested PCB apparently declines from relatively high values for tri-chlorobiphenyl to relatively low values for hexachlorobiphenyl. Differences in observed lobster and flounder PCB concentrations appear to be due to differences in the importance of the benthic component of the food chains of these animals and differences in whole body lipid content.

  16. Cloning and characterization of apoptosis-associated speck-like protein containing a CARD domain (ASC) gene from Japanese flounder Paralichthys olivaceus.

    PubMed

    Li, Shuo; Chen, Xiaoli; Peng, Weijiao; Hao, Gaixiang; Geng, Xuyun; Zhan, Wenbin; Sun, Jinsheng

    2016-07-01

    Apoptosis-associated speck-like protein containing a CARD domain (ASC) is a critical adaptor molecule in multiple inflammasome protein complexes that mediate inflammation and host defense. However, few studies have been performed in lower vertebrates such as in teleost. Here we identified and characterized a novel ASC gene (namely PoASC) from Japanese flounder Paralichthys olivaceus. The complete cDNA sequence of PoASC contains a 22 bp 5'-untranslated sequence, a 612 bp open reading frame, and a 438 bp 3'-untranslated sequence. The deduced PoASC protein is comprised of 203 amino acids with a conserved N-terminal PYD domain and a C-terminal CARD domain and shows 35-62% sequence identity with other vertebrate ASC proteins. PoASC mRNA transcripts was detected in various Japanese flounder tissues and is dominantly expressed in hepatopancreas. Oligomeric speck-like structures were observed when PoASC was exogenously expressed in Japanese flounder FG-9307 cells. Immune challenge experiments revealed that PoASC gene expression was significantly induced in the Japanese flounder head kidney macrophages and peripheral blood leukocytes by the canonical TLR ligands LPS, Poly(I:C) and zymosan stimulations. In addition, the induction of PoASC was also observed in Edwardsiella tarda challenged head kidney and gill tissues. Furthermore, we for the first time showed that extracellular ATP, an important signaling molecule in triggering innate immune response and activation of NLR inflammasome, significantly up-regulates PoASC expression in the Japanese flounder head kidney macrophages in a dose-dependent manner. Together, these findings addressed the involvement of PoASC in TLR and extracellular ATP-mediated innate immune signaling in the Japanese flounders.

  17. Development and Validation of Single Nucleotide Polymorphism (SNP) Markers from an Expressed Sequence Tag (EST) Database in Olive Flounder (Paralichthys olivaceus).

    PubMed

    Kim, Jung Eun; Lee, Young Mee; Lee, Jeong-Ho; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Kim, Kyung-Kil

    2014-12-01

    To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

  18. Cloning and characterization of apoptosis-associated speck-like protein containing a CARD domain (ASC) gene from Japanese flounder Paralichthys olivaceus.

    PubMed

    Li, Shuo; Chen, Xiaoli; Peng, Weijiao; Hao, Gaixiang; Geng, Xuyun; Zhan, Wenbin; Sun, Jinsheng

    2016-07-01

    Apoptosis-associated speck-like protein containing a CARD domain (ASC) is a critical adaptor molecule in multiple inflammasome protein complexes that mediate inflammation and host defense. However, few studies have been performed in lower vertebrates such as in teleost. Here we identified and characterized a novel ASC gene (namely PoASC) from Japanese flounder Paralichthys olivaceus. The complete cDNA sequence of PoASC contains a 22 bp 5'-untranslated sequence, a 612 bp open reading frame, and a 438 bp 3'-untranslated sequence. The deduced PoASC protein is comprised of 203 amino acids with a conserved N-terminal PYD domain and a C-terminal CARD domain and shows 35-62% sequence identity with other vertebrate ASC proteins. PoASC mRNA transcripts was detected in various Japanese flounder tissues and is dominantly expressed in hepatopancreas. Oligomeric speck-like structures were observed when PoASC was exogenously expressed in Japanese flounder FG-9307 cells. Immune challenge experiments revealed that PoASC gene expression was significantly induced in the Japanese flounder head kidney macrophages and peripheral blood leukocytes by the canonical TLR ligands LPS, Poly(I:C) and zymosan stimulations. In addition, the induction of PoASC was also observed in Edwardsiella tarda challenged head kidney and gill tissues. Furthermore, we for the first time showed that extracellular ATP, an important signaling molecule in triggering innate immune response and activation of NLR inflammasome, significantly up-regulates PoASC expression in the Japanese flounder head kidney macrophages in a dose-dependent manner. Together, these findings addressed the involvement of PoASC in TLR and extracellular ATP-mediated innate immune signaling in the Japanese flounders. PMID:27103005

  19. Evolution history of duplicated smad3 genes in teleost: insights from Japanese flounder, Paralichthys olivaceus

    PubMed Central

    Du, Xinxin; Liu, Yuezhong; Liu, Jinxiang; Zhang, Quanqi

    2016-01-01

    Following the two rounds of whole-genome duplication (WGD) during deuterosome evolution, a third genome duplication occurred in the ray-fined fish lineage and is considered to be responsible for the teleost-specific lineage diversification and regulation mechanisms. As a receptor-regulated SMAD (R-SMAD), the function of SMAD3 was widely studied in mammals. However, limited information of its role or putative paralogs is available in ray-finned fishes. In this study, two SMAD3 paralogs were first identified in the transcriptome and genome of Japanese flounder (Paralichthys olivaceus). We also explored SMAD3 duplication in other selected species. Following identification, genomic structure, phylogenetic reconstruction, and synteny analyses performed by MrBayes and online bioinformatic tools confirmed that smad3a/3b most likely originated from the teleost-specific WGD. Additionally, selection pressure analysis and expression pattern of the two genes performed by PAML and quantitative real-time PCR (qRT-PCR) revealed evidence of subfunctionalization of the two SMAD3 paralogs in teleost. Our results indicate that two SMAD3 genes originate from teleost-specific WGD, remain transcriptionally active, and may have likely undergone subfunctionalization. This study provides novel insights to the evolution fates of smad3a/3b and draws attentions to future function analysis of SMAD3 gene family. PMID:27703851

  20. Cloning and expression analysis of three novel CC chemokine genes from Japanese flounder (Paralichthys olivaceus).

    PubMed

    Zou, Gang-gang; Nozaki, Reiko; Kondo, Hidehiro; Hirono, Ikuo

    2014-10-01

    Chemokines are small cytokines secreted by various cell types. They not only function in cell activation, differentiation and trafficking, but they also have influences on many biological processes. In this study, three novel CC chemokine genes Paol-SCYA105, 106 and 107 in Japanese flounder (Paralichthys olivaceus) were cloned and characterized. Paol-SCYA105 was mainly detected in gill, kidney and spleen, Paol-SCYA106 was detected in all tissues examined and Paol-SCYA107 was mainly detected in the spleen and kidney. Paol-SCYA105 and Paol-SCYA106 gene expressions peaked in kidney at day 3 after viral hemorrhagic septicemia virus infection and decreased at day 6, but Paol-SCYA106 still remained at a high level at day 6. Paol-SCYA107 gene expression was significantly up-regulated in kidney at day 6 after viral hemorrhagic septicemia virus infection. In response to infection by Gram-negative Edwardsiella tarda and Gram-positive Streptococcus iniae in kidney, only Paol-SCYA106 gene expression significantly increased. Together, these results indicate that these three novel CC chemokines are involved in the immune response against pathogen infections.

  1. Lectin histochemistry of Kudoa septempunctata genotype ST3-infected muscle of olive flounder (Paralichthys olivaceus)

    PubMed Central

    Kang, Jaeyoun; Park, Changnam; Jang, Yeounghwan; Ahn, Meejung; Shin, Taekyun

    2016-01-01

    The localization of carbohydrate terminals in Kudoa septempunctata ST3-infected muscle of olive flounder (Paralichthys olivaceus) was investigated using lectin histochemistry to determine the types of carbohydrate sugar residues expressed in Kudoa spores. Twenty-one lectins were examined, i.e., N-acetylglucosamine (s-WGA, WGA, DSL-II, DSL, LEL, STL), mannose (Con A, LCA, PSA), galactose/N-acetylgalactosamine (RCA12, BSL-I, VVA, DBA, SBA, SJA, Jacalin, PNA, ECL), complex type N-glycans (PHA-E and PHA-L), and fucose (UEA-I). Spores encased by a plasmodial membrane were labeled for the majority of these lectins, with the exception of LCA, PSA, PNA, and PHA-L. Four lectins (RCA 120, BSL-I, DBA, and SJA) belonging to the galactose/N-acetylgalactosamine group, only labeled spores, but not the plasmodial membrane. This is the first confirmation that various sugar residues are present in spores and plasmodial membranes of K. septempunctata ST3. PMID:27169676

  2. Distribution of mucous cells on the body surface of Japanese flounder Paralichthys olivaceus.

    PubMed

    Yamamoto, T; Kawai, K; Oshima, S

    2011-03-01

    The distribution of mucous cells was examined in the skin on the ocular and blind sides of Japanese flounder Paralichthys olivaceus. Observations were performed on both body sides at the following regions: cheek, lower jaw (blind side), gill cover (ocular side), dorsal side, lateral line, belly and caudal peduncle. The mucous cells observed were elliptic and positively stained for periodic acid Schiff reaction and Mayer's mucicarmine and showed a higher density and larger size on the ocular side compared to the blind side. Low densities of mucous cells were observed on the lower jaw compared with other regions of the body. The depth of the crack located between scales was deeper on the ocular side than the blind side, which might reflect total epidermis area and total number of mucous cells. Bacterial infection elucidated some information on the effect on the density and size of mucous cells, where the density and size decreased slightly after infection. Only the lower jaw, however, showed an increased number of mucous cells. The results show that the potential of skin to secrete mucus is higher on the ocular than on the blind side and bacterial infection decreases mucous secretion. PMID:21366577

  3. Lectin histochemistry of Kudoa septempunctata genotype ST3-infected muscle of olive flounder (Paralichthys olivaceus).

    PubMed

    Kang, Jaeyoun; Park, Changnam; Jang, Yeounghwan; Ahn, Meejung; Shin, Taekyun

    2016-01-01

    The localization of carbohydrate terminals in Kudoa septempunctata ST3-infected muscle of olive flounder (Paralichthys olivaceus) was investigated using lectin histochemistry to determine the types of carbohydrate sugar residues expressed in Kudoa spores. Twenty-one lectins were examined, i.e., N-acetylglucosamine (s-WGA, WGA, DSL-II, DSL, LEL, STL), mannose (Con A, LCA, PSA), galactose/N-acetylgalactosamine (RCA12, BSL-I, VVA, DBA, SBA, SJA, Jacalin, PNA, ECL), complex type N-glycans (PHA-E and PHA-L), and fucose (UEA-I). Spores encased by a plasmodial membrane were labeled for the majority of these lectins, with the exception of LCA, PSA, PNA, and PHA-L. Four lectins (RCA 120, BSL-I, DBA, and SJA) belonging to the galactose/N-acetylgalactosamine group, only labeled spores, but not the plasmodial membrane. This is the first confirmation that various sugar residues are present in spores and plasmodial membranes of K. septempunctata ST3. PMID:27169676

  4. Establishment and characterization of a testicular Sertoli cell line from olive flounder Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Peng, Limin; Zheng, Yuan; You, Feng; Wu, Zhihao; Zou, Yuxia; Zhang, Peijun

    2016-09-01

    The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical "S" shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.

  5. Involvement of melanin-concentrating hormone 2 in background color adaptation of barfin flounder Verasper moseri.

    PubMed

    Mizusawa, Kanta; Kawashima, Yusuke; Sunuma, Toshikazu; Hamamoto, Akie; Kobayashi, Yuki; Kodera, Yoshio; Saito, Yumiko; Takahashi, Akiyoshi

    2015-04-01

    In teleosts, melanin-concentrating hormone (MCH) plays a key role in skin color changes. MCH is released into general circulation from the neurohypophysis, which causes pigment aggregation in the skin chromatophores. Recently, a novel MCH (MCH2) precursor gene, which is orthologous to the mammalian MCH precursor gene, has been identified in some teleosts using genomic data mining. The physiological function of MCH2 remains unclear. In the present study, we cloned the cDNA for MCH2 from barfin flounder, Verasper moseri. The putative prepro-MCH2 contains 25 amino acids of MCH2 peptide region. Liquid chromatography-electrospray ionization mass spectrometry with a high resolution mass analyzer were used for confirming the amino acid sequences of MCH1 and MCH2 peptides from the pituitary extract. In vitro synthesized MCH1 and MCH2 induced pigment aggregation in a dose-dependent manner. A mammalian cell-based assay indicated that both MCH1 and MCH2 functionally interacted with both the MCH receptor types 1 and 2. Mch1 and mch2 are exclusively expressed in the brain and pituitary. The levels of brain mch2 transcript were three times higher in the fish that were chronically acclimated to a white background than those acclimated to a black background. These results suggest that in V. moseri, MCH1 and MCH2 are involved in the response to changes in background colors, during the process of chromatophore control.

  6. Polluted harbor sediment and the annual reproductive cycle of the female flounder, Platichthys fiesus (L.)

    SciTech Connect

    Janssen, P.A.H.; Lambert, J.G.D.; Goos, H.J.T.; Wezel, A.P. van; Opperhuizen, A.

    1995-12-31

    Compounds such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs),and pesticides are metabolized by enzyme systems, which are also involved in steroid metabolism. Disturbances of reproduction may therefore occur through the interference of these compounds with the endocrine system. Several aspects of reproduction were studied in the flounder, Platichthys fiesus (L.), an euryhaline flatfish which inhabits coastal waters and is therefore a suitable biomonitor for the effects of chemical pollutants. Fish were kept during three years in mesocosm systems of which the first provided a control, while the second one contained polluted sediment, derived from the Rotterdam harbor. In November, all ovaries from both mesocosms contained vitellogenic oocytes. In May, all the control fish were previtellogenic, while the ovaries of fish from the polluted mesocosm contained, besides previtellogenic oocytes, a large number of vitellogenic oocytes, indicating that an estrogenic induction had occurred. The in vitro tissue incubations with androstenedione as precursor revealed that the ovarian capacity to synthesize testosterone (T), estrone (E{sub 2}) and 17{beta}-estradiol (E{sub 2}) didn`t differ between both mesocosms. In May, however, the levels of T and E{sub 2} as well as the level of the yolk-precursor vitellogenin were significantly higher in the polluted mesocosm. The conclusion from this study was that polluted harbor sediment contains compounds that effect normal reproductive development, i.e. the induction of premature vitellogenesis.

  7. Generation and characterization of monoclonal antibodies against the flounder Paralichthys olivaceus rhabdovirus.

    PubMed

    Zhou, Guang-Zhou; Gui, Lang; Li, Zheng-Qiu; Yuan, Xiu-Ping; Zhang, Qi-Ya

    2008-03-01

    Two MAbs (3C7 and 3C9) against flounder Paralichthys olivaceus rhabdovirus (PORV) were generated with hybridoma cell fusion technology and characterized by an indirect enzyme-linked immunosorbent assay, isotype test, Western blot and immunodot analysis and immunofluorescence assay. Isotyping tests demonstrated that both of the two MAbs belonged to IgM subclass. Western blot analysis showed the MAbs reacted with 42, 30, and 22 kDa viral proteins, which were localized within the cytoplasm of PORV-infected grass carp ovary (GCO) cells analyzed by indirect immunofluorescences tests. The MAb 3C7 was also selected at random for detecting virus antigens in the inoculated grass carp tissues by immunohistochemistry assay. Flow cytometry tests showed that at the 36 h postinfection (0.25 PFU/cell), the 23% PORV-infected GCO cells could be distinguished from the uninfected cells with the MAb 3C7. Such MAbs could be useful for diagnosis and potential treatment of viral infection.

  8. Distribution of marine birnavirus in cultured olive flounder Paralichthys olivaceus in Korea.

    PubMed

    Jung, Sung-Ju; Kim, Seok-Ryel; Joung, Il-Yong; Kitamura, Shin-Ichi; Ceong, Hee-Taek; Oh, Myung-Joo

    2008-06-01

    Surveys of marine birnavirus (MABV) were undertaken in cultured olive flounder Paralichthys olivaceus from the south and west coastal areas and Jeju in Korea during the period January 1999 to April 2007. MABV was detected in all seasons from the fry, juveniles and adult fish from the areas examined. Evident cytopathic effects of the virus including rounding and cell lysis were observed in chinook salmon embryo (CHSE-214) and rainbow trout gonad (RTG-2) cells, but not in fathead minnow (FHM) and epithelial papilloma of carp (EPC) cells. Nucleotide sequences of the VP2/NS junction region of the Korean isolates showed 97.8% ~ 100% similarity, and they belonged to the same genogroup. Cross neutralization tests with serotype-specific rabbit antisera against MABV strains exhibited a close antigenic relationships between strains, and were distinct from infectious pancreatic necrosis virus (IPNV) strains. Coinfection of MABV with bacteria (Streptococcus iniae, Vibrio spp.) and viruses (nervous necrosis virus, lymphocystis disease virus, viral hemorrhagic septicemia virus) was observed. PMID:18604495

  9. Induction of apoptosis in a flounder gill cell line by lymphocystis disease virus infection.

    PubMed

    Hu, G-B; Cong, R-S; Fan, T-J; Mei, X-G

    2004-11-01

    Lymphocystis disease virus (LCDV), a large icosahedral DNA virus classified to the iridovirus family, is the causative agent of lymphocystis, a disease which occurs in marine and freshwater fish species and is characterized by formation of papilloma-like lesions on the surface of the skin. In vitro, LCDV infection causes flounder gill cells, an adherent cell line, to exhibit an obvious cytopathic effect (CPE). In order to test whether apoptosis is responsible for the observed CPE, cells infected with LCDV at a multiplicity of infection (m.o.i.) of 5 PFU per cell were examined at various time intervals for the appearance of apoptotic signs. Nuclear fragmentation, DNA laddering and caspase activation were observed in the infected cells at the time (i.e. 10 days post-infection) when an intensive CPE was observed. These findings demonstrate that LCDV is capable of inducing apoptosis in vitro, which is different from the result of LCDV infection in vivo, and consequently suggest an intricate LCDV-host interaction. PMID:15509260

  10. Effects of cooking on levels of PCBs in the fillets of winter flounder

    SciTech Connect

    Posten, T.M.; Durell, G.S.; Moya, J.; Garrahan, K.G.

    1994-12-31

    The consumption of contaminated fish has been shown to be a significant, pathway for human exposure. Risk assessors often assume that humans are exposed to the levels of contaminants in edible tissue of fish measured prior to preparation and cooking. This assumption may lead to overestimation or underestimation of risk since a particular cooking method may remove or transform toxic constituents in the flesh by thermal denaturation, vaporization, dissolution in aqueous tissue fluids or lipids that drip off the flesh, or extraction into cooling oil during deep fat frying. This paper presents and discusses the results of a study conducted to quantify the effect of preparation and cooking on PCB concentrations in the edible portion of winter flounder. The effects of broiling, pan frying, and deep frying in oil were tested on fillets from 21 fish. The change in total PCB concentrations and 18 specific PCB congeners was estimated using a mass-balance approach using the precooked PCB concentration on a wet-weight basis and the cooked concentration taking into account any change in fillet weight resulting from cooking. Deep frying in oil resulted in a 47% reduction in total PCB levels in fillet tissue, while pan frying and broiling did not result in a statistically significant difference in total PCB levels.

  11. Philometrids of the southern flounder Paralichthys lethostigma: a multidimensional approach to determine their diversity.

    PubMed

    de Buron, Isaure; France, Scott G; Connors, Vincent A; Roumillat, William A; Tsoi, Lam C

    2011-06-01

    Two species of philometrid nematode, Philometra overstreeti and Philometroides paralichthydis, infect the southern flounder, Paralichthys lethostigma. Individuals of P. overstreeti are located between the teeth and inside the bony part of the branchial arches of the fish. Individuals of P. paralichthydis are associated with the bones of the buccal cavity and among muscles that control the dorsal and anal fins. Sequencing of part of the cytochrome oxidase I gene revealed 4 distinct genetic clades, each corresponding exactly to the 4 respective locations of the parasites in the host, suggesting the need for taxonomic revision. We hypothesized that each clade represented a separate species and, because the worms are morphologically indistinguishable, compared population level parameters of the clades comprising each currently recognized species. For each currently recognized species, the presence of worms from 1 clade was negatively correlated with the presence of worms from the other. Results also indicated significant differences between the clades in prevalences relative to both biotic and abiotic factors. Results clearly indicated major differences in the ecology of the philometrids constituting each clade. Taken as a whole, molecular and ecological data support the contention that the 4 genetic clades are likely 4 distinct species. PMID:21506857

  12. Passive and active ion transport by the urinary bladder of a euryhaline flounder.

    PubMed

    Demarest, J R; Machen, T E

    1984-04-01

    The effects of voltage clamping on the flux ratios and unidirectional and net fluxes of Na and Cl were used to gain insight into the mechanisms of active and passive ion transport across urinary bladders isolated from seawater-(SW) and freshwater-acclimated (FW) flounder, Platichthys stellatus. Although the transepithelial conductance (Gt = 2.77 mS X cm-2) of FW bladders was much greater than that of SW bladders (Gt = 0.40 mS X cm-2), the current-voltage relationships of both SW and FW bladders were markedly nonlinear. Under short-circuit conditions there was a large difference in the serosal-to-mucosal Na flux (JNasm) between SW (0.10 mueq X cm-2 X h-1) and FW (1.71 mueq X cm-2 X h-1) bladders, but their mannitol permeabilities were identical. The results indicate that 1) the paracellular pathway of both SW and FW bladders is Cl selective and Cl movements through the shunt account for a maximum of 90% of Gt in SW bladders and 19% in FW bladders; 2) the larger Gt of FW bladders is due to greater conductance of the apical cell membrane; 3) the majority of the passive ion movement across these epithelia proceeds through nonconductive, presumably transcellular, pathways; and 4) active transport of Na and Cl occurs by neutral coupling to each other and to other unidentified ions.

  13. Serologic Survey of Snowshoe Hares (Lepus americanus) in the Greater Yellowstone Area for Brucellosis, Tularemia, and Snowshoe Hare Virus.

    PubMed

    Tyers, Dan; Zimmer, Jeremy; Lewandowski, Kristen; Hennager, Steve; Young, John; Pappert, Ryan; Panella, Amanda; Kosoy, Olga

    2015-07-01

    We examined sera from snowshoe hares (Lepus americanus) livetrapped in the northern Greater Yellowstone Area (GYA), US, for antibodies to Brucella abortus, Francisella tularensis, and snowshoe hare virus (SSHV). Zero of 90, 0 of 67, and 40 of 100 samples were antibody positive for B. abortus, F. tularensis, and SSHV, respectively. Hares were trapped from 2009 to 2012, and of the six animals that were captured twice with at least 1 yr between captures, four developed antibody to SSHV, indicating active exposure to the agent. These findings suggest snowshoe hares in the GYA do not play a significant role as a reservoir of B. abortus, but do maintain the zoonotic, encephalitic SSHV in the population.

  14. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus)

    PubMed Central

    Nelson, Danielle D.; Duprau, Jennifer L.; Wolff, Peregrine L.; Evermann, James F.

    2016-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus). PMID:26779126

  15. Investigation of American lobster, Homarus americanus, for the presence of chlorinated dibenzo-rho-dioxins and dibenzofurans

    SciTech Connect

    Clement, R.E.; Tosine, H.M.; Taguchi, V.; Musial, C.J.; Uthe, J.F.

    1987-12-01

    The polychlorinated dibenzo-p-dioxins (PCDD) are a group of 75 compounds of current environmental concern. Most attention has been focused on the 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8,-TCDD) because of high toxicity exhibited in laboratory animal studies. Polychlorinated dibenzofurans (PCDF) comprise 135 compounds similar to the PCDD in structure and toxicity. The 2,3,7,8-tetrachlorodibenzofuran (2,3,7,8-TCDF) is the most toxic of this group. Two principal sources of PCDD and PCDF in the environment are incineration and as by-products in the manufacture and use of pentachlorophenols. American lobsters (Homarus americanus) have not been previously investigated for the presence of PCDD and PCDF. Lobsters were examined in this study because of their proximity to possible sources of PCDD and PCDF and an ability to concentrate hydrophobic organics such as polycyclic aromatic hydrocarbons (PAH) in the digestive gland.

  16. Cadmium contamination in American lobster, Homarus americanus, near a coastal lead smelter: use of multiple linear regression for management

    SciTech Connect

    Uthe, J.F.; Scott, D.P.; Chou, C.L.

    1987-04-01

    In 1980 cadmium (Cd) concentrations in digestive gland from American Lobster (Homarus americanus), captured at Belledune Harbour, New Brunswick, Canada were found to range 47.6-372 mg/kg wet weight. Since the digestive gland is commonly eaten the harbor fishery was closed. The adjacent area was decreed a controlled fishery zone. The cadmium source was a lead smelter on the harbor shore. The company installed an aqueous effluent treatment plant and improved handling procedures within the plant area. Aqueous Cd discharges were subsequently reduced by more than 95%. Cadmium levels in lobsters have been monitored annually since 1980 by sampling in early spring, immediately after the lobsters emerged from their overwintering locations. The results from the 1981-1985 monitoring program were used to model the system, analyze the data and predict the years in which certain changes to the management plan might be implemented.

  17. Serologic survey of Toxoplasma gondii in grizzly bears (Ursus arctos) and black bears (Ursus americanus), from Alaska, 1988 to 1991.

    PubMed

    Chomel, B B; Zarnke, R L; Kasten, R W; Kass, P H; Mendes, E

    1995-10-01

    We tested 644 serum samples from 480 grizzly bears and 40 black bears from Alaska (USA), collected between 1988 and 1991, for Toxoplasma gondii antibodies, using a commercially available latex agglutination test (LAT). A titer > or = 64 was considered positive. Serum antibody prevalence for T. gondii in grizzly bears (Ursus arctos) was 18% (87 of 480). Prevalence ranged from 9% (seven of 77) on Kodiak Island to 28% (15 of 54) in northern Alaska. Prevalence was directly correlated to age. No grizzly bears < 2-year-old had T. gondii antibody. High antibody titers were found mainly in grizzly bears captured north of the Arctic Circle. Antibody prevalence in black bears (Ursus americanus) from Interior Alaska was 15% (six of 40), similar to the prevalence in grizzly bears from the same area (13%; five of 40).

  18. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus).

    PubMed

    Nelson, Danielle D; Duprau, Jennifer L; Wolff, Peregrine L; Evermann, James F

    2015-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus).

  19. Serologic Survey of Snowshoe Hares (Lepus americanus) in the Greater Yellowstone Area for Brucellosis, Tularemia, and Snowshoe Hare Virus.

    PubMed

    Tyers, Dan; Zimmer, Jeremy; Lewandowski, Kristen; Hennager, Steve; Young, John; Pappert, Ryan; Panella, Amanda; Kosoy, Olga

    2015-07-01

    We examined sera from snowshoe hares (Lepus americanus) livetrapped in the northern Greater Yellowstone Area (GYA), US, for antibodies to Brucella abortus, Francisella tularensis, and snowshoe hare virus (SSHV). Zero of 90, 0 of 67, and 40 of 100 samples were antibody positive for B. abortus, F. tularensis, and SSHV, respectively. Hares were trapped from 2009 to 2012, and of the six animals that were captured twice with at least 1 yr between captures, four developed antibody to SSHV, indicating active exposure to the agent. These findings suggest snowshoe hares in the GYA do not play a significant role as a reservoir of B. abortus, but do maintain the zoonotic, encephalitic SSHV in the population. PMID:26161724

  20. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus).

    PubMed

    Nelson, Danielle D; Duprau, Jennifer L; Wolff, Peregrine L; Evermann, James F

    2015-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus). PMID:26779126

  1. Effects of Multiple Routes of Cadmium Exposure on the Hibernation Success of the American Toad (Bufo americanus)

    USGS Publications Warehouse

    James, S.M.; Little, E.E.; Semlitsch, R.D.

    2004-01-01

    The effects of multiple routes of cadmium exposure on juvenile American toads (Bufo americanus) were evaluated using environmentally relevant concentrations. During or after exposure, toads were individually hibernated for 172 days at approximately 4??C. The following experiments were conducted: (1) dermal exposure (hibernation in soil contaminated with up to 120 ??g Cd/ g (dry weight)); (2) injection exposure (single injection with cadmium to achieve a maximum whole-body nominal concentration of 3 ??g Cd/g (wet weight) 12 days before hibernation in uncontaminated soil); and, (3) oral exposure (feeding with mealworms containing ???16 ??g Cd/g (dry weight) for 50 days before hibernation in uncontaminated soil)., We hypothesized that sublethal levels of cadmium would become lethal during hibernation because of combined chemical and cold stress. No prehibernation mortality occurred in the injection and oral exposure studies. There was a significant treatment effect on whole-body cadmium concentration in toads orally or dermally exposed and on percent of cadmium retention in toads orally exposed. There was also a trend of increased time-to-burrowing and more toads partially buried with greater cadmium concentration in the dermal study, which indicated avoidance. In all 3 experiments, no significant differences were found among cadmium treatments in hibernation survival, percent of mass loss, or locomotor performance. However, toads fed mealworms averaging 4.7 ??g Cd/g (dry weight) had only 56% survival compared with 100% survival for controls. Although our results suggest that environmentally relevant levels of cadmium do not pose a great risk to American toads, factors such as soil type or prey species may increase cadmium bioavailability, and other amphibian species may be more sensitive to cadmium than B. americanus.

  2. Phylogeographic Analyses of American Black Bears (Ursus americanus) Suggest Four Glacial Refugia and Complex Patterns of Postglacial Admixture.

    PubMed

    Puckett, Emily E; Etter, Paul D; Johnson, Eric A; Eggert, Lori S

    2015-09-01

    Studies of species with continental distributions continue to identify intraspecific lineages despite continuous habitat. Lineages may form due to isolation by distance, adaptation, divergence across barriers, or genetic drift following range expansion. We investigated lineage diversification and admixture within American black bears (Ursus americanus) across their range using 22 k single nucleotide polymorphisms and mitochondrial DNA sequences. We identified three subcontinental nuclear clusters which we further divided into nine geographic regions: Alaskan (Alaska-East), eastern (Central Interior Highlands, Great Lakes, Northeast, Southeast), and western (Alaska-West, West, Pacific Coast, Southwest). We estimated that the western cluster diverged 67 ka, before eastern and Alaskan divergence 31 ka; these divergence dates contrasted with those from the mitochondrial genome where clades A and B diverged 1.07 Ma, and clades A-east and A-west diverged 169 ka. We combined estimates of divergence timing with hindcast species distribution models to infer glacial refugia for the species in Beringia, Pacific Northwest, Southwest, and Southeast. Our results show a complex arrangement of admixture due to expansion out of multiple refugia. The delineation of the genomic population clusters was inconsistent with the ranges for 16 previously described subspecies. Ranges for U. a. pugnax and U. a. cinnamomum were concordant with admixed clusters, calling into question how to order taxa below the species level. Additionally, our finding that U. a. floridanus has not diverged from U. a. americanus also suggests that morphology and genetics should be reanalyzed to assess taxonomic designations relevant to the conservation management of the species. PMID:25989983

  3. Phylogeographic Analyses of American Black Bears (Ursus americanus) Suggest Four Glacial Refugia and Complex Patterns of Postglacial Admixture.

    PubMed

    Puckett, Emily E; Etter, Paul D; Johnson, Eric A; Eggert, Lori S

    2015-09-01

    Studies of species with continental distributions continue to identify intraspecific lineages despite continuous habitat. Lineages may form due to isolation by distance, adaptation, divergence across barriers, or genetic drift following range expansion. We investigated lineage diversification and admixture within American black bears (Ursus americanus) across their range using 22 k single nucleotide polymorphisms and mitochondrial DNA sequences. We identified three subcontinental nuclear clusters which we further divided into nine geographic regions: Alaskan (Alaska-East), eastern (Central Interior Highlands, Great Lakes, Northeast, Southeast), and western (Alaska-West, West, Pacific Coast, Southwest). We estimated that the western cluster diverged 67 ka, before eastern and Alaskan divergence 31 ka; these divergence dates contrasted with those from the mitochondrial genome where clades A and B diverged 1.07 Ma, and clades A-east and A-west diverged 169 ka. We combined estimates of divergence timing with hindcast species distribution models to infer glacial refugia for the species in Beringia, Pacific Northwest, Southwest, and Southeast. Our results show a complex arrangement of admixture due to expansion out of multiple refugia. The delineation of the genomic population clusters was inconsistent with the ranges for 16 previously described subspecies. Ranges for U. a. pugnax and U. a. cinnamomum were concordant with admixed clusters, calling into question how to order taxa below the species level. Additionally, our finding that U. a. floridanus has not diverged from U. a. americanus also suggests that morphology and genetics should be reanalyzed to assess taxonomic designations relevant to the conservation management of the species.

  4. Effects of dietary supplementation of inosine monophosphate on growth performance, innate immunity and disease resistance of olive flounder (Paralichthys olivaceus).

    PubMed

    Song, Jin-Woo; Lim, Se-Jin; Lee, Kyeong-Jun

    2012-10-01

    This study was investigated to examine the effects of dietary inosine monophosphate (IMP) supplementation on growth performance, feed utilization, innate immunity, hematological parameters and disease resistance of juvenile olive flounder. Five experimental diets were formulated to contain IMP at levels of 0, 0.1, 0.2, 0.4 and 1.0%. All diets were maintained isonitrogenous (48% crude protein) and isocaloric (20.7 MJ/kg diet). Triplicate groups of olive flounder (initial body weight, 7.5 ± 0.02 g) were fed one of the experimental diets to apparent satiation (twice a day) for 14 weeks. Final body weight of fish fed 0.1-0.2% IMP were significantly higher than that of fish fed the 1.0% IMP. Groups of fish fed 0.2 or 0.4% IMP diet had significantly higher myeloperoxidase and lysozyme activities than fish fed the control diet. However, nitro-blue-tetrazolium and superoxide dismutase activities were not significantly different among all treatments. In the challenge test against Streptococcus iniae, cumulative mortality of fish fed IMP supplemented diets was significantly lower (15%, 4%, 4% and 9% for 0.1%, 0.2%, 0.4% and 1.0% IMP, respectively) than that of fish fed the control group (87%). The results suggest that IMP supplementation of 0.46-1.84 g into a kg of fish meal based diet (0.1-0.4% IMP product) can enhance innate immunity and disease resistance of olive flounder.

  5. Effects of dietary soybean stachyose and phytic acid on gene expressions of serine proteases in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Mi, Haifeng; Mai, Kangsen; Zhang, Wenbing; Wu, Chenglong; Cai, Yinghua

    2011-09-01

    Soybean stachyose (SBS) and phytic acid (PA) are anti-nutritional factors (ANF) which have deleterious effects on the growth and digestibility in fish. The present research studied the effects of dietary SBS and PA on the expression of three serine protease genes in the liver of Japanese flounder ( Paralichthys olivaceus). These genes are trypsinogen 1 (poTRY), elastase 1 (poEL) and chymotrypsinogen 1 (poCTRY). Eight artificial diets with graded levels of supplemented ANFs were formulated to 4 levels of SBS (0.00, 0.40, 0.80 and 1.50%), 4 levels of PA (0.00, 0.20, 0.40 and 0.80), respectively. Japanese flounder (initial weight 2.45 g ± 0.01 g) were fed with these diets for 10 weeks with three replications per treatment. At the end of 10 weeks, supplementation of 0.40% of dietary SBS or PA significantly increased the gene expression of poTRY and poCTRY ( P<0.05). The same level of dietary SBS significantly decreased the gene expression of poEL. In comparison with the control group (ANF-free), dietary PA (0.2% and 0.8%) significantly decreased the gene expression of poTRY, poCTRY and poEL ( P<0.05). However, excessive supplement of dietary SBS (1.5%) has no significant effects on these gene expressions ( P>0.05). These results suggested that dietary SBS and dietary PA could directly affect the serine protease genes at the transcriptional level in Japanese flounder, and these genes' expression was more sensitive to dietary PA than to SBS under the current experimental conditions.

  6. Fishing mortality in North Carolina's southern flounder fishery: direct estimates of instantaneous fishing mortality from a tag return experiment

    USGS Publications Warehouse

    Smith, William E.; Scharf, Frederick S.; Hightower, Joseph E.

    2009-01-01

    Estimation of harvest rates is often a critical component of fishery stock assessment and management. These assessments are often based on catch-at-age data sets generated over many years, but estimates of instantaneous fishing mortality (F) can also be obtained from a shorter-term tag return study. We conducted a 2-year tag return experiment to generate direct estimates of F for southern flounder Paralichthys lethostigma in a North Carolina estuary. The southern flounder supports lucrative commercial and recreational fisheries within the state and has experienced heavy fishing pressure for more than a decade. During 2005 and 2006, fish were captured and tagged with the assistance of commercial harvesters in the New River estuary. Tag returns were used to generate monthly estimates of F, which demonstrated a clear seasonal pattern that was consistent between years. Several important assumptions of the tag return model were accounted for through the use of double-tagged individuals, the distribution of both high- and standard-reward tags, and the completion of an independent controlled experiment to evaluate mortality related to tagging. Annual estimates of F exceeded the short-term management target in both years. Residual patterns suggest that the estimates may actually have been biased low, possibly due to delayed mixing of tagged fish. Thus, despite recently amended fishery regulations, F in the North Carolina southern flounder gill-net fishery still has the potential to greatly exceed targeted levels, which may delay stock recovery. Tag return studies can provide reliable (and nearly real-time) information about F and natural mortality as long as the experimental design addresses specific assumptions related to tagging-induced mortality, tag shedding, and nonreporting of tags.

  7. Immunoglobulin Tau Heavy Chain (IgT) in Flounder, Paralichthys olivaceus: Molecular Cloning, Characterization, and Expression Analyses

    PubMed Central

    Du, Yang; Tang, Xiaoqian; Zhan, Wenbin; Xing, Jing; Sheng, Xiuzhen

    2016-01-01

    Immunoglobulin tau (IgT) is a new teleost immunoglobulin isotype, and its potential function in adaptive immunity is not very clear. In the present study, the membrane-bound and secreted IgT (mIgT and sIgT) heavy chain genes were cloned for the first time and characterized in flounder (Paralichthys olivaceus), and found the nucleic acid sequence were exactly same in the Cτ1–Cτ4 constant domains of mIgT and sIgT, but different in variable regions and the C-terminus. The amino acid sequence of mIgT shared higher similarity with Bovichtus diacanthus (51.2%) and Dicentrarchus labrax (45.0%). Amino acid of flounder IgT, IgM, and IgD heavy chain was compared and the highest similarity was found between IgT Cτ1 and IgM Cμ1 (38%). In healthy flounder, the transcript levels of IgT mRNA were the highest in gill, spleen, and liver, and higher in peripheral blood leucocytes, skin, and hindgut. After infection and vaccination with Edwardsiella tarda via intraperitoneal injection and immersion, the qRT-PCR analysis demonstrated that the IgT mRNA level was significantly upregulated in all tested tissues, with similar dynamic tendency that increased firstly and then decreased, and higher in gill, skin, hindgut, liver, and stomach in immersion than in the injection group, but no significant difference existed in spleen and head kidney between immersion and injection groups. These results revealed that IgT responses could be simultaneously induced in both mucosal and systemic tissues after infection/vaccination via injection and immersion route, but IgT might play a more important role in mucosal immunity than in systemic immunity. PMID:27649168

  8. Immunoglobulin Tau Heavy Chain (IgT) in Flounder, Paralichthys olivaceus: Molecular Cloning, Characterization, and Expression Analyses.

    PubMed

    Du, Yang; Tang, Xiaoqian; Zhan, Wenbin; Xing, Jing; Sheng, Xiuzhen

    2016-01-01

    Immunoglobulin tau (IgT) is a new teleost immunoglobulin isotype, and its potential function in adaptive immunity is not very clear. In the present study, the membrane-bound and secreted IgT (mIgT and sIgT) heavy chain genes were cloned for the first time and characterized in flounder (Paralichthys olivaceus), and found the nucleic acid sequence were exactly same in the Cτ1-Cτ4 constant domains of mIgT and sIgT, but different in variable regions and the C-terminus. The amino acid sequence of mIgT shared higher similarity with Bovichtus diacanthus (51.2%) and Dicentrarchus labrax (45.0%). Amino acid of flounder IgT, IgM, and IgD heavy chain was compared and the highest similarity was found between IgT Cτ1 and IgM Cμ1 (38%). In healthy flounder, the transcript levels of IgT mRNA were the highest in gill, spleen, and liver, and higher in peripheral blood leucocytes, skin, and hindgut. After infection and vaccination with Edwardsiella tarda via intraperitoneal injection and immersion, the qRT-PCR analysis demonstrated that the IgT mRNA level was significantly upregulated in all tested tissues, with similar dynamic tendency that increased firstly and then decreased, and higher in gill, skin, hindgut, liver, and stomach in immersion than in the injection group, but no significant difference existed in spleen and head kidney between immersion and injection groups. These results revealed that IgT responses could be simultaneously induced in both mucosal and systemic tissues after infection/vaccination via injection and immersion route, but IgT might play a more important role in mucosal immunity than in systemic immunity. PMID:27649168

  9. Anesthetic Effect and Physiological Response in Olive Flounder (Paralichthys olivaceus) to Clove Oil in a Simulated Transport Experiment

    PubMed Central

    Gil, Hyun Woo; Ko, Min Gyun; Lee, Tae Ho; Park, In-Seok; Kim, Dong Soo

    2016-01-01

    The optimum concentrations of clove oil as an anesthetic for olive flounder (Paralichthys olivaceus) and the stress response of the fish to clove oil anesthesia were determined over a range of water temperatures, and investigated in a simulated transport experiment using analysis of various water and physiological parameters. While the time for induction of anesthesia decreased significantly as both the concentration of clove oil and water temperature increased, the recovery time increased significantly (P<0.05). The plasma cortisol concentration in fish at each temperature increased significantly up to 12 h following exposure (P<0.05), then decreased to 48 h (P<0.05). The DO dissolved oxygen concentrations, pH values, and the fish respiratory frequencies decreased over 6 h following exposure to clove oil in all experimental groups (P<0.05), whereas the NH4 + and CO2concentrations in all experimental groups increased up to 6 h (P<0.05). The pH values and DO concentrations increased with increasing clove oil concentration (P<0.05) in the 6 h following exposure, and the CO2 and NH4 + concentrations and the respiratory frequencies decreased with increasing clove oil concentration (P<0.05). The results of this experiment suggest that clove oil reduced the metabolic activity of olive flounder, thus reducing NH4 + excretion and O2 consumption. In conclusion, clove oil appears to be a cost-effective and efficient anesthetic that is safe for use and non-toxic to the fish and users. Its use provides the potential for improved transportation of olive flounder. PMID:27796007

  10. Molecular identification and expression analysis of natural resistance associated macrophage protein (Nramp) cDNA from Japanese flounder (Paralichthys olivaceus).

    PubMed

    Chen, Song-Lin; Wang, Zhi-Jian; Xu, Mei-Yu; Gui, Jian-Fang

    2006-03-01

    Natural resistance associated macrophage protein (Nramp) controls partially innate resistance to intracellular parasites. Its function is to enhance the ability of macrophages to kill pathogens. However, little is known about the structure and function of Nramp in lower vertebrates such as teleosts. We have recently isolated a cDNA encoding Nramp from Japanese flounder (Paralichthys olivaceus). The full-length cDNA of the Nramp is 3066 bp in length, including 224 bp 5' terminal UTR, 1662 bp encoding region and 1180 bp 3' terminal UTR. The 1662-nt open reading frame was found to code for a protein with 554 amino acid residues. Comparison of amino acid sequence indicated that Japanese flounder Nramp consists of 12 transmembrane (TM) domains. A consensus transport motif (CTM) containing 20 residues was observed between transmembrane domains 8 and 9. The deduced amino acid sequence of Japanese flounder had 77.30%, 82.71%, 82.67%, 79.64%, 80.72%, 90.97%, 91.16%, 60.14%, 71.48%, 61.69%, 72.37% identity with that of rainbow trout Nramp alpha and beta, channel catfish Nramp, fathead minnow Nramp, common carp Nramp, striped sea bass Nramp, red sea bream Nramp, mouse Nramp 1 and 2, human Nramp 1 and 2, respectively. RT-PCR indicated that Nramp transcripts were highly abundant in spleen, head kidney, abundant in intestine, liver and gill, and less abundant in heart. The level of Nramp mRNA in embryos gradually increases during embryogenesis from 4 h (8 cell stage) to 80 h (hatched stage) after fertilization.

  11. Diseases of flounder ( Platichthys Flesus L.) in the Dutch Wadden Sea, and their relation to stress factors

    NASA Astrophysics Data System (ADS)

    Dick Vethaak, A.

    In 1988 a survey was conducted on the occurrence of gross pathologies in flounder ( Platichthys flesus L.) in the Dutch Wadden Sea in relation to stress factors, especially near fresh-water drainage sluices. A total of 9608 flounder were collected from 16 sites in June and September. Diseases recorded were: skin ulcers (affecting 7.7% of fish); fin rot (2.8%; only included in the study in September); lymphocystis (3.7%); skeletal deformities (0.3%); liver nematode infections (5.0%); intestinal cysts of the parasite Glugea sp. (0.4%); and liver nodules (pre-neoplastic and neoplastic lesions) (< 0.1%). Prevalences of skin ulcers and fin rot in Wadden Sea flounder are considerably higher than in those captured off the Dutch North Sea coast or in the Elbe estuary in Germany. These high disease levels in the Wadden Sea, however, are largely due to extremely high prevalences of skin ulcers observed near the drainage sluices of Lake IJssel, indicating a localized effect. Fin rot occurs at relatively high prevalences over the whole study area, but the data should be interpreted with some caution, due to diagnostic problems. Associations with possible disease determinants were investigated and causal hypotheses formulated. In general fish seem to develop skin diseases at sites with strong salinity fluctuations. Nutritional deficiencies possibly related to strong salinity fluctuations, high bacterial loads in the environment and obstacles to fish migration through sluices are amongst the most likely factors determining the locally high levels of skin ulcers. Although not established, it is possible that pollution-related factors and contaminant bioavailability are important. The stress imposed by a combination of these factors may adversely affect the immune system, making the fish vulnerable to opportunistic bacterial infection.

  12. Spatial and diurnal distribution of invertebrate and fish fauna of a Zostera marina bed and nearby unvegetated sediments in Damariscotta River, Maine (USA)

    NASA Astrophysics Data System (ADS)

    Mattila, Johanna; Chaplin, Glen; Eilers, Michele R.; Heck, Kenneth L.; O'Neal, Jonathan P.; Valentine, John F.

    1999-06-01

    Fish, epibenthos and macroinfauna were collected in a Zostera marina bed and nearby unvegetated sediments in the estuary of the Damariscotta River, on the mid-coast of Maine. Samples of epibenthic fauna and fish were collected at low tides both during day and night, and samples of infauna at low tides during the day. The mean density of Zostera shoots in the study area was 335 m -2. Abundance and species number of fish were greater at night than during the day and greater in eelgrass beds ( Z. marina) than in unvegetated habitats. Daytime fish collections were dominated by Atlantic silversides ( Medinia medinia), while juvenile winter flounder ( Pseudopleuronectes americanus) dominated night collections. Also Zostera-associated epifaunal abundances and number of species were significantly higher at night than during the day. Mysis stenolepis, Idotea balthica and Littorina obtusata were dominant species in the epifauna samples. Of the total of 37 invertebrate species encountered, only five occurred both in the infaunal and epifaunal samples. Nineteen different taxa were collected from the benthic core samples. The most abundant invertebrate infaunal taxa were sipunculids, the polychaete Nereis virens, and oligochaetes. Infaunal invertebrate abundances and species diversity were significantly higher in eelgrass beds than in unvegetated sediments. The abundance and number of species of benthic invertebrates were also positively correlated to seagrass biomass. Community diversity values ( H') were relatively low but fit well in the general pattern of decreasing diversity towards northern latitudes.

  13. Severe Inbreeding and Small Effective Number of Breeders in a Formerly Abundant Marine Fish

    PubMed Central

    O'Leary, Shannon J.; Hice, Lyndie A.; Feldheim, Kevin A.; Frisk, Michael G.; McElroy, Anne E.; Fast, Mark D.; Chapman, Demian D.

    2013-01-01

    In contrast to freshwater fish it is presumed that marine fish are unlikely to spawn with close relatives due to the dilution effect of large breeding populations and their propensity for movement and reproductive mixing. Inbreeding is therefore not typically a focal concern of marine fish management. We measured the effective number of breeders in 6 New York estuaries for winter flounder (Pseudopleuronectes americanus), a formerly abundant fish, using 11 microsatellite markers (6–56 alleles per locus). The effective number of breeders for 1–2 years was remarkably small, with point estimates ranging from 65–289 individuals. Excess homozygosity was detected at 10 loci in all bays (FIS = 0.169–0.283) and individuals exhibited high average internal relatedness (IR; mean = 0.226). These both indicate that inbreeding is very common in all bays, after testing for and ruling out alternative explanations such as technical and sampling artifacts. This study demonstrates that even historically common marine fish can be prone to inbreeding, a factor that should be considered in fisheries management and conservation plans. PMID:23762473

  14. Adsorption of alpha-helical antifreeze peptides on specific ice crystal surface planes.

    PubMed Central

    Knight, C A; Cheng, C C; DeVries, A L

    1991-01-01

    The noncolligative peptide and glycopeptide antifreezes found in some cold-water fish act by binding to the ice surface and preventing crystal growth, not by altering the equilibrium freezing point of the water. A simple crystal growth and etching technique allows determination of the crystallographic planes where the binding occurs. In the case of elongated molecules, such as the alpha-helical peptides in this report, it also allows a deduction of the molecular alignment on the ice surface. The structurally similar antifreeze peptides from winter flounder (Pseudopleuronectes americanus) and Alaskan plaice (Pleuronectes quadritaberulatus) adsorb onto the (2021) pyramidal planes of ice, whereas the sculpin (Myoxocephalus scorpius) peptide adsorbs on (2110), the secondary prism planes. All three are probably aligned along (0112). These antifreeze peptides have 11-amino acid sequence repeats ending with a polar residue, and each repeat constitutes a distance of 16.5 A along the helix, which nearly matches the 16.7 A repeat spacing along (0112) in ice. This structural match is undoubtedly important, but the mechanism of binding is not yet clear. The suggested mechanism of growth inhibition operates through the influence of local surface curvature upon melting point and results in complete inhibition of the crystal growth even though individual antifreeze molecules bind at only one interface orientation. Images FIGURE 2 PMID:2009357

  15. Abundance exchange models of fish assemblages along the Hudson River Estuary Gradient, New York.

    PubMed

    Singkran, Nuanchan; Bain, Mark B

    2008-01-01

    The spatially explicit abundance exchange model (AEM) was built for four fish species: winter flounder (Pseudopleuronectes americanus), Atlantic silverside (Menidia menidia), eastern silvery minnow (Hybognathus regius), and striped bass (Morone saxatilis) along the Hudson River estuary gradient, New York. The fish and habitat data during 1974-1997 were used to develop and calibrate the AEM; and the fish data during 1998-2001 was used to validate the model. Preference indexes of fish species for dissolved oxygen, salinity, water temperature, and bottom substrates along the gradient were estimated; and these were used to compute habitat preference (HP) of the associated fish species. The species HP was a key variable in the AEM to quantify abundance and distribution patterns of the associated species along the gradient. The AEM could efficiently predict abundance and distribution patterns of all modeled species except striped bass. The model ability for predicting a local distribution range of a fish species with broad tolerance on changing environment like striped bass should be improved. PMID:19092189

  16. Transcriptome analysis of vertebral bone in the flounder, Paralichthys olivaceus (Teleostei, Pleuronectiformes), using Illumina sequencing.

    PubMed

    Ibaraki, Harumi; Wu, Xiaoming; Uji, Susumu; Yokoi, Hayato; Sakai, Yoshifumi; Suzuki, Tohru

    2015-12-01

    The processes underlying vertebral development in teleosts and tetrapods differ markedly in a variety of ways. At present, the molecular basis of teleost vertebral development and growth is poorly understood. Understanding vertebral development at the molecular level is important for aquaculture to prevent vertebral anomalies that can arise from a variety of factors, including excess vitamin A (all-trans retinol, VA) in the diet. To facilitate studies on teloest vertebral development, we performed transcriptome analysis of four month old flounder, Paralichthys olivaceus, vertebrae using next-generation sequencing. Expression profile obtained demonstrates that some members of the hh, bmp, fgf, wnt gene families, and their receptors, hox, pax, sox, dlx and tbx gene families and ntl, which are known to function in notochord and somite development in embryos, are expressed in the vertebrae. It was also showed that in addition to the retinoic acid receptor (Rar), the vertebrae express alcohol dehydrogenase 1 and retinal dehydrogenase 2 which convert VA to all-trans-retinoic acid (RA). The assembled contigs also included cytochrome p450 family members, which inactivate RA, as well as phosphatidylcholine-retinol O-acetyltransferase, which converts VA to all-trans-retinyl ester, a stock form of VA. These data suggest that in teleost vertebrae, expression of various signals and transcription factors which function in the notochord and somite development is maintained until adult stage, and RA metabolism and signaling are active to regulate transcription of RA-responsible genes, such as hedgehog and hox genes. This is the first transcriptome analysis of teleost fish vertebrae. PMID:26452303

  17. The use of innate immune responses as biomarkers in a programme of integrated biological effects monitoring on flounder (Platichthys flesus) from the southern North Sea

    NASA Astrophysics Data System (ADS)

    Skouras, Andreas; Broeg, Katja; Dizer, Halim; von Westernhagen, Hein; Hansen, Peter-Diedrich; Steinhagen, Dieter

    2003-10-01

    Immunological biomarkers that reflect the effects of exposure to environmental contaminants in coastal marine habitats were sought in European flounder (Platichthys flesus) from five locations in the German Bight with different anthropogenic impacts. During a 2-year period of sampling, innate immune responses were monitored from a total of 331 individual flounder of a body length of 18 to 25 cm. From the fish, plasma lysozyme, phagocytosis and respiratory burst activity of head kidney leucocytes were analysed and implemented as part of an integrated biological effects monitoring programme. As the measurements of the parameters applied here varied within wide ranges at some locations, spatial differences could not always be established, but some general trends could be drawn: plasma lysozyme activity was decreased in flounder contaminated with DDT adducts and some PCBs, while cellular functions such as phagocytosis and respiratory burst were stimulated by some chlorinated hydrocarbons. Correlation analysis also revealed connections not only between the parameters applied here and some contaminants but also with some biochemical parameters used as biomarkers in pollution monitoring: in flounder with decreased integrity of hepatocyte lysosomal membranes, immune functions also were impaired, and plasma lysozyme as well as phagocytosis activity of head kidney cells were impaired when the activity of cytochrome P450 1A was induced. The data presented here indicate that innate immune responses may be useful parameters to monitor cellular functions in a battery of biomarkers of different levels of biological organisation.

  18. Effects of CO2-induced ocean acidification on the growth of the larval olive flounder Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Kim, Kyung-Su; Shim, Jeong Hee; Kim, Suam

    2015-06-01

    It widely thought that ocean acidification processes that caused by atmospheric CO2 increase and accordingly lower seawater pH conditions might cause serious harm to marine food webs in certain ecosystems in the near future. Little is known about how marine fishes respond to reduced pH conditions. We investigated the effects of CO2 conditions on the growth of olive flounder ( Paralichthys olivaceus) larvae. Newly hatched larvae were reared at three different levels of pCO2 (574, 988 and 1297 µatm) in temperature-controlled (21 ± 0.5°C) water tanks for four weeks until metamorphosis. The experiment was repeated three times in May, June, and July 2011, and body lengths and weights were measured at the completion of each experiment. The results indicated that the body length and weight of flounder larvae significantly increased with increasing CO2 concentrations (P < 0.05). A higher daily growth rate during the early larval stage (hatching to 14 days) was found among the larvae reared in low pCO2 conditions, while a significantly lower growth rate was found among larvae in higher pCO2 water conditions. On the other hand, in the late larval stage (18 days after hatching to metamorphosis), the daily growth rate of larvae was much higher in high CO2 water. Bone density of larvae, however, decreased with increasing CO2 concentration in the water

  19. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis

    PubMed Central

    ZHANG, Hong-Mei; SU, Yan-Fang; SHI, Zhi-Yi; FU, Yuan-Shuai

    2014-01-01

    Tropomyosin (TM) plays a critical role in skeletal and cardiac muscle development and function. To assess the functional significance of α-TM in Japanese flounder (Paralichthys olivaceus) development and metamorphosis, cDNA from Japanese flounder was cloned and α-TM mRNA measured during development and metamorphosis. The full-length cDNA is 1 191 bp, including a 5'-untranslated region of 114 bp, a 3'-UTR of 222 bp, and an open reading frame of 855 bp encoding a polypeptide of 284 amino acids. Real-time quantitative PCR revealed that α-TM mRNA is initially expressed in unfertilized ovum, indicating the α-TM gene is maternal. Relatively low mRNA levels were observed in different embryonic stages. A higher level of α-TM mRNA was detected 3 days post hatching (dph), while the highest level was measured at 29 dph (metamorphic climax) after which it declined towards the end of metamorphosis. The expression of α-TM mRNA was up-regulated in thyroid hormone-treated larvae at 36 dph, but there was no marked difference at other stages when compared to control animals. After thiourea treatment, the expression of α-TM mRNA declined slightly. These data provide basic information that can be utilized in further studies into the role of α-TM in P. olivaceus development and metamorphosis. PMID:25017750

  20. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis.

    PubMed

    Zhang, Hong-Mei; Su, Yan-Fang; Shi, Zhi-Yi; Fu, Yuan-Shuai

    2014-07-01

    Tropomyosin (TM) plays a critical role in skeletal and cardiac muscle development and function. To assess the functional significance of α-TM in Japanese flounder (Paralichthys olivaceus) development and metamorphosis, cDNA from Japanese flounder was cloned and α-TM mRNA measured during development and metamorphosis. The full-length cDNA is 1 191 bp, including a 5'-untranslated region of 114 bp, a 3'-UTR of 222 bp, and an open reading frame of 855 bp encoding a polypeptide of 284 amino acids. Real-time quantitative PCR revealed that α-TM mRNA is initially expressed in unfertilized ovum, indicating the α-TM gene is maternal. Relatively low mRNA levels were observed in different embryonic stages. A higher level of α-TM mRNA was detected 3 days post hatching (dph), while the highest level was measured at 29 dph (metamorphic climax) after which it declined towards the end of metamorphosis. The expression of α-TM mRNA was up-regulated in thyroid hormone-treated larvae at 36 dph, but there was no marked difference at other stages when compared to control animals. After thiourea treatment, the expression of α-TM mRNA declined slightly. These data provide basic information that can be utilized in further studies into the role of α-TM in P. olivaceus development and metamorphosis.

  1. Effects of dietary soy isoflavones on feed intake, growth performance and digestibility in juvenile Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Mai, Kangsen; Zhang, Yanjiao; Chen, Wei; Xu, Wei; Ai, Qinghui; Zhang, Wenbing

    2012-12-01

    An 8-week feeding trial was conducted to investigate the effects of dietary soy isoflavones on feeding intake, growth performance, and digestion of juvenile Japanese flounder ( Paralichthys olivaceus). Four isonitrogenous (49% crude protein) and isoenergetic (20.1 MJ kg-1) diets were formulated to contain four graded levels of soy isoflavones, namely, 0, 1, 4 and 8 g soy isoflavones in 1 kg of diet. Each diet was randomly fed to triplicate tanks of fish (Initial average weight: 2.58 g ± 0.01 g), and each tank was stocked with 35 fish. No significant difference was observed among diets with levels of 0, 1 and 4 g kg-1 soy isoflavones in feed intake, weight gain, feed efficiency ratio (FER), proximate composition of fish whole body and apparent digestibility coefficients (ADC) of nutrients and energy ( P>0.05). However, high dietary soy isoflavones level (8 g kg-1) significantly depressed weight gain, FER, whole-body crude lipid content of fish and ADC of nutrients ( P<0.05). These results indicate that high level of dietary soy isoflavones (above 4 g kg-1) significantly depresses growth responses and FER of Japanese flounder. However, as the content of soy isoflavones in soybean meal is around 1 to 3 g kg-1, the adverse effects might be neglected when soybean products are used as a fish feed ingredient.

  2. Cloning and stage-specific expression of CK-M1 gene during metamorphosis of Japanese flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Chen, Yanjie; Zhang, Quanqi; Qi, Jie; Wang, Zhigang; Wang, Xubo; Sun, Yeying; Zhong, Qiwang; Li, Shuo; Li, Chunmei

    2010-05-01

    The symmetrical body of flatfish larvae changes dramatically into an asymmetrical form after metamorphosis. The molecular mechanisms responsible for this change are poorly understood. As an initial step to clarify these mechanisms, we used representational difference analysis of cDNA for the identification of genes active during metamorphosis in the Japanese flounder, Paralichthys olicaceus. One of the up-regulated genes was identified as creatine kinase muscle type 1 (CK-M1). Sequence analysis of CK-M1 revealed that it spanned 1 708 bp and encoded a protein of 382 amino acids. The overall amino acid sequence of the CK-M1 was highly conserved with those of other organisms. CK-M1 was expressed in adult fish tissues, including skeletal muscle, intestine and gill. Whole mount in-situ hybridization showed that the enhanced expression of CK-M1 expanded from the head to the whole body of larvae as metamorphosis progressed. Quantitative analysis revealed stage-specific high expression of CK-M1 during metamorphosis. The expression level of CK-M1 increased initially and peaked at metamorphosis, decreased afterward, and finally returned to the pre-metamorphosis level. This stage-specific expression pattern suggested strongly that CK-M1 was related to metamorphosis in the Japanese flounder. Its specific role in metamorphosis requires further study.

  3. Winter winds and river discharge determine juvenile southern flounder ( Paralichthys lethostigma) recruitment and distribution in North Carolina estuaries

    NASA Astrophysics Data System (ADS)

    Taylor, J. Christopher; Miller, John M.; Pietrafesa, Leonard J.; Dickey, David A.; Ross, Steve W.

    2010-07-01

    Retrospective analyses of a 23 year data set on abundance of Age 0 southern flounder in 105 estuarine nursery areas in the coastal region of North Carolina showed that discernible temporal and spatial patterns exist among clusters of stations. Furthermore, these patterns could be quantitatively related to certain meteorological and hydrological variables, namely winds from the east-southeast (E-SE) and from the north-northeast (N-NE) sectors and river runoff, which explained up to 83% of the interannual variability in numbers. We developed a regression model using recent catch data (1987-2002) and used the model to hindcast an earlier segment of the time series (1979-1986). The model was found to be quite robust, and could predict year class strength within 1 to 80% in the test set of data. We interpret these results to mean that hydrodynamic factors are principally responsible for the observed interannual recruitment variability in southern flounder in NC, since the interannual pattern in abundance of Age 0 fish persists for 2 more years of adult life. Finally, we discuss the implications of the variable spatial distribution patterns for estimates of year class strength from juvenile abundance data. It is possible that estimates of year class strength with a useful level of confidence could be obtained from meteorological data during the larval migration period.

  4. Ocular-side lateralization of adult-type chromatophore precursors: development of pigment asymmetry in metamorphosing flounder larvae.

    PubMed

    Washio, Youhei; Aritaki, Masato; Fujinami, Yuichiro; Shimizu, Daisuke; Yokoi, Hayato; Suzuki, Tohru

    2013-05-01

    The adult-type chromatophores of flounder differentiate at metamorphosis in the skin of ocular side to establish asymmetric pigmentation. In young larva and before metamorphosis, adult-type melanophores that migrate to the ocular side during metamorphosis reside at the base of the dorsal fin as latent precursors. However, the migration route taken by these precursor cells and the mechanisms by which lateralization and asymmetric pigmentation develop on the ocular side are unknown. To further investigate this migration and lateralization, we used in situ hybridization with gch2 probe, a marker for melanoblasts and xanthoblasts (precursors of adult type chromatophores), to examine the distribution of chromatophore precursors in metamorphosing larvae. The gch2-positive precursors were present in the myoseptum as well as in the skin. This finding indicated that these precursors migrated from the dorsal part of the fin to the skin via the myoseptum. Additionally, there were much fewer gch2-positive cells in the myoseptum of the blind side than in the skin and myoseptum of the ocular side, and this finding indicated either that migration of the precursor cells into the myoseptum of blind side was inhibited or that the precursors were eliminated from the myoseptum of the blind side. Therefore, we propose that the signals responsible for development of asymmetric pigmentation in flounder reside not only in the skin but on a larger scale and in multiple tissues throughout the lateral half of the trunk.

  5. Tissue-specific bioaccumulation and oxidative stress responses in juvenile Japanese flounder ( Paralichthys olivaceus) exposed to mercury

    NASA Astrophysics Data System (ADS)

    Huang, Wei; Cao, Liang; Ye, Zhenjiang; Lin, Longshan; Chen, Quanzhen; Dou, Shuozeng

    2012-07-01

    To understand mercury (Hg) toxicity in marine fish, we measured Hg accumulation in juvenile Japanese flounder ( Paralichthys olivaceus) and assessed the effects on growth and antioxidant responses. After Hg exposure (control, 5, 40, and 160 μg/L Hg) for 28 d, fish growth was significantly reduced. The accumulation of Hg in fish was dose-dependent and tissue-specific, with the maximum accumulation in kidney and liver, followed by gills, bone, and muscle. Different antioxidants responded differently to Hg exposure to cope with the induction of lipid peroxidation (LPO), which was also tissue-specific and dosedependent. As Hg concentration increased, superoxide dismutase (SOD) and catalase (CAT) activities increased significantly, whereas glutathione S -transferase (GST) activity and glutathione (GSH) levels decreased significantly in the gills. SOD and glutathione peroxidase (GPx) activities and the GSH level increased significantly in the liver. SOD activity and GSH levels increased significantly, but CAT activity decreased significantly with an increase in Hg concentration in the kidney. LPO was induced significantly by elevated Hg in the gills and kidney but was least affected in the liver. Therefore, oxidative stress biomarkers in gills were more sensitive than those in the liver and kidney to Hg exposure. Thus, the gills have potential as bioindicators for evaluating Hg toxicity in juvenile flounder.

  6. A combined measurement of metal bioaccumulation and condition indices in juvenile European flounder, Platichthys flesus, from European estuaries.

    PubMed

    Kerambrun, E; Henry, F; Cornille, V; Courcot, L; Amara, R

    2013-04-01

    Condition indices and metal bioaccumulation of early life stages of juvenile flounder (5-10 cm) were determined in three anthropogenic estuaries (the Scheldt, Seine and Loire) and compared to a reference site (the Canche). Significant correlations were found between metal concentrations in sediment and (i) fish liver for Cd, Cu, Ni, Pb, V and Zn and (ii) fish gills for Cd and Mn. Metal accumulation in juvenile flounder from the three anthropogenic estuaries coincided with significantly lower Fulton's K indices (from 0.99 ± 0.03 to 1.06 ± 0.01 mg mm(-3)) compared to those from the Canche estuary (from 1.02 ± 0.01 to 1.13 ± 0.01 mg mm(-3)). This discrepancy in fish condition index increased with fish size and therefore, strongly depends on the time juvenile spend in estuary. Muscle lipid contents and Triacylglycerol to Sterol ratios were significantly lower in fish collected in the Scheldt (lipid content: 21.3 ± 3.6%), Seine (17.9 ± 19.8%) and Loire (19.5 ± 2.4%) estuaries compared to those originating from the Canche (38.3 ± 4.6%). This study highlights that combined measures of both fish metal contents and condition indices gives a relevant assessment of juvenile fish health growing in anthropogenic estuaries.

  7. Early contamination of European flounder (Platichthys flesus) by PCDD/Fs and dioxin-like PCBs in European waters.

    PubMed

    Nunes, Margarida; Martinho, Filipe; Vernisseau, Anaïs; Marchand, Philippe; Le Bizec, Bruno; van der Veer, Henk W; Cabral, Henrique N; Ramos, Fernando; Pardal, Miguel A

    2014-08-15

    Contamination levels and profiles of 7 polychlorinated-p-dioxins, 10 polychlorinated furans (PCDD/Fs) and 12 dioxin-like polychlorinated biphenyls (dl-PCBs) were investigated in juvenile European flounder (Platichthys flesus) captured in different nursery areas in the northeastern Atlantic coast across its geographical distribution range. The toxic equivalent concentrations (WHO-TEQfish) were also determined in order to evaluate which P. flesus population was more exposed to dioxin-like toxicity. Juveniles caught in the Sørfjord (Norway) showed the lowest WHO-TEQfish concentration (0.052 pg WHO-TEQfish g(-1)wet weight) whereas the highest value was observed in fish from the Wadden Sea (The Netherlands; 0.291 pg WHO-TEQfish g(-1)ww), mainly due to the greater contribution of 2,3,7,8-tetrachlorodibenzo-p-dioxin, the most toxic congener. Nonetheless, when comparing the results with existent tissue residue-based toxicity benchmarks, no adverse effects resulting from PCDD/Fs and dl-PCBs are expected to occur in flounder from the studied systems.

  8. An outbreak of VHSV (viral hemorrhagic septicemia virus) infection in farmed Japanese flounder Paralichthys olivaceus in Japan.

    PubMed

    Isshik, T; Nishizawa, T; Kobayashi, T; Nagano, T; Miyazaki, T

    2001-11-01

    A rhabdoviral disease occurred in farmed populations of market sized Japanese flounder (hirame) Paralichthys olivaceus in the Seto Inland Sea of Japan in 1996. The causative agent was identified as viral hemorrhagic septicemia virus (VHSV) based on morphological, immunological, and genetic analyses. Diseased fish that were artificially injected with a representative virus isolate showed the same pathological signs and high mortality as observed in the natural outbreak. This is the first report of an outbreak of VHSV infection in cultured fish in Japan. Clinical signs of diseased fish included dark body coloration, an expanded abdomen due to ascites, congested liver, splenomegaly, and a swollen kidney. Myocardial necrosis was most prominent and accompanied by inflammatory reactions. Necrotic lesions also occurred in the liver, spleen and hematopoietic tissue, and were accompanied by circulatory disturbances due to cardiac failure. Hemorrhagic lesions did not always appear in the lateral musculature. Transmission electron microscopy revealed many rhabdovirus particles and associated inclusion bodies containing nucleocapsids in the necrotized myocardium. The histopathological findings indicated that the necrotizing myocarditis could be considered a pathognomonic sign of VHSV infection in Japanese flounder. PMID:11775799

  9. Commercial fishing gear modifications to reduce interactions between Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) and the southern flounder (Paralichthys lethostigma) fishery in North Carolina (USA).

    PubMed

    Levesque, Juan C; Hager, Christian; Diaddorio, Eric; Dickey, R Jason

    2016-01-01

    Bycatch of protected species in commercial fishing operations is a primary concern to fishery managers because it threatens the conservation, protection, and recovery of fragile species, such as the Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus). One potential solution to reduce the risk associated with commercial fishing operations is to design commercial fishing gear that is more selective in terms of interactions between Atlantic sturgeon and commercial fisheries. Given this conservation and management need, the overarching goal was to reduce Atlantic sturgeon fishery interactions and maintain southern flounder (Paralichthys lethostigma) catch in North Carolina. The specific objectives of this study were to design and evaluate the effectiveness of a modified gillnet. Overall, the results proved that lowering the profile and amount of webbing had a beneficial impact at reducing Atlantic sturgeon incidental encounters and bycatch. The modified gillnet reduced bycatch and Atlantic sturgeon encounters by 39.6% and 60.9%, respectively. Our design entangled 51.6% fewer southern flounder, which corresponded to a 48.9% reduction in total weight; the modified gear entangled slightly larger southern flounder than the control gear. Our findings showed the number of Atlantic sturgeon encounters was positively associated with mean water depth, with more Atlantic sturgeon encountered in deeper (5.1-6.3 m) than shallower waters; 75% were encountered at depths between 4.6 and 6.1 m. Most southern flounder (n = 518, 39.7%) were taken at a water depth between 3.76 and 5.0 m. This observation suggests that southern flounder prefer slightly shallower waters than Atlantic sturgeon. PMID:27547524

  10. Commercial fishing gear modifications to reduce interactions between Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) and the southern flounder (Paralichthys lethostigma) fishery in North Carolina (USA).

    PubMed

    Levesque, Juan C; Hager, Christian; Diaddorio, Eric; Dickey, R Jason

    2016-01-01

    Bycatch of protected species in commercial fishing operations is a primary concern to fishery managers because it threatens the conservation, protection, and recovery of fragile species, such as the Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus). One potential solution to reduce the risk associated with commercial fishing operations is to design commercial fishing gear that is more selective in terms of interactions between Atlantic sturgeon and commercial fisheries. Given this conservation and management need, the overarching goal was to reduce Atlantic sturgeon fishery interactions and maintain southern flounder (Paralichthys lethostigma) catch in North Carolina. The specific objectives of this study were to design and evaluate the effectiveness of a modified gillnet. Overall, the results proved that lowering the profile and amount of webbing had a beneficial impact at reducing Atlantic sturgeon incidental encounters and bycatch. The modified gillnet reduced bycatch and Atlantic sturgeon encounters by 39.6% and 60.9%, respectively. Our design entangled 51.6% fewer southern flounder, which corresponded to a 48.9% reduction in total weight; the modified gear entangled slightly larger southern flounder than the control gear. Our findings showed the number of Atlantic sturgeon encounters was positively associated with mean water depth, with more Atlantic sturgeon encountered in deeper (5.1-6.3 m) than shallower waters; 75% were encountered at depths between 4.6 and 6.1 m. Most southern flounder (n = 518, 39.7%) were taken at a water depth between 3.76 and 5.0 m. This observation suggests that southern flounder prefer slightly shallower waters than Atlantic sturgeon.

  11. Commercial fishing gear modifications to reduce interactions between Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) and the southern flounder (Paralichthys lethostigma) fishery in North Carolina (USA)

    PubMed Central

    Hager, Christian; Diaddorio, Eric; Dickey, R. Jason

    2016-01-01

    Bycatch of protected species in commercial fishing operations is a primary concern to fishery managers because it threatens the conservation, protection, and recovery of fragile species, such as the Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus). One potential solution to reduce the risk associated with commercial fishing operations is to design commercial fishing gear that is more selective in terms of interactions between Atlantic sturgeon and commercial fisheries. Given this conservation and management need, the overarching goal was to reduce Atlantic sturgeon fishery interactions and maintain southern flounder (Paralichthys lethostigma) catch in North Carolina. The specific objectives of this study were to design and evaluate the effectiveness of a modified gillnet. Overall, the results proved that lowering the profile and amount of webbing had a beneficial impact at reducing Atlantic sturgeon incidental encounters and bycatch. The modified gillnet reduced bycatch and Atlantic sturgeon encounters by 39.6% and 60.9%, respectively. Our design entangled 51.6% fewer southern flounder, which corresponded to a 48.9% reduction in total weight; the modified gear entangled slightly larger southern flounder than the control gear. Our findings showed the number of Atlantic sturgeon encounters was positively associated with mean water depth, with more Atlantic sturgeon encountered in deeper (5.1–6.3 m) than shallower waters; 75% were encountered at depths between 4.6 and 6.1 m. Most southern flounder (n = 518, 39.7%) were taken at a water depth between 3.76 and 5.0 m. This observation suggests that southern flounder prefer slightly shallower waters than Atlantic sturgeon. PMID:27547524

  12. Identification and characterization of a cDNA encoding a crustin-like, putative antibacterial protein from the American lobster Homarus americanus.

    PubMed

    Christie, Andrew E; Rus, Szymon; Goiney, Christopher C; Smith, Christine M; Towle, David W; Dickinson, Patsy S

    2007-07-01

    Pathogenic challenges in decapod crustaceans are combated by innate immune responses, including the production and secretion of soluble antibacterial proteins into the hemolymph. Among the antibacterials that have been identified in decapod species are the crustins, a group of four-disulfide core/whey-acidic-protein (WAP) domain-containing proteins, which target marine/salt tolerant Gram-positive bacteria. To begin to assess the possible role of crustins in combating bacterial invasion in the American lobster Homarus americanus, we identified and sequenced a 744 base pair cDNA that encodes a novel 96 amino acid crustin-like protein. Comparison of H. americanus crustin (Hoa-crustin) with crustins from other decapod species showed that it is most similar to an isoform predicted from the European lobster Homarus gammarus ( approximately 86% identity). With our identification of the Hoa-crustin cDNA, we are positioned to begin molecular and physiological investigations of the regulation and function of this putative antibacterial protein in H. americanus. PMID:17418897

  13. Embryonic and larval development in barfin flounder Verasper moseri (Jordan and Gilbert)

    NASA Astrophysics Data System (ADS)

    Du, Rongbin; Wang, Yongqiang; Jiang, Haibin; Liu, Liming; Wang, Maojian; Li, Tianbao; Zhang, Shubao

    2010-01-01

    side on day 50, when the average body length attained 2.5±0.18 cm, and juveniles accomplished metamorphosis to young. The embryonic and larval characters of several flounder species are compared.

  14. rRNA operons and genome size of 'Candidatus Liberibacter americanus', a bacterium associated with citrus huanglongbing in Brazil.

    PubMed

    Wulff, N A; Eveillard, S; Foissac, X; Ayres, A J; Bové, J-M

    2009-08-01

    Huanglongbing is one of the most severe diseases of citrus worldwide and is associated with 'Candidatus (Ca.) Liberibacter africanus' in Africa, 'Ca. Liberibacter asiaticus' in Asia and the Americas (Brazil, USA and Cuba) and 'Ca. Liberibacter americanus' (Lam) in Brazil. In the absence of axenic cultures, genetic information on liberibacters is scarce. The sequences of the entire 23S rRNA and 5S rRNA genes from Lam have now been obtained, using a consensus primer designed on known tRNAMet sequences of rhizobia. The size of the Lam genome was determined by PFGE, using Lam-infected periwinkle plants for bacterial enrichment, and was found to be close to 1.31 Mbp. In order to determine the number of ribosomal operons on the Lam genome, probes designed to detect the 16S rRNA gene and the 3' end of the 23S rRNA gene were developed and used for Southern hybridization with I-CeuI-treated genomic DNA. Our results suggest that there are three ribosomal operons in a circular genome. Lam is the first liberibacter species for which such data are available.

  15. Forces generated during stretch in the heart of the lobster Homarus americanus are anisotropic and are altered by neuromodulators.

    PubMed

    Dickinson, E S; Johnson, A S; Ellers, O; Dickinson, P S

    2016-04-15

    Mechanical and neurophysiological anisotropies mediate three-dimensional responses of the heart of ITALIC! Homarus americanus Although hearts ITALIC! in vivoare loaded multi-axially by pressure, studies of invertebrate cardiac function typically use uniaxial tests. To generate whole-heart length-tension curves, stretch pyramids at constant lengthening and shortening rates were imposed uniaxially and biaxially along longitudinal and transverse axes of the beating whole heart. To determine whether neuropeptides that are known to modulate cardiac activity in ITALIC! H. americanusaffect the active or passive components of these length-tension curves, we also performed these tests in the presence of SGRNFLRFamide (SGRN) and GYSNRNYLRFamide (GYS). In uniaxial and biaxial tests, both passive and active forces increased with stretch along both measurement axes. The increase in passive forces was anisotropic, with greater increases along the longitudinal axis. Passive forces showed hysteresis and active forces were higher during lengthening than shortening phases of the stretch pyramid. Active forces at a given length were increased by both neuropeptides. To exert these effects, neuropeptides might have acted indirectly on the muscle via their effects on the cardiac ganglion, directly on the neuromuscular junction, or directly on the muscles. Because increases in response to stretch were also seen in stimulated motor nerve-muscle preparations, at least some of the effects of the peptides are likely peripheral. Taken together, these findings suggest that flexibility in rhythmic cardiac contractions results from the amplified effects of neuropeptides interacting with the length-tension characteristics of the heart.

  16. Aerococcus viridans expression of Cpn60 is associated with virulence during infection of the American lobster, Homarus americanus Milne Edwards.

    PubMed

    Clark, K F; Greenwood, S J

    2011-11-01

    The Gram-positive bacterium Aerococcus viridans var. homari is a well-documented causative agent of the lethal systemic disease gaffkemia in both the American lobster, Homarus americanus, and the European lobster, Homarus gammarus. Previous phenotypic characterization has been unsuccessful at differentiating avirulent from virulent strains without performing lethal animal infection trials. Recent genetic characterization of A. viridans strains through 16S rRNA sequencing and random amplification of polymorphic DNA fingerprinting has revealed the presence of two subtypes. However, subtype 1 contains both virulent and avirulent strains which are genetically identical. The purpose of this study was to determine the proteomic mediators of virulence in A. viridans. Quantitative proteomic mapping of these two strains has revealed 29 differentially expressed protein spots, seven of which are only expressed in the virulent strain and could act as virulence factors. One protein, chaperonin 60 (Cpn60), is uniquely expressed in the virulent strain and has been shown to act as a virulence factor in many other bacteria. The proteomic mapping strategy employed in this study is the first to show phenotypic differences between virulent and avirulent strains. Cpn60 expression represents a potentially useful tool for identifying the virulent strains of A. viridans in epidemiological studies. PMID:21988355

  17. Cold-adapted digestive aspartic protease of the clawed lobsters Homarus americanus and Homarus gammarus: biochemical characterization.

    PubMed

    Rojo, Liliana; García-Carreño, Fernando; de Los Angeles Navarrete del Toro, Maria

    2013-02-01

    Aspartic proteinases in the gastric fluid of clawed lobsters Homarus americanus and Homarus gammarus were isolated to homogeneity by single-step pepstatin-A affinity chromatography; such enzymes have been previously identified as cathepsin D-like enzymes based on their deduced amino acid sequence. Here, we describe their biochemical characteristics; the properties of the lobster enzymes were compared with those of its homolog, bovine cathepsin D, and found to be unique in a number of ways. The lobster enzymes demonstrated hydrolytic activity against synthetic and natural substrates at a wider range of pH; they were more temperature-sensitive, showed no changes in the K(M) value at 4°C, 10°C, and 25°C, and had 20-fold higher k(cat)/K(M) values than bovine enzyme. The bovine enzyme was temperature-dependent. We propose that both properties arose from an increase in molecular flexibility required to compensate for the reduction of reaction rates at low habitat temperatures. This is supported by the fast denaturation rates induced by temperature. PMID:22648335

  18. 1-/sup 14/C-n-hexadecane disposition in the spiny lobster, Panulirus argus and the American lobster, Homarus americanus

    SciTech Connect

    Little, P.J.; James, M.O.; Foureman, G.L.; Weatherby, R.P.; Bend, J.R.

    1986-05-01

    1-/sup 14/C-n-hexadecane, a model compound for the non-volatile aliphatic hydrocarbon components of crude oil, was administered by intrapericardial injection to the spiny lobster, Panulirus argus, and the clawed or American lobster, Homarus americanus. Experiments were conducted in Florida (spiny lobster) and Maine (American lobster). The animals were sacrificed at various times from 0.5 hr to 8 wks after administration of the dose. The tissues and fluids were analyzed for /sup 14/C content by digestion or catalytic oxidation and liquid scintillation counting. Selected tissues (hepatopancreas, tail muscle and hemolymph) were extracted with ethyl acetate to allow quantitation of the unmetabolized n-hexadecane by thin layer chromatography. n-Hexadecane-derived radioactivity was very persistent in both the spiny lobster (t1/2 = 4.6 wk) and the American lobster (t1/2 = 11.2 wk). In both lobsters, the hepatopancreas (HP) acquired the highest specific activity and the tail muscle had the longest half life for elimination from an individual tissue. Although hexadecane was metabolized more rapidly in the HP of the spiny lobster than in the HP of the American lobster, unmetabolized hexadecane persisted in the HPs of both species for at least 8 weeks after the dose (the longest time studied).

  19. Copper-metallothioneins in the American lobster, Homarus americanus: potential role as Cu(I) donors to apohemocyanin

    SciTech Connect

    Brouwer, M.; Whaling, P.; Engel, D.W.

    1986-03-01

    The physiological function of copper(I)-metallothionein is not well understood. The respiratory function of hemocyanin, a copper(I)-containing respiratory protein found in the hemolymph of many invertebrates, has been known a long time. However, the mechanism by which Cu(I) is inserted into the oxygen-binding site of apohemocyanin is completely unknown. This investigation tests that hypothesis that copper(I)-metallothionein may act as a Cu(I) donor to apohemocyanin. To this end, copper-binding proteins and hemocyanin were purified from the digestive gland and hemolymph of the American lobster, Homarus americanus. In the presence of ..beta..-mercaptoethanol, the copper-binding proteins can be resolved into three components of DEAE-cellulose. The first two have been characterized as metallothioneins. The cysteine content of the third component is half of that of components I and II. The purified proteins are not capable of transferring Cu(I) to the active sites of completely copper-free apohemocyanin. They are capable, however, of transferring Cu(I) to active sites of hemocyanin containing reduced amounts of Cu(I), suggesting that the conformational state of hemocyanin is the determining factor in the Cu(I) transfer mechanism.

  20. Magnetic resonance imaging diagnosis of intervertebral disc disease and myelomalacia in an American black bear (Ursus americanus).

    PubMed

    Knafo, S Emmanuelle; Divers, Stephen J; Rech, Raquel; Platt, Simon R

    2012-06-01

    A 23-yr-old black bear (Ursus americanus) was examined because of paralysis of unknown duration. The precise onset of clinical signs was unknown as a result of seasonal torpor. The bear was immobilized and transported to a university veterinary teaching hospital for further evaluation and treatment. Radiography revealed increased mineral opacity and ventral bridging across vertebral segments T8-11. Magnetic resonance imaging demonstrated dorsal and ventral compression of the spinal cord at T8-9. Given the bear's advanced age, the unknown duration of spinal cord compression, unknown presence of deep pain perception, and thus an unknown prognosis for surgical success, euthanasia was elected. Postmortem examination revealed severe spondylosis deformans from T7 to L3 and dorsal extradural extruded disc material in the area of T8-9. Histopathology demonstrated the dorsal horns of the spinal cord at T9 were replaced by foamy macrophages extending into the dorsal and lateral funiculi of the white matter compatible with focal, severe, chronic myelomalacia. This is the first report of intervertebral disc disease and myelomalacia diagnosed using MRI in a large carnivore. PMID:22779249

  1. The effects of hibernation and captivity on glucose metabolism and thyroid hormones in American black bear (Ursus americanus).

    PubMed

    McCain, Stephanie; Ramsay, Ed; Kirk, Claudia

    2013-06-01

    American black bears (Ursus americanus) have been shown to become transiently insulin resistant and hypothyroid during winter, but no studies have investigated these changes in long-term captive bears or in bears which remain awake year-round. Wild, captive hibernating, and captive nonhibernating bears were evaluated at times corresponding to three of their major physiologic stages: fall (hyperphagic stage), winter (hibernation stage), and summer (normal activity stage). Combined insulin and glucose tolerance tests and thyroid hormone profiles were performed on all bears during each stage. All three groups of bears had evidence of insulin resistance during the winter, as compared to the summer or fall, based on glucose tolerance curves. Analysis of thyroid hormone concentration varied and distinct patterns or similarities were not apparent. While obesity in captive American black bears is multifactorial, the finding that, regardless of their ability to hibernate, captive bears retain similar physiology to their wild counterparts indicates that captive bears' complex physiologic changes need to be addressed in their management. PMID:23805551

  2. Identification and phylogenetic analysis of Dirofilaria ursi (Nematoda: Filarioidea) from Wisconsin black bears (Ursus americanus) and its Wolbachia endosymbiont.

    PubMed

    Michalski, Michelle L; Bain, Odile; Fischer, Kerstin; Fischer, Peter U; Kumar, Sanjay; Foster, Jeremy M

    2010-04-01

    Dirofilaria ursi is a filarial nematode of American black bears (Ursus americanus Pallas, 1780) that is vectored by black flies (Simuliidae) in many parts of the United States. In northwestern Wisconsin, the prevalence of microfilaremic bears during the fall hunting season was 21% (n = 47). Unsheathed blood microfilariae from Wisconsin bears possess characters consistent with the original description of D. ursi, as do adult worms observed histologically and grossly. Immunohistochemistry was used to identify the Wolbachia endosymbiont in the hypodermis and lateral cords of an adult female D. ursi. Amplification of wsp, gatB, coxA, fbpA, and ftsZ bacterial sequences from parasite DNA confirmed the presence of Wolbachia, and molecular phylogenetic analysis of the Wolbachia ftsZ gene groups the endosymbiont with Wolbachia from D. immitis and D. repens. Phylogenetic analysis of D. ursi 5s rDNA sequence confirms the morphological observations grouping this parasite as a member of Dirofilaria, and within the Dirofilaria - Onchocerca clade of filarial nematodes. This is the first report of Wolbachia characterization and molecular phylogeny information for D. ursi. PMID:19916630

  3. Grizzly bears (Ursus arctos horribilis) and black bears (Ursus americanus) prevent trabecular bone loss during disuse (hibernation).

    PubMed

    McGee-Lawrence, Meghan E; Wojda, Samantha J; Barlow, Lindsay N; Drummer, Thomas D; Castillo, Alesha B; Kennedy, Oran; Condon, Keith W; Auger, Janene; Black, Hal L; Nelson, O Lynne; Robbins, Charles T; Donahue, Seth W

    2009-12-01

    Disuse typically causes an imbalance in bone formation and bone resorption, leading to losses of cortical and trabecular bone. In contrast, bears maintain balanced intracortical remodeling and prevent cortical bone loss during disuse (hibernation). Trabecular bone, however, is more detrimentally affected than cortical bone in other animal models of disuse. Here we investigated the effects of hibernation on bone remodeling, architectural properties, and mineral density of grizzly bear (Ursus arctos horribilis) and black bear (Ursus americanus) trabecular bone in several skeletal locations. There were no differences in bone volume fraction or tissue mineral density between hibernating and active bears or between pre- and post-hibernation bears in the ilium, distal femur, or calcaneus. Though indices of cellular activity level (mineral apposition rate, osteoid thickness) decreased, trabecular bone resorption and formation indices remained balanced in hibernating grizzly bears. These data suggest that bears prevent bone loss during disuse by maintaining a balance between bone formation and bone resorption, which consequently preserves bone structure and strength. Further investigation of bone metabolism in hibernating bears may lead to the translation of mechanisms preventing disuse-induced bone loss in bears into novel treatments for osteoporosis.

  4. Grizzly bears (Ursus arctos horribilis) and black bears (Ursus americanus) prevent trabecular bone loss during disuse (hibernation).

    PubMed

    McGee-Lawrence, Meghan E; Wojda, Samantha J; Barlow, Lindsay N; Drummer, Thomas D; Castillo, Alesha B; Kennedy, Oran; Condon, Keith W; Auger, Janene; Black, Hal L; Nelson, O Lynne; Robbins, Charles T; Donahue, Seth W

    2009-12-01

    Disuse typically causes an imbalance in bone formation and bone resorption, leading to losses of cortical and trabecular bone. In contrast, bears maintain balanced intracortical remodeling and prevent cortical bone loss during disuse (hibernation). Trabecular bone, however, is more detrimentally affected than cortical bone in other animal models of disuse. Here we investigated the effects of hibernation on bone remodeling, architectural properties, and mineral density of grizzly bear (Ursus arctos horribilis) and black bear (Ursus americanus) trabecular bone in several skeletal locations. There were no differences in bone volume fraction or tissue mineral density between hibernating and active bears or between pre- and post-hibernation bears in the ilium, distal femur, or calcaneus. Though indices of cellular activity level (mineral apposition rate, osteoid thickness) decreased, trabecular bone resorption and formation indices remained balanced in hibernating grizzly bears. These data suggest that bears prevent bone loss during disuse by maintaining a balance between bone formation and bone resorption, which consequently preserves bone structure and strength. Further investigation of bone metabolism in hibernating bears may lead to the translation of mechanisms preventing disuse-induced bone loss in bears into novel treatments for osteoporosis. PMID:19703606

  5. First evidence for slave rebellion: enslaved ant workers systematically kill the brood of their social parasite protomognathus americanus.

    PubMed

    Achenbach, Alexandra; Foitzik, Susanne

    2009-04-01

    During the process of coevolution, social parasites have evolved sophisticated strategies to exploit the brood care behavior of their social hosts. Slave-making ant queens invade host colonies and kill or eject all adult host ants. Host workers, which eclose from the remaining brood, are tricked into caring for the parasite brood. Due to their high prevalence and frequent raids, following which stolen host broods are similarly enslaved, slave-making ants exert substantial selection upon their hosts, leading to the evolution of antiparasite adaptations. However, all host defenses shown to date are active before host workers are parasitized, whereas selection was thought to be unable to act on traits of already enslaved hosts. Yet, here we demonstrate the rebellion of enslaved Temnothorax workers, which kill two-thirds of the female pupae of the slave-making ant Protomognathus americanus. Thereby, slaves decrease the long-term parasite impact on surrounding related host colonies. This novel antiparasite strategy of enslaved workers constitutes a new level in the coevolutionary battle after host colony defense has failed. Our discovery is analogous to recent findings in hosts of avian brood parasites where perfect mimicry of parasite eggs leads to the evolution of chick recognition as a second line of defense.

  6. The effects of hibernation and captivity on glucose metabolism and thyroid hormones in American black bear (Ursus americanus).

    PubMed

    McCain, Stephanie; Ramsay, Ed; Kirk, Claudia

    2013-06-01

    American black bears (Ursus americanus) have been shown to become transiently insulin resistant and hypothyroid during winter, but no studies have investigated these changes in long-term captive bears or in bears which remain awake year-round. Wild, captive hibernating, and captive nonhibernating bears were evaluated at times corresponding to three of their major physiologic stages: fall (hyperphagic stage), winter (hibernation stage), and summer (normal activity stage). Combined insulin and glucose tolerance tests and thyroid hormone profiles were performed on all bears during each stage. All three groups of bears had evidence of insulin resistance during the winter, as compared to the summer or fall, based on glucose tolerance curves. Analysis of thyroid hormone concentration varied and distinct patterns or similarities were not apparent. While obesity in captive American black bears is multifactorial, the finding that, regardless of their ability to hibernate, captive bears retain similar physiology to their wild counterparts indicates that captive bears' complex physiologic changes need to be addressed in their management.

  7. Forces generated during stretch in the heart of the lobster Homarus americanus are anisotropic and are altered by neuromodulators.

    PubMed

    Dickinson, E S; Johnson, A S; Ellers, O; Dickinson, P S

    2016-04-15

    Mechanical and neurophysiological anisotropies mediate three-dimensional responses of the heart of ITALIC! Homarus americanus Although hearts ITALIC! in vivoare loaded multi-axially by pressure, studies of invertebrate cardiac function typically use uniaxial tests. To generate whole-heart length-tension curves, stretch pyramids at constant lengthening and shortening rates were imposed uniaxially and biaxially along longitudinal and transverse axes of the beating whole heart. To determine whether neuropeptides that are known to modulate cardiac activity in ITALIC! H. americanusaffect the active or passive components of these length-tension curves, we also performed these tests in the presence of SGRNFLRFamide (SGRN) and GYSNRNYLRFamide (GYS). In uniaxial and biaxial tests, both passive and active forces increased with stretch along both measurement axes. The increase in passive forces was anisotropic, with greater increases along the longitudinal axis. Passive forces showed hysteresis and active forces were higher during lengthening than shortening phases of the stretch pyramid. Active forces at a given length were increased by both neuropeptides. To exert these effects, neuropeptides might have acted indirectly on the muscle via their effects on the cardiac ganglion, directly on the neuromuscular junction, or directly on the muscles. Because increases in response to stretch were also seen in stimulated motor nerve-muscle preparations, at least some of the effects of the peptides are likely peripheral. Taken together, these findings suggest that flexibility in rhythmic cardiac contractions results from the amplified effects of neuropeptides interacting with the length-tension characteristics of the heart. PMID:26896540

  8. Toxicity and QSAR of chlorobenzenes in two species of Benthic flatfish, flounder (Platichthys flesus L.) and sole (Solea solea L.)

    SciTech Connect

    Furay, V.J.; Smith, S.

    1995-01-01

    Chlorinated benzenes were among the first large-scale produced aromatic compounds. They have found broad spectrum of uses in numerous domestic and industrial preparations, ranging from engine - block cleaners, solvents, pharmaceutical intermediates, synthesis of chlorophenols and in disinfectants. Recent investigations have shown that they are present in the major environmental compartments and organisms, including fish tissues, coastal waters, and estuaries. Comparatively little is known about the ecotoxicity of chlorinated benzenes particularly to economically important fish. A number of earlier investigations examined the toxicity of chlorobenzenes and they demonstrated differences in the toxicity values for the same species and isomers of the chemicals investigated. The present investigation has assessed the toxicity and QSARs of selected chlorobenzenes to two ecologically and commercially important flatfish, the flounder (platichthys flesus) and sole (Solea solea). Both have a widespread geographic distribution in coastal and estuarine regions throughout Western Europe and are therefore highly representative test species. 20 refs., 5 tabs.

  9. Antimicrobial activity of peptides derived from olive flounder lipopolysaccharide binding protein/bactericidal permeability-increasing protein (LBP/BPI).

    PubMed

    Nam, Bo-Hye; Moon, Ji-Young; Park, Eun-Hee; Kim, Young-Ok; Kim, Dong-Gyun; Kong, Hee Jeong; Kim, Woo-Jin; Jee, Young Ju; An, Cheul Min; Park, Nam Gyu; Seo, Jung-Kil

    2014-10-17

    We describe the antimicrobial function of peptides derived from the C-terminus of the olive flounder LBP BPI precursor protein. The investigated peptides, namely, ofLBP1N, ofLBP2A, ofLBP4N, ofLBP5A, and ofLBP6A, formed α-helical structures, showing significant antimicrobial activity against several Gram-negative bacteria, Gram-positive bacteria, and the yeast Candida albicans, but very limited hemolytic activities. The biological activities of these five analogs were evaluated against biomembranes or artificial membranes for the development of candidate therapeutic agents. Gel retardation studies revealed that peptides bound to DNA and inhibited migration on an agarose gel. In addition, we demonstrated that ofLBP6A inhibited polymerase chain reaction. These results suggested that the ofLBP-derived peptide bactericidal mechanism may be related to the interaction with intracellular components such as DNA or polymerase.

  10. Antimicrobial Activity of Peptides Derived from Olive Flounder Lipopolysaccharide Binding Protein/Bactericidal Permeability-Increasing Protein (LBP/BPI)

    PubMed Central

    Nam, Bo-Hye; Moon, Ji-Young; Park, Eun-Hee; Kim, Young-Ok; Kim, Dong-Gyun; Kong, Hee Jeong; Kim, Woo-Jin; Jee, Young Ju; An, Cheul Min; Park, Nam Gyu; Seo, Jung-Kil

    2014-01-01

    We describe the antimicrobial function of peptides derived from the C-terminus of the olive flounder LBP BPI precursor protein. The investigated peptides, namely, ofLBP1N, ofLBP2A, ofLBP4N, ofLBP5A, and ofLBP6A, formed α-helical structures, showing significant antimicrobial activity against several Gram-negative bacteria, Gram-positive bacteria, and the yeast Candida albicans, but very limited hemolytic activities. The biological activities of these five analogs were evaluated against biomembranes or artificial membranes for the development of candidate therapeutic agents. Gel retardation studies revealed that peptides bound to DNA and inhibited migration on an agarose gel. In addition, we demonstrated that ofLBP6A inhibited polymerase chain reaction. These results suggested that the ofLBP-derived peptide bactericidal mechanism may be related to the interaction with intracellular components such as DNA or polymerase. PMID:25329706

  11. Distribution and expression in vitro and in vivo of DNA vaccine against lymphocystis disease virus in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Zheng, Fengrong; Sun, Xiuqin; Liu, Hongzhan; Wu, Xingan; Zhong, Nan; Wang, Bo; Zhou, Guodong

    2010-01-01

    Lymphocystis disease, caused by the lymphocystis disease virus (LCDV), is a significant worldwide problem in fish industry causing substantial economic losses. In this study, we aimed to develop the DNA vaccine against LCDV, using DNA vaccination technology. We evaluated plasmid pEGFP-N2-LCDV1.3 kb as a DNA vaccine candidate. The plasmid DNA was transiently expressed after liposome transfection into the eukaryotic COS 7 cell line. The distribution and expression of the DNA vaccine (pEGFP-N2-LCDV1.3kb) were also analyzed in tissues of the vaccinated Japanese flounder by PCR, RT-PCR and fluorescent microscopy. Results from PCR analysis indicated that the vaccine-containing plasmids were distributed in injected muscle, the muscle opposite the injection site, the hind intestine, gill, spleen, head, kidney and liver, 6 and 25 days after vaccination. The vaccine plasmids disappeared 100 d post-vaccination. Fluorescent microscopy revealed green fluorescence in the injected muscle, the muscle opposite the injection site, the hind intestine, gill, spleen, head, kidney and liver of fish 48 h post-vaccination, green fluorescence did not appear in the control treated tissue. Green fluorescence became weak at 60 days post-vaccination. RT-PCR analysis indicated that the mcp gene was expressed in all tested tissues of vaccinated fish 6-50 days post-vaccination. These results demonstrate that the antigen encoded by the DNA vaccine is distributed and expressed in all of the tissues analyzed in the vaccinated fish. The antigen would therefore potentially initiate a specific immune response. the plasmid DNA was injected into Japanese flounder ( Paralichthys olivaceus) intramuscularly and antibodies against LCDV were evaluated. The results indicate that the plasmid encoded DNA vaccine could induce an immune response to LCDV and would therefore offer immune protection against LCD. Further studies are required for the development and application of this promising DNA vaccine.

  12. Quantitative trait loci detection of Edwardsiella tarda resistance in Japanese flounder Paralichthys olivaceus using bulked segregant analysis

    NASA Astrophysics Data System (ADS)

    Wang, Xiaoxia; Xu, Wenteng; Liu, Yang; Wang, Lei; Sun, Hejun; Wang, Lei; Chen, Songlin

    2016-11-01

    In recent years, Edwardsiella tarda has become one of the most deadly pathogens of Japanese flounder ( Paralichthys olivaceus), causing serious annual losses in commercial production. In contrast to the rapid advances in the aquaculture of P. olivaceus, the study of E. tarda resistance-related markers has lagged behind, hindering the development of a disease-resistant strain. Thus, a marker-trait association analysis was initiated, combining bulked segregant analysis (BSA) and quantitative trait loci (QTL) mapping. Based on 180 microsatellite loci across all chromosomes, 106 individuals from the F1333 (♀: F0768 ×♂: F0915) (Nomenclature rule: F+year+family number) were used to detect simple sequence repeats (SSRs) and QTLs associated with E. tarda resistance. After a genomic scan, three markers (Scaffold 404-21589, Scaffold 404-21594 and Scaffold 270-13812) from the same linkage group (LG)-1 exhibited a significant difference between DNA, pooled/bulked from the resistant and susceptible groups (P <0.001). Therefore, 106 individuals were genotyped using all the SSR markers in LG1 by single marker analysis. Two different analytical models were then employed to detect SSR markers with different levels of significance in LG1, where 17 and 18 SSR markers were identified, respectively. Each model found three resistance-related QTLs by composite interval mapping (CIM). These six QTLs, designated qE1-6, explained 16.0%-89.5% of the phenotypic variance. Two of the QTLs, qE-2 and qE-4, were located at the 66.7 cM region, which was considered a major candidate region for E. tarda resistance. This study will provide valuable data for further investigations of E. tarda resistance genes and facilitate the selective breeding of disease-resistant Japanese flounder in the future.

  13. Quantitative trait loci detection of Edwardsiella tarda resistance in Japanese flounder Paralichthys olivaceus using bulked segregant analysis

    NASA Astrophysics Data System (ADS)

    Wang, Xiaoxia; Xu, Wenteng; Liu, Yang; Wang, Lei; Sun, Hejun; Wang, Lei; Chen, Songlin

    2016-03-01

    In recent years, Edwardsiella tarda has become one of the most deadly pathogens of Japanese flounder (Paralichthys olivaceus), causing serious annual losses in commercial production. In contrast to the rapid advances in the aquaculture of P. olivaceus, the study of E. tarda resistance-related markers has lagged behind, hindering the development of a disease-resistant strain. Thus, a marker-trait association analysis was initiated, combining bulked segregant analysis (BSA) and quantitative trait loci (QTL) mapping. Based on 180 microsatellite loci across all chromosomes, 106 individuals from the F1333 (♀: F0768 ×♂: F0915) (Nomenclature rule: F+year+family number) were used to detect simple sequence repeats (SSRs) and QTLs associated with E. tarda resistance. After a genomic scan, three markers (Scaffold 404-21589, Scaffold 404-21594 and Scaffold 270-13812) from the same linkage group (LG)-1 exhibited a significant difference between DNA, pooled/bulked from the resistant and susceptible groups (P <0.001). Therefore, 106 individuals were genotyped using all the SSR markers in LG1 by single marker analysis. Two different analytical models were then employed to detect SSR markers with different levels of significance in LG1, where 17 and 18 SSR markers were identified, respectively. Each model found three resistance-related QTLs by composite interval mapping (CIM). These six QTLs, designated qE1-6, explained 16.0%-89.5% of the phenotypic variance. Two of the QTLs, qE-2 and qE-4, were located at the 66.7 cM region, which was considered a major candidate region for E. tarda resistance. This study will provide valuable data for further investigations of E. tarda resistance genes and facilitate the selective breeding of disease-resistant Japanese flounder in the future.

  14. CpG-ODN increases resistance of olive flounder (Paralichthys olivaceus) against Philasterides dicentrarchi (Ciliophora: Scuticociliatia) infection.

    PubMed

    Lee, Eun Hye; Kim, Ki Hong

    2009-01-01

    Unmethylated cytosine-phosphate-guanine (CpG) dinucleotides flanked by specific bases in bacterial DNA induce a favorable immune response by acting as danger signals to the host. Synthetic oligodeoxynucleotides containing CpG motifs (CpG-ODNs) also act like the unmethylated CpG oligonucleotides in bacterial DNA. In the present study, we investigated the effects of synthetic CpG-ODN on the protection of olive flounder (Paralichthys olivaceus) against infection by Philasterides dicentrarchi, a pathogen of scuticociliatosis, through two consecutive experiments (trial I and II). Fish were intraperitoneally (i.p.) injected with CpG-ODN 1668 or GpC-ODN 1720 at different doses (3 microg in trial I and 10 microg in trial II), and after one week the fish were i.p. challenged with P. dicentrarchi. In both trial I and II, fish injected with CpG-ODN 1668 showed significantly higher serum scuticocidal activity than fish injected with PBS alone, while the scuticocidal activity disappeared by heat-inactivation. This result suggests that CpG-ODN might activate an alternative pathway of complement of olive flounder, and complement-mediated killing might be an important innate immune factor in the resistance against P. dicentrarchi infection. Although the cumulative mortality was largely different between trials I and II, the relative survival rate of fish injected with a high dose of CpG-ODN 1668 was considerably higher than that of fish injected with a low dose of this ODN, while the relative survival rate was not different between fish injected with the high dose and low dose of GpC-ODN 1720. The results of the present study suggest that CpG-ODNs may be used as potential immunostimulants to lessen cultured fish loss caused by scuticociliates. PMID:18992348

  15. Molecular cloning, mRNA expression and enzymatic characterization of cathepsin F from olive flounder (Paralichthys olivaceus).

    PubMed

    Ahn, Sang Jung; Kim, Na Young; Seo, Jung Soo; Je, Ju Eun; Sung, Ji Hea; Lee, Sang Hwan; Kim, Moo-Sang; Kim, Joong Kyun; Chung, Joon Ki; Lee, Hyung Ho

    2009-10-01

    Cathepsin F is a recently described papain-like cysteine protease of unknown function, and unique among cathepsins due to an elongated N-terminal pro-region, which contains a cystatin domain. In the present study, the cDNA of olive flounder (Paralichthys olivaceus) cathepsin F (PoCtF) was cloned by the combination of homology molecular cloning and rapid amplification of cDNA ends (RACE) approaches. The PoCtF gene was determined to consist of the 1844 bp nucleotide sequence which encodes for a 475-amino acid polypeptide. The results of RT-PCR analysis revealed ubiquitous expression throughout the entirety of healthy flounder tissues; however the PoCtF expressions increased significantly in gill at 3h post-injection with lipopolysaccharide (LPS). Also, immunostaining using anti-PoCtF antibody was strongest on the epidermal mucus in the fin. The cDNA encoding mature enzyme of PoCtF was expressed in Escherichia coli using the pGEX-4T-1 expression vector system. Its activity was quantified by cleaving the synthetic peptide Z-Phe-Arg-AMC, a substrate commonly used for functional characterization of cysteine proteinases, and the optimal pH for the protease activity was 7.5. The findings of the present study suggest that PoCtF has a higher optimum pH than mammalian cathepsin F, and PoCtF is an interesting target for future investigations of the role of cathepsin F in the epidermal mucus and fish innate immune system. PMID:19545641

  16. Molecular cloning, expression pattern, and 3D structural prediction of the cold inducible RNA-binding protein (CIRP) in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Yang, Xiao; Gao, Jinning; Ma, Liman; Li, Zan; Wang, Wenji; Wang, Zhongkai; Yu, Haiyang; Qi, Jie; Wang, Xubo; Wang, Zhigang; Zhang, Quanqi

    2015-02-01

    Cold-inducible RNA-binding protein (CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first cloned from mammals. On the contrary, there are little reports in teleosts. In this study, the Po CIRP gene of the Japanese flounder was cloned and sequenced. The genomic sequence consists of seven exons and six introns. The putative PoCIRP protein of flounder was 198 amino acid residues long containing the RNA recognition motif (RRM). Phylogenetic analysis showed that the flounder PoCIRP is highly conserved with other teleost CIRPs. The 5' flanking sequence was cloned by genome walking and many transcription factor binding sites were identified. There is a CpGs region located in promoter and exon I region and the methylation state is low. Quantitative real-time PCR analysis uncovered that Po CIRP gene was widely expressed in adult tissues with the highest expression level in the ovary. The mRNA of the Po CIRP was maternally deposited and the expression level of the gene was regulated up during the gastrula and neurula stages. In order to gain the information how the protein interacts with mRNA, we performed the modeling of the 3D structure of the flounder PoCIRP. The results showed a cleft existing the surface of the molecular. Taken together, the results indicate that the CIRP is a multifunctional molecular in teleosts and the findings about the structure provide valuable information for understanding the basis of this protein's function.

  17. The importance of rivers as nursery grounds for 0- and 1-group flounder ( Platichthys flesus L.) in comparison to the Wadden sea

    NASA Astrophysics Data System (ADS)

    Kerstan, M.

    From May 1988 to June 1989 selected areas of the German Wadden Sea, the Ems, Weser, Elbe and Eider estuaries as well as in Elbe tributaries were sampled for 0- and 1-group flounder ( Platichthys flesus L.). Estuarine and Wadden Sea sampling was conducted on board commercial shrimp vessels equipped with beam trawls. The Elbe river system was sampled on board a research boat equipped with a similar 3-m beam trawl. Density indices were calculated for the various areas. To estimate the importance of rivers as nursery areas for flounder, river surface area was compared to the area of tidal flats in the Wadden Sea. In all surveys, there was no relationship between fish densities and tow directions with respect to tide. In the Elbe river system flounder densities were not correlated to substrate types but increased significantly with decreasing salinity. Abundances were always lowestt in polyhaline habitats and increased up to 10-fold in mesohaline estuarine areas. In the limnetic tidal sections of the Elbe river and its tributaries densities increased again by factors of 3 to 10. The succession of 1-group modal lengths from limnetic to polyhaline habitats demonstrated that smaller fish preferred less saline waters. 0-group specimens lagging behind in growth did not leave the limnetic river section in winter. The onset of the spawning migration in November was determined by monitoring the length-frequency distributions throughout the year. The possible sources of bias are discussed and gear efficiency is estimated from literature sources. The contribution of rivers to the 0- and 1-group flounder population on the tidal flats of the Wadden Sea is estimated at about 35.1%.

  18. A multiple endpoint analysis of the effects of chronic exposure to sediment contaminated with Deepwater Horizon oil on juvenile Southern flounder and their associated microbiomes.

    PubMed

    Brown-Peterson, Nancy J; Krasnec, Michelle; Takeshita, Ryan; Ryan, Caitlin N; Griffitt, Kimberly J; Lay, Claire; Mayer, Gregory D; Bayha, Keith M; Hawkins, William E; Lipton, Ian; Morris, Jeffrey; Griffitt, Robert J

    2015-08-01

    Exposure to oiled sediments can negatively impact the health of fish species. Here, we examine the effects of chronic exposure of juvenile southern flounder, Paralichthys lethostigma, to a sediment-oil mixture. Oil:sediment mixtures are persistent over time and can become bioavailable following sediment perturbation or resuspension. Juvenile flounder were exposed for 32 days under controlled laboratory conditions to five concentrations of naturally weathered Macondo MC252 oil mixed into uncontaminated, field-collected sediments. The percent composition of individual polycyclic aromatic hydrocarbons (PAHs) of the weathered oil did not change after mixing with the sediment. Spiked exposure sediments contained 0.04-395mg/kg tPAH50 (sum of 50 individual PAH concentration measurements). Mortality increased with both exposure duration and concentration of sediment-associated PAHs, and flounder exposed to concentrations above 8mg/kg tPAH50 showed significantly reduced growth over the course of the experiment. Evident histopathologic changes were observed in liver and gill tissues of fish exposed to more than 8mg/kg tPAH50. All fish at these concentrations showed hepatic intravascular congestion, macrovesicular hepatic vacoulation, telangiectasia of secondary lamellae, and lamellar epithelial proliferation in gill tissues. Dose-dependent upregulation of Cyp1a expression in liver tissues was observed. Taxonomic analysis of gill and intestinal commensal bacterial assemblages showed that exposure to oiled sediments led to distinct shifts in commensal bacterial population structures. These data show that chronic exposure to environmentally-relevant concentrations of oiled sediments produces adverse effects in flounder at multiple biological levels. PMID:26092636

  19. Mass spectral characterization of peptide transmitters/hormones in the nervous system and neuroendocrine organs of the American lobster Homarus americanus

    PubMed Central

    Ma, Mingming; Chen, Ruibing; Sousa, Gregory L.; Bors, Eleanor K.; Kwiatkowski, Molly; Goiney, Christopher C.; Goy, Michael F.; Christie, Andrew E.; Li, Lingjun

    2008-01-01

    The American lobster Homarus americanus is a decapod crustacean with both high economic and scientific importance. To facilitate physiological investigations of peptide transmitter/hormone function in this species, we have used matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and nanoscale liquid chromatography coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry (nanoLC-ESI-Q-TOF-MS/MS) to elucidate the peptidome present in its nervous system and neuroendocrine organs. In total, 84 peptides were identified, including 27 previously known H. americanus peptides (e.g. VYRKPPFNGSIFamide [Val1-SIFamide]), 23 peptides characterized previously from other decapods, but new to the American lobster (e.g. pQTFQYSRGWTNamide [Arg7-corazonin]), and 34 new peptides de novo sequenced/detected for the first time in this study. Of particular note are a novel B-type allatostatin (TNWNKFQGSWamide) and several novel FMRFamide-related peptides, including an unsulfated analog of sulfakinin (GGGEYDDYGHLRFamide), two myosuppressins (QDLDHVFLRFamide and pQDLDHVFLRFamide), and a collection of short neuropeptide F isoforms (e.g. DTSTPALRLRFamide, and FEPSLRLRFamide). Our data also include the first detection of multiple tachykinin-related peptides in a non-brachyuran decapod, as well as the identification of potential individual-specific variants of orcokinin and orcomyotropin-related peptide. Taken collectively, our results not only expand greatly the number of known H. americanus neuropeptides, but also provide a framework for future studies on the physiological roles played by these molecules in this commercially and scientifically important species. PMID:18304551

  20. Expression, purification, and characterization of the Necator americanus aspartic protease-1 (Na-APR-1 (M74)) antigen, a component of the bivalent human hookworm vaccine

    PubMed Central

    Seid, Christopher A; Curti, Elena; Jones, R Mark; Hudspeth, Elissa; Rezende, Wanderson; Pollet, Jeroen; Center, Lori; Versteeg, Leroy; Pritchard, Sonya; Musiychuk, Konstantin; Yusibov, Vidadi; Hotez, Peter J; Bottazzi, Maria Elena

    2015-01-01

    Over 400 million people living in the world's poorest developing nations are infected with hookworms, mostly of the genus Necator americanus. A bivalent human hookworm vaccine composed of the Necator americanus Glutathione S-Transferase-1 (Na-GST-1) and the Necator americanus Aspartic Protease-1 (Na-APR-1 (M74)) is currently under development by the Sabin Vaccine Institute Product Development Partnership (Sabin PDP). Both monovalent vaccines are currently in Phase 1 trials. Both Na-GST-1 and Na-APR-1 antigens are expressed as recombinant proteins. While Na-GST-1 was found to express with high yields in Pichia pastoris, the level of expression of Na-APR-1 in this host was too low to be suitable for a manufacturing process. When the tobacco plant Nicotiana benthamiana was evaluated as an expression system, acceptable levels of solubility, yield, and stability were attained. Observed expression levels of Na-APR-1 (M74) using this system are ∼300 mg/kg. Here we describe the achievements and obstacles encountered during process development as well as characterization and stability of the purified Na-APR-1 (M74) protein and formulated vaccine. The expression, purification and analysis of purified Na-APR-1 (M74) protein obtained from representative 5 kg reproducibility runs performed to qualify the Na-APR-1 (M74) production process is also presented. This process has been successfully transferred to a pilot plant and a 50 kg scale manufacturing campaign under current Good Manufacturing Practice (cGMP) has been performed. The 50 kg run has provided a sufficient amount of protein to support the ongoing hookworm vaccine development program of the Sabin PDP. PMID:25905574

  1. Sex, Diet, and the Social Environment: Factors Influencing Hair Cortisol Concentration in Free-Ranging Black Bears (Ursus americanus).

    PubMed

    Lafferty, Diana J R; Laudenslager, Mark L; Mowat, Garth; Heard, Doug; Belant, Jerrold L

    2015-01-01

    Increasingly, measures of glucocorticoid levels (e.g., cortisol), key components of the neuroendocrine stress axis, are being used to measure past hypothalamic-pituitary-adrenal (HPA) activity to index psychological and physiological stress exhibited by wildlife for assessing individual and population-level well-being. However, many intrinsic and extrinsic factors affect HPA activity in animals. Using American black bears (Ursus americanus; n = 116) as an ecological model and hair cortisol concentration (HCC) as an integrative measure of past HPA activity, we evaluated the influence of diet, sex and the social environment on black bear HCC in a free-ranging population that spanned adjoining ecoregions with differing densities of potential conspecific and heterospecific competitors. HCC varied by sex, with female HCC ranging from 0.6 to 10.7 pg/mg (median = 4.5 ± 1.2 mean absolute deviation [MAD]) and male HCC ranging from 0.5 to 35.1 pg/mg (median = 6.2 ± 2.6 MAD). We also observed a three-way interaction among sex, δ14C and ecoregion, which may indicate that some differences in HCC between female and male black bears results from variability in the nutritional needs of larger-bodied males relative to smaller-bodied females, slight differences in food resources use between ecoregions as well as sex-based differences regarding the social environment. Once we understand what drives sex-specific differences in HCC, HCC may aid our understanding of the physiological responses by bears and other wildlife to diverse environmental challenges.

  2. Wound healing during hibernation by black bears (Ursus americanus) in the wild: elicitation of reduced scar formation.

    PubMed

    Iaizzo, Paul A; Laske, Timothy G; Harlow, Henry J; McClay, Carolyn B; Garshelis, David L

    2012-03-01

    Even mildly hypothermic body or limb temperatures can retard healing processes in mammals. Despite this, we observed that hibernating American black bears (Ursus americanus Pallas, 1780) elicit profound abilities in mounting inflammatory responses to infection and/or foreign bodies. In addition, they resolve injuries during hibernation while maintaining mildly hypothermic states (30-35 °C) and without eating, drinking, urinating or defecating. We describe experimental studies on free-ranging bears that document their abilities to completely resolve cutaneous cuts and punctures incurred during or prior to hibernation. We induced small, full-thickness cutaneous wounds (biopsies or incisions) during early denning, and re-biopsied sites 2-3 months later (near the end of denning). Routine histological methods were used to characterize these skin samples. All biopsied sites with respect to secondary intention (open circular biopsies) and primary intention (sutured sites) healed, with evidence of initial eschar (scab) formation, completeness of healed epidermis and dermal layers, dyskeratosis (inclusion cysts), and abilities to produce hair follicles. These healing abilities of hibernating black bears are a clear survival advantage to animals injured before or during denning. Bears are known to have elevated levels of hibernation induction trigger (delta-opioid receptor agonist) and ursodeoxycholic acid (major bile acid within plasma, mostly conjugated with taurine) during hibernation, which may relate to these wound-healing abilities. Further research as to the underlying mechanisms of wound healing during hibernation could have applications in human medicine. Unique approaches may be found to improve healing for malnourished, hypothermic, diabetic and elderly patients or to reduce scarring associated with burns and traumatic injuries.

  3. Sex, Diet, and the Social Environment: Factors Influencing Hair Cortisol Concentration in Free-Ranging Black Bears (Ursus americanus).

    PubMed

    Lafferty, Diana J R; Laudenslager, Mark L; Mowat, Garth; Heard, Doug; Belant, Jerrold L

    2015-01-01

    Increasingly, measures of glucocorticoid levels (e.g., cortisol), key components of the neuroendocrine stress axis, are being used to measure past hypothalamic-pituitary-adrenal (HPA) activity to index psychological and physiological stress exhibited by wildlife for assessing individual and population-level well-being. However, many intrinsic and extrinsic factors affect HPA activity in animals. Using American black bears (Ursus americanus; n = 116) as an ecological model and hair cortisol concentration (HCC) as an integrative measure of past HPA activity, we evaluated the influence of diet, sex and the social environment on black bear HCC in a free-ranging population that spanned adjoining ecoregions with differing densities of potential conspecific and heterospecific competitors. HCC varied by sex, with female HCC ranging from 0.6 to 10.7 pg/mg (median = 4.5 ± 1.2 mean absolute deviation [MAD]) and male HCC ranging from 0.5 to 35.1 pg/mg (median = 6.2 ± 2.6 MAD). We also observed a three-way interaction among sex, δ14C and ecoregion, which may indicate that some differences in HCC between female and male black bears results from variability in the nutritional needs of larger-bodied males relative to smaller-bodied females, slight differences in food resources use between ecoregions as well as sex-based differences regarding the social environment. Once we understand what drives sex-specific differences in HCC, HCC may aid our understanding of the physiological responses by bears and other wildlife to diverse environmental challenges. PMID:26529405

  4. Localization and characterization of neuropeptide Y-like peptides in the brain and islet organ of the anglerfish (Lophius americanus).

    PubMed

    Noe, B D; Milgram, S L; Balasubramaniam, A; Andrews, P C; Calka, J; McDonald, J K

    1989-08-01

    Results from a previous report demonstrate that more than one molecular form of neuropeptide Y-like peptide may be present in the islet organ of the anglerfish (Lophius americanus). Most of the neuropeptide Y-like immunoreactive material was anglerfish peptide YG, which is expressed in a subset of islet cells, whereas an additional neuropeptide Y-like peptide(s) was localized in islet nerves. To learn more about the neuropeptide Y-like peptides in islet nerves, we have employed immunohistochemical and biochemical methods to compare peptides found in anglerfish islets and brain. Using antisera that selectively react with either mammalian forms of neuropeptide Y or with anglerfish peptide YG, subsets of neurons were found in the brain that labelled with only one or the other of the antisera. In separate sections, other neurons that were labelled with either antiserum exhibited similar morphologies. Peptides from brains and islets were subjected to gel filtration and reverse-phase high performance liquid chromatography. Radioimmunoassays employing either the neuropeptide Y or peptide YG antisera were used to examine chromatographic eluates. Immunoreactive peptides having retention times of human neuropeptide Y and porcine neuropeptide Y were identified in extracts of both brain and islets. This indicates that peptides structurally similar to both of these peptides from the neuropeptide Y-pancreatic polypeptide family are expressed in neurons of anglerfish brain and nerve fibers of anglerfish islets. The predominant form of neuropeptide Y-like peptide in islets was anglerfish peptide YG. Neuropeptide Y-immunoreactive peptides from islet extracts that had chromatographic retention times identical to human neuropeptide Y and porcine neuropeptide Y were present in much smaller quantities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2673525

  5. (Lack of) genetic diversity in immune genes predates glacial isolation in the North American mountain goat (Oreamnos americanus).

    PubMed

    Shafer, Aaron B A; Fan, Chia Wei; Côté, Steeve D; Coltman, David W

    2012-01-01

    The major histocompatibility complex (MHC) plays an important role in an organism's ability to respond to pathogens. Immunogenetic diversity is advantageous as it permits the recognition of more external antigens. For this reason, MHC and immune gene variation are considered a barometer for the genetic health of wild populations. Mountain goats (Oreamnos americanus) were previously shown to have little variation at the MHC Class II Oram-DRB locus, which was attributed to population bottlenecks during the last glacial maximum (LGM). In this paper, we extended the analysis of immunogenetic variability in mountain goats to 5 genes representing the 3 classes of MHC gene (Class I OLA, Class II DRA and DRB, and Class III TNF-α) and the natural resistance-associated macrophage protein. We sequenced approximately 3000 bp from 31 individuals sampled across the range of mountain goats and found very low levels of diversity (1-3 polymorphic sites per gene) with the exception of the Class I Oram-OLA gene. Oram-OLA was nearly 30 times more diverse than the other immune genes and appears to represent a source of increased immunogenetic diversity. This diversity may be attributed to multiple loci, mediated by pathogen exposure, or potentially influenced by social factors. The distribution of SNPs was not associated with refugial history, suggesting that the current distribution of immunogenetic diversity was present prior to the LGM. These data suggest that although they have low levels of diversity at the 4 of 5 immune loci, mountain goats may be better equipped for future climate oscillations and pathogen exposure than previously thought. PMID:22268162

  6. Kinematics, hydrodynamics and force production of pleopods suggest jet-assisted walking in the American lobster (Homarus americanus).

    PubMed

    Lim, Jeanette L; Demont, M Edwin

    2009-09-01

    The American lobster (Homarus americanus) displays a diverse set of locomotory behaviours that includes tail flips, walking and paddling. Paddling is carried out by the four pairs of paddle-shaped pleopods on the ventral abdomen. Although it is recognized that pleopod-generated fluid flows have some locomotory role in adults, reports on their relative importance in locomotion are inconsistent. This paper integrates experimental kinematics and hydrodynamics of lobster pleopod beating to determine the mechanism and magnitude of pleopod force production. A kinematic analysis of pleopod beating in live lobsters showed that the pleopods execute an adlocomotory metachronal beating pattern. We modelled in vivo pleopod kinematics with a set of simple trigonometric functions, and used these functions to program a mechanical lobster model consisting of motor-driven pleopods on a lobster abdomen exoskeleton. Based on flow visualizations obtained from applying particle image velocimetry to the lobster model, we propose that the unsteady metachronal kinematics of the pleopods can maximize thrust by exploiting forces arising from individual pleopod activity and interactions among adjacent pairs. The pleopods continuously entrain fluid surrounding the lobster and create a caudally directed fluid jet oriented parallel to the substratum. Inputting wake morphology and velocity data into a simplified model for steady jet thrust showed that the pleopods of the lobster model produced 27-54 mN of thrust, which is comparable to the propulsive forces generated by other proficient swimmers. These results suggest that lobster pleopods are capable of producing forces of a magnitude that could assist the walking legs in forward propulsion.

  7. Wound healing during hibernation by black bears (Ursus americanus) in the wild: elicitation of reduced scar formation.

    PubMed

    Iaizzo, Paul A; Laske, Timothy G; Harlow, Henry J; McClay, Carolyn B; Garshelis, David L

    2012-03-01

    Even mildly hypothermic body or limb temperatures can retard healing processes in mammals. Despite this, we observed that hibernating American black bears (Ursus americanus Pallas, 1780) elicit profound abilities in mounting inflammatory responses to infection and/or foreign bodies. In addition, they resolve injuries during hibernation while maintaining mildly hypothermic states (30-35 °C) and without eating, drinking, urinating or defecating. We describe experimental studies on free-ranging bears that document their abilities to completely resolve cutaneous cuts and punctures incurred during or prior to hibernation. We induced small, full-thickness cutaneous wounds (biopsies or incisions) during early denning, and re-biopsied sites 2-3 months later (near the end of denning). Routine histological methods were used to characterize these skin samples. All biopsied sites with respect to secondary intention (open circular biopsies) and primary intention (sutured sites) healed, with evidence of initial eschar (scab) formation, completeness of healed epidermis and dermal layers, dyskeratosis (inclusion cysts), and abilities to produce hair follicles. These healing abilities of hibernating black bears are a clear survival advantage to animals injured before or during denning. Bears are known to have elevated levels of hibernation induction trigger (delta-opioid receptor agonist) and ursodeoxycholic acid (major bile acid within plasma, mostly conjugated with taurine) during hibernation, which may relate to these wound-healing abilities. Further research as to the underlying mechanisms of wound healing during hibernation could have applications in human medicine. Unique approaches may be found to improve healing for malnourished, hypothermic, diabetic and elderly patients or to reduce scarring associated with burns and traumatic injuries. PMID:22405448

  8. Copper-metallothioneins in the American lobster, Homarus americanus: potential role as Cu(I) donors to apohemocyanin.

    PubMed Central

    Brouwer, M; Whaling, P; Engel, D W

    1986-01-01

    The physiological function of copper(I)-metallothionein is not well understood. The respiratory function of hemocyanin, a copper(I)-containing respiratory protein found in the hemolymph of many invertebrates, has been known a long time. However, the mechanism by which Cu(I) is inserted into the oxygen-binding site of apohemocyanin is completely unknown. This investigation tests the hypothesis that copper(I)-metallothionein may act as a Cu(I) donor to apohemocyanin. To this end, copper-binding proteins and hemocyanin were purified from the digestive gland and hemolymph of the American lobster, Homarus americanus. In the presence of beta-mercaptoethanol, the copper-binding proteins can be resolved into three components on DEAE-cellulose. The first two have been characterized as metallothioneins, based on their high cysteine content and lack of aromatic amino acid residues. The cysteine content of the third component is half of that of components I and II. In the absence of beta-mercaptoethanol the three proteins elute as a single protein complex during ion-exchange chromatography. Components I and II show a strong tendency to polymerize, a process that is accompanied by the loss of protein-bound copper. The purified proteins are not capable of transferring Cu(I) to the active sites of completely copper-free apohemocyanin. They are capable, however, of transferring Cu(I) to active sites of hemocyanin containing reduced amounts of Cu(I), suggesting that the conformational state of hemocyanin is the determining factor in the Cu(I) transfer mechanism. PMID:3709470

  9. (Lack of) genetic diversity in immune genes predates glacial isolation in the North American mountain goat (Oreamnos americanus).

    PubMed

    Shafer, Aaron B A; Fan, Chia Wei; Côté, Steeve D; Coltman, David W

    2012-01-01

    The major histocompatibility complex (MHC) plays an important role in an organism's ability to respond to pathogens. Immunogenetic diversity is advantageous as it permits the recognition of more external antigens. For this reason, MHC and immune gene variation are considered a barometer for the genetic health of wild populations. Mountain goats (Oreamnos americanus) were previously shown to have little variation at the MHC Class II Oram-DRB locus, which was attributed to population bottlenecks during the last glacial maximum (LGM). In this paper, we extended the analysis of immunogenetic variability in mountain goats to 5 genes representing the 3 classes of MHC gene (Class I OLA, Class II DRA and DRB, and Class III TNF-α) and the natural resistance-associated macrophage protein. We sequenced approximately 3000 bp from 31 individuals sampled across the range of mountain goats and found very low levels of diversity (1-3 polymorphic sites per gene) with the exception of the Class I Oram-OLA gene. Oram-OLA was nearly 30 times more diverse than the other immune genes and appears to represent a source of increased immunogenetic diversity. This diversity may be attributed to multiple loci, mediated by pathogen exposure, or potentially influenced by social factors. The distribution of SNPs was not associated with refugial history, suggesting that the current distribution of immunogenetic diversity was present prior to the LGM. These data suggest that although they have low levels of diversity at the 4 of 5 immune loci, mountain goats may be better equipped for future climate oscillations and pathogen exposure than previously thought.

  10. Sex, Diet, and the Social Environment: Factors Influencing Hair Cortisol Concentration in Free-Ranging Black Bears (Ursus americanus)

    PubMed Central

    Lafferty, Diana J. R.; Laudenslager, Mark L.; Mowat, Garth; Heard, Doug; Belant, Jerrold L.

    2015-01-01

    Increasingly, measures of glucocorticoid levels (e.g., cortisol), key components of the neuroendocrine stress axis, are being used to measure past hypothalamic-pituitary-adrenal (HPA) activity to index psychological and physiological stress exhibited by wildlife for assessing individual and population-level well-being. However, many intrinsic and extrinsic factors affect HPA activity in animals. Using American black bears (Ursus americanus; n = 116) as an ecological model and hair cortisol concentration (HCC) as an integrative measure of past HPA activity, we evaluated the influence of diet, sex and the social environment on black bear HCC in a free-ranging population that spanned adjoining ecoregions with differing densities of potential conspecific and heterospecific competitors. HCC varied by sex, with female HCC ranging from 0.6 to 10.7 pg/mg (median = 4.5 ± 1.2 mean absolute deviation [MAD]) and male HCC ranging from 0.5 to 35.1 pg/mg (median = 6.2 ± 2.6 MAD). We also observed a three-way interaction among sex, δ14C and ecoregion, which may indicate that some differences in HCC between female and male black bears results from variability in the nutritional needs of larger-bodied males relative to smaller-bodied females, slight differences in food resources use between ecoregions as well as sex-based differences regarding the social environment. Once we understand what drives sex-specific differences in HCC, HCC may aid our understanding of the physiological responses by bears and other wildlife to diverse environmental challenges. PMID:26529405

  11. Ink from longfin inshore squid, Doryteuthis pealeii, as a chemical and visual defense against two predatory fishes, summer flounder, Paralichthys dentatus, and sea catfish, Ariopsis felis.

    PubMed

    Derby, Charles D; Tottempudi, Mihika; Love-Chezem, Tiffany; Wolfe, Lanna S

    2013-12-01

    Chemical and visual defenses are used by many organisms to avoid being approached or eaten by predators. An example is inking molluscs-including gastropods such as sea hares and cephalopods such as squid, cuttlefish, and octopus-which release a colored ink upon approach or attack. Previous work showed that ink can protect molluscs through a combination of chemical, visual, and other effects. In this study, we examined the effects of ink from longfin inshore squid, Doryteuthis pealeii, on the behavior of two species of predatory fishes, summer flounder, Paralichthys dentatus, and sea catfish, Ariopsis felis. Using a cloud assay, we found that ink from longfin inshore squid affected the approach phase of predation by summer flounder, primarily through its visual effects. Using a food assay, we found that the ink affected the consummatory and ingestive phase of predation of both sea catfish and summer flounder, through the ink's chemical properties. Fractionation of ink showed that most of its deterrent chemical activity is associated with melanin granules, suggesting that either compounds adhering to these granules or melanin itself are the most biologically active. This work provides the basis for a comparative approach to identify deterrent molecules from inking cephalopods and to examine neural mechanisms whereby these chemicals affect behavior of fish, using the sea catfish as a chemosensory model.

  12. Antigen uptake and expression of antigen presentation-related immune genes in flounder (Paralichthys olivaceus) after vaccination with an inactivated Edwardsiella tarda immersion vaccine, following hyperosmotic treatment.

    PubMed

    Gao, Yingli; Tang, Xiaoqian; Sheng, Xiuzhen; Xing, Jing; Zhan, Wenbin

    2016-08-01

    Antigen uptake is a critical process for activation of the immune system, and therefore the ability to enhance antigen uptake is a primary consideration in the development of an immersion vaccination of fish. In the present work, flounders (Paralichthys olivaceus) were immersed in three hyperosmotic solutions with 40, 50 and 60‰ salinities, then transferred into seawater of normal salinity (i.e. 30‰) containing formalin-inactivated Edwardsiella tarda for 30 min. The antigen uptake in vaccinated flounder was determined using an absolute quantitative PCR (qPCR). The results showed significantly higher antigen uptake in the tissues of flounders immersed in solutions with 50‰ and 60‰ salinity compared to the control group directly immersed in vaccine (DI) (P < 0.05), and the highest amount of antigen was detected in flounders immersed in the 50‰ salinity solution, whereas there was no significant difference in antigen uptake between the 40‰ salinity group and the DI group (P > 0.05). A rapid and significant increase in antigen uptake was detected in the mucosal-associated tissues including the gill, skin and intestine (P < 0.05) compared with the spleen, kidney and liver. Antigen uptake in the gill and skin both peaked at 30 min post immersion, which was significantly higher than the levels of uptake measured in the other tissues (P < 0.05), and then quickly declined. In contrast, antigen uptake in the spleen, kidney and liver gradually increased 3 h post immersion (hpi). The expression profiles of four antigen presentation-related immune genes (MHC Iα, MHC IIα, CD4-1 and CD8α) were investigated after immersion. These four genes showed a significantly stronger response in the immersed flounders exposed to 50‰ salinity compared with the DI group (P < 0.05). In the mucosal-associated tissues, the expression of MHC Iα and CD8α genes peaked at 24 hpi, while the expression of MHC IIα and CD4-1 genes showed up-regulation in the gill and skin

  13. Post-Mating Interactions and Their Effects on Fitness of Female and Male Echinothrips americanus (Thysanoptera: Thripidae), a New Insect Pest in China

    PubMed Central

    Li, Xiao-Wei; Jiang, Hong-Xue; Zhang, Xiao-Chen; Shelton, Anthony M.; Feng, Ji-Nian

    2014-01-01

    Post-mating, sexual interactions of opposite sexes differ considerably in different organisms. Post-mating interactions such as re-mating behavior and male harassment can affect the fitness of both sexes. Echinothrips americanus is a new insect pest in Mainland China, and little is known about its post-mating interactions. In this study, we observed re-mating frequency and male harassment frequency and their effects on fitness parameters and offspring sex ratios of E. americanus females. Furthermore, we tested the impact of mating and post-mating interactions on fitness parameters of males. Our results revealed that the re-mating frequency in female adults was extremely low during a 30-day period. However, post-mating interactions between females and males, consisting mainly of male harassment and female resistance, did occur and significantly reduced female longevity and fecundity. Interestingly, increased access to males did not affect the ratio of female offspring. For males, mating dramatically reduced their longevity. However, post-mating interactions with females had no effects on the longevity of mated males. These results enrich our basic knowledge about female and male mating and post-mating behaviors in this species and provide important information about factors that may influence population regulation of this important pest species. PMID:24489956

  14. Investigating the mechanism for maintaining eucalcemia despite immobility and anuria in the hibernating American black bear (Ursus americanus).

    PubMed

    Seger, Rita L; Cross, Randal A; Rosen, Clifford J; Causey, Robert C; Gundberg, Caren M; Carpenter, Thomas O; Chen, Tai C; Halteman, William A; Holick, Michael F; Jakubas, Walter J; Keisler, Duane H; Seger, Richard M; Servello, Frederick A

    2011-12-01

    Ursine hibernation uniquely combines prolonged skeletal unloading, anuria, pregnancy, lactation, protein recycling, and lipolysis. This study presents a radiographic and biochemical picture of bone metabolism in free-ranging, female American black bears (Ursus americanus) that were active (spring bears and autumn bears) or hibernating (hibernating bears). Hibernating bears included lactating and non-lactating individuals. We measured serum calcium, albumin, inorganic phosphate, creatinine, bone specific alkaline phosphatase (BSALP), CTX, parathyroid hormone, insulin-like growth factor-I (IGF-l), leptin, 25-hydroxyvitamin D [25(OH)D], 1,25-dihydroxyvitamin D [1,25(OH)(2)D] and sclerostin from 35 to 50 tranquilized hibernating bears and 14 to 35 tranquilized spring bears. We compared metacarpal cortical indices (MCI), measured by digital X-ray radiogrammetry, from 60 hunter-killed autumn bears and 79 tranquilized, hibernating bears. MCI was greater in autumn than winter in younger bears, but showed no seasonal difference in older bears. During hibernation eucalcemia was maintained, BSALP was suppressed, and CTX was in the range expected for anuria. During hibernation 1,25(OH)(2)D was produced despite anuria. 1,25(OH)(2)D and IGF-I were less in hibernating than spring bears. In a quarter of hibernating bears, sclerostin was elevated. Leptin was greater in hibernating than spring bears. In hibernating bears, leptin correlated positively with BSALP in non-lactating bears and with CTX in lactating bears. Taken together the biochemical and radiographic findings indicate that during hibernation, bone turnover was persistent, balanced, and suppressed; bone resorption was lower than expected for an unloaded skeleton; and there was no unloading-induced bone loss. The skeleton appears to perceive that it was loaded when it was actually unloaded during hibernation. However, at the level of sclerostin, the skeleton recognized that it was unloaded. During hibernation leptin

  15. Genome-wide SNP identification for the construction of a high-resolution genetic map of Japanese flounder (Paralichthys olivaceus): applications to QTL mapping of Vibrio anguillarum disease resistance and comparative genomic analysis.

    PubMed

    Shao, Changwei; Niu, Yongchao; Rastas, Pasi; Liu, Yang; Xie, Zhiyuan; Li, Hengde; Wang, Lei; Jiang, Yong; Tai, Shuaishuai; Tian, Yongsheng; Sakamoto, Takashi; Chen, Songlin

    2015-04-01

    High-resolution genetic maps are essential for fine mapping of complex traits, genome assembly, and comparative genomic analysis. Single-nucleotide polymorphisms (SNPs) are the primary molecular markers used for genetic map construction. In this study, we identified 13,362 SNPs evenly distributed across the Japanese flounder (Paralichthys olivaceus) genome. Of these SNPs, 12,712 high-confidence SNPs were subjected to high-throughput genotyping and assigned to 24 consensus linkage groups (LGs). The total length of the genetic linkage map was 3,497.29 cM with an average distance of 0.47 cM between loci, thereby representing the densest genetic map currently reported for Japanese flounder. Nine positive quantitative trait loci (QTLs) forming two main clusters for Vibrio anguillarum disease resistance were detected. All QTLs could explain 5.1-8.38% of the total phenotypic variation. Synteny analysis of the QTL regions on the genome assembly revealed 12 immune-related genes, among them 4 genes strongly associated with V. anguillarum disease resistance. In addition, 246 genome assembly scaffolds with an average size of 21.79 Mb were anchored onto the LGs; these scaffolds, comprising 522.99 Mb, represented 95.78% of assembled genomic sequences. The mapped assembly scaffolds in Japanese flounder were used for genome synteny analyses against zebrafish (Danio rerio) and medaka (Oryzias latipes). Flounder and medaka were found to possess almost one-to-one synteny, whereas flounder and zebrafish exhibited a multi-syntenic correspondence. The newly developed high-resolution genetic map, which will facilitate QTL mapping, scaffold assembly, and genome synteny analysis of Japanese flounder, marks a milestone in the ongoing genome project for this species.

  16. Genome-wide SNP identification for the construction of a high-resolution genetic map of Japanese flounder (Paralichthys olivaceus): applications to QTL mapping of Vibrio anguillarum disease resistance and comparative genomic analysis.

    PubMed

    Shao, Changwei; Niu, Yongchao; Rastas, Pasi; Liu, Yang; Xie, Zhiyuan; Li, Hengde; Wang, Lei; Jiang, Yong; Tai, Shuaishuai; Tian, Yongsheng; Sakamoto, Takashi; Chen, Songlin

    2015-04-01

    High-resolution genetic maps are essential for fine mapping of complex traits, genome assembly, and comparative genomic analysis. Single-nucleotide polymorphisms (SNPs) are the primary molecular markers used for genetic map construction. In this study, we identified 13,362 SNPs evenly distributed across the Japanese flounder (Paralichthys olivaceus) genome. Of these SNPs, 12,712 high-confidence SNPs were subjected to high-throughput genotyping and assigned to 24 consensus linkage groups (LGs). The total length of the genetic linkage map was 3,497.29 cM with an average distance of 0.47 cM between loci, thereby representing the densest genetic map currently reported for Japanese flounder. Nine positive quantitative trait loci (QTLs) forming two main clusters for Vibrio anguillarum disease resistance were detected. All QTLs could explain 5.1-8.38% of the total phenotypic variation. Synteny analysis of the QTL regions on the genome assembly revealed 12 immune-related genes, among them 4 genes strongly associated with V. anguillarum disease resistance. In addition, 246 genome assembly scaffolds with an average size of 21.79 Mb were anchored onto the LGs; these scaffolds, comprising 522.99 Mb, represented 95.78% of assembled genomic sequences. The mapped assembly scaffolds in Japanese flounder were used for genome synteny analyses against zebrafish (Danio rerio) and medaka (Oryzias latipes). Flounder and medaka were found to possess almost one-to-one synteny, whereas flounder and zebrafish exhibited a multi-syntenic correspondence. The newly developed high-resolution genetic map, which will facilitate QTL mapping, scaffold assembly, and genome synteny analysis of Japanese flounder, marks a milestone in the ongoing genome project for this species. PMID:25762582

  17. Effects of phosphoglucomutase gene (PGM) in Streptococcus parauberis on innate immune response and pathogenicity of olive flounder (Paralichthys olivaceus).

    PubMed

    Woo, Sung Ho; Park, Soo Il

    2014-12-01

    In recent years, Streptococcus parauberis infection has been an emerging problem in aquaculture in South Korea because of its more frequent isolation than other streptococcal bacteria including Streptococcus iniae. To develop effective treatment and prophylaxis methods against this emerging disease by S. parauberis, it is necessary to understand the underlying pathogenic mechanisms. To uncover the pathogenicity, the mutant strain of S. parauberis with a deleted phosphoglucomutase (PGM) gene which has been known to be an important virulence factor in bacterial pathogens was generated to investigate the relationship between virulence and gene function using an allelic exchange mutagenesis method. Allelic exchange mutagenesis of the phosphoglucomutase gene resulted in phenotype changes including decreased extracellular capsules, reduced buoyancy, increased hydrophobicity and reduced growth. Moreover, the S. parauberis mutant was more sensitive to innate immune clearance mechanisms including serum, mucus and phagocyte killing and could not induce mortality in olive flounder. These phenotype changes and the attenuated virulence of the pathogen to fish could be due to the reduction in capsule production by mutation of the PGM gene. The results provide evidences that phosphoglucomutase expression contributes to S. parauberis virulence in fish by affecting bacterial survival against the host's humoral and cellular defense mechanisms.

  18. Establishment and characterization of a new marine fish cell line from ovary of barfin flounder ( Verasper moseri)

    NASA Astrophysics Data System (ADS)

    Xu, Xiaohui; Fan, Tingjun; Jiang, Guojian; Yang, Xiuxia

    2015-12-01

    A novel continuous ovary cell line from barfin flounder ( Verasper moseri) (BFO cell line) was established with its primitive application in transgenic expression demonstrated in this study. Primarily cultured cells grew well at 22°C in Dulbecco's modified Eagle medium/F12 medium (DMEM/F12, 1:1; pH 7.2) supplemented with 20% fetal bovine serum (FBS), carboxymethyl chitooligosaccharide, basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I). The primary BFO cells in fibroblastic morphology proliferated into a confluent monolayer about 2 weeks later, and were able to be subcultured. Impacts of medium and temperature on the growth of the cells were examined. The optimum growth was found in DMEM/F12 with 20% FBS and at 22°C. The BFO cells can be continuously subcultured to Passage 120 steadily with a population doubling time of 32.7 h at Passage 60. Chromosome analysis revealed that 72% of BFO cells at Passage 60 maintained the normal diploid chromosome number (46) with a normal karyotype of 2st+44t. The results of gene transformation indicated that green fluorescence protein (GFP) positively expressed in these cells after being transformed with pcDNA3.1-GFP. Therefore, a continuous and transformable BFO cell line was successfully established, which may serve as a useful tool for cytotechnological manipulation and transgenic modification of this fish.

  19. Potential of mean force calculation of the free energy of adsorption of Type I winter flounder antifreeze protein on ice

    NASA Astrophysics Data System (ADS)

    Battle, Keith; Alan Salter, E.; Wesley Edmunds, R.; Wierzbicki, Andrzej

    2010-04-01

    Antifreeze proteins (AFPs) are a unique class of proteins that inhibit ice growth without changing the melting point of ice. In this work, we study the detailed molecular mechanism of interactions between the hydrophobic side of the winter flounder (WF) AFP and two mutants, AAAA and SSSS, in which threonine residues are substituted by serines and alanines, respectively. Umbrella sampling molecular dynamics simulations of the separation of the proteins from the (2 0 1) surface in an explicit water box is carried out to calculate the potential of mean force free energies of adsorption using AMBER10i. We estimate wild-type WF's free energy of adsorption to ice to be about -12.0 kcal/mol. Gas-phase pseudopotential plane-wave calculations of methane adsorption onto select surfaces of ice are also carried out under periodic boundary conditions to address the possible enthalpic role of WF's methyl groups in binding. The contributions of hydrophobic residues to the free energy of adsorption are discussed.

  20. Molecular Cloning, Promoter Analysis and Expression Profiles of the sox3 Gene in Japanese Flounder, Paralichthys olivaceus

    PubMed Central

    Gao, Jinning; Li, Peizhen; Zhang, Wei; Wang, Zhigang; Wang, Xubo; Zhang, Quanqi

    2015-01-01

    Sox3, which belongs to the SoxB1 subgroup, plays major roles in neural and gonadal development. In the present study, Japanese flounder Paralichthys olivaceus sox3 gene (Posox3) and its promoter sequence were isolated and characterized. The deduced PoSox3 protein contained 298 amino acids with a characteristic HMG-box domain. Alignment and phylogenetic analyses indicated that PoSox3 shares highly identical sequence with Sox3 homologues from different species. The promoter region of Posox3 has many potential transcription factor (TF) binding sites. The expression profiles of Posox3 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR (qRT-PCR). Posox3 mRNA was maternally inherited, and maintained at a considerably high expression level between the blastula stage and the hatching stage during embryonic development. Posox3 was abundantly expressed in the adult brain and showed sexually dimorphic expression pattern. In situ hybridization (ISH) was carried out to investigate the cellular distribution of Posox3 in the ovary, and results showed the uniform distribution of Posox3 throughout the cytoplasm of oogonia and stage I–III oocytes. These results indicate that Posox3 has potentially vital roles in embryonic and neural development and may be involved in the oogenesis process. Our work provides a fundamental understanding of the structure and potential functions of Sox3 in Paralichthys olivaceus. PMID:26610486

  1. Fine-scale movements and habitat use of juvenile southern flounder Paralichthys lethostigma in an estuarine seascape.

    PubMed

    Furey, N B; Dance, M A; Rooker, J R

    2013-05-01

    Habitat use of juvenile southern flounder Paralichthys lethostigma was examined within a shallow estuarine seascape during June and July 2011 using acoustic telemetry. Fine-scale movement and habitat use of P. lethostigma was investigated with an acoustic positioning system placed in a seascape that varied in habitat type, physicochemical conditions and bathymetry. The use of different habitat types was examined with Euclidean distance-based analyses, and generalized additive models were used to determine the relative importance of habitat type relative to physicochemical conditions and bathymetry. Tracks of P. lethostigma ranged in distance between 1477 and 8582 m and speed was 4·2 ± 1·1 m min⁻¹ (mean ± s.e.) for all P. lethostigma combined. Depth, slope and habitat type had the most influence on P. lethostigma occurrence and deep sandy areas with shallow slopes were used most frequently. In addition, depth use by P. lethostigma was influenced by tidal cycles, indicating habitat use varies temporally and is dynamic. Finally, temperatures <30·5° C were used more than warmer waters within the study area. The results successfully identify movements by juvenile P. lethostigma, and indicate that definitions of essential habitats need to account for dynamics in habitat use.

  2. Do early growth dynamics explain recruitment success in Japanese flounder Paralichthys olivaceus off the Pacific coast of northern Japan?

    NASA Astrophysics Data System (ADS)

    Oshima, Masakane; Robert, Dominique; Kurita, Yutaka; Yoneda, Michio; Tominaga, Osamu; Tomiyama, Takeshi; Yamashita, Yoh; Uehara, Shinji

    2010-07-01

    We examined the relative importance of growth-related processes operating during the larval and early juvenile stage in explaining variability in year class success of Japanese flounder off the Pacific coast of northern Japan. Early growth trajectory of larvae and juveniles sampled in 2005 (strong year class) and in 2006 (weak year class) was estimated from the analysis of the lapillar otolith. The larval pelagic stage was characterized by lower growth and developmental rates, as well as high selection for fast growth in the metamorphosis/settlement period, during the strong recruitment event of 2005 relative to 2006. Growth appeared higher in 2005 only after settlement despite high density in the nursery, which likely reflected superior productivity during that year combined with an increased probability of cannibalism from early settlers on late settlers. This implies that larval growth dynamics did not play an important role in determining recruitment strength in the two years considered. The decreasing distance from the nursery areas of pelagic larvae through ontogeny in 2005, combined with low age at settlement, suggest that larvae benefited from positive transport conditions during the dominant year class. To the contrary, unfavorable hydrographic conditions likely prevailed in 2006 as distance from the nurseries increased with ontogeny and settlement occurred later than in 2005 despite faster growth potential and developmental rate. We conclude that transport conditions to the nursery grounds, rather than larval growth potential, represented the most important determinant of year class success in the two years considered.

  3. The duplicated paired box protein 7 (pax7) genes differentially transcribed during Japanese flounder (Paralichthys olivaceus) embryogenesis.

    PubMed

    Jiao, Shuang; Tan, Xungang; Li, Meijie; Sui, Yulei; Du, Shao Jun; You, Feng

    2015-11-01

    PAX are important regulators of developmental processes. PAX7 plays crucial roles in patterning of the dorsal central nervous system (CNS), neural crest (NC), and skeletal muscle. Here, we identified six spliced isoforms of pax7a and one pax7b and characterized their expression patterns. All of flounder Pax7a-1, Pax7a-2, Pax7a-3, and Pax7b contain a conserved paired domain (PD), an octapeptide motif (OP), and a paired type homeodomain (HD). However, the PD of Pax7a-4 and the HD of Pax7a-5 are not intact, and there is no HD in Pax7a-4 and Pax7a-6. pax7a and pax7b show distinct spatiotemporal expression patterns during embryogenesis. Whole-mount in situ hybridization demonstrates that the expression patterns of pax7a and pax7b are overlapping but distinguishable in the dorsal central nervous system. pax7a is expressed in most part of the brain and the neural tube, while pax7b is expressed exclusively in the diencephalon and the midbrain. In addition, pax7a is also expressed in the cranial NC and the trunk NC. RT-PCR results show that there were different expression patterns between the different isoforms. These results indicate subfunction partitioning of the duplicated pax7 genes. The duplicated pax7 may provide additional flexibility in fine-tuning neurogenesis and somitogenesis.

  4. Molecular Cloning, Promoter Analysis and Expression Profiles of the sox3 Gene in Japanese Flounder, Paralichthys olivaceus.

    PubMed

    Gao, Jinning; Li, Peizhen; Zhang, Wei; Wang, Zhigang; Wang, Xubo; Zhang, Quanqi

    2015-01-01

    Sox3, which belongs to the SoxB1 subgroup, plays major roles in neural and gonadal development. In the present study, Japanese flounder Paralichthys olivaceus sox3 gene (Posox3) and its promoter sequence were isolated and characterized. The deduced PoSox3 protein contained 298 amino acids with a characteristic HMG-box domain. Alignment and phylogenetic analyses indicated that PoSox3 shares highly identical sequence with Sox3 homologues from different species. The promoter region of Posox3 has many potential transcription factor (TF) binding sites. The expression profiles of Posox3 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR (qRT-PCR). Posox3 mRNA was maternally inherited, and maintained at a considerably high expression level between the blastula stage and the hatching stage during embryonic development. Posox3 was abundantly expressed in the adult brain and showed sexually dimorphic expression pattern. In situ hybridization (ISH) was carried out to investigate the cellular distribution of Posox3 in the ovary, and results showed the uniform distribution of Posox3 throughout the cytoplasm of oogonia and stage I-III oocytes. These results indicate that Posox3 has potentially vital roles in embryonic and neural development and may be involved in the oogenesis process. Our work provides a fundamental understanding of the structure and potential functions of Sox3 in Paralichthys olivaceus. PMID:26610486

  5. Development of the GENIPOL European flounder (Platichthys flesus) microarray and determination of temporal transcriptional responses to cadmium at low dose.

    PubMed

    Williams, Tim D; Diab, Amer M; George, Stephen G; Godfrey, Rita E; Sabine, Victoria; Conesa, Ana; Minchin, Steven D; Watts, Phil C; Chipman, James K

    2006-10-15

    We have constructed a high density, 13 270-clone cDNA array for the sentinel fish species European flounder (Platichthys flesus), combining clones from suppressive subtractive hybridization and a liver cDNA library; DNA sequences of 5211 clones were determined. Fish were treated by single intraperitoneal injection with 50 micrograms cadmium chloride per kilogram body weight, a dose relevant to environmental exposures, and hepatic gene expression changes were determined at 1, 2, 4, 8, and 16 days postinjection in comparison to saline-treated controls. Gene expression responses were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR). Blast2GO gene ontology analysis highlighted a general induction of the unfolded protein response, response to oxidative stress, protein synthesis, transport, and degradation pathways, while apoptosis, cell cycle, cytoskeleton, and cytokine genes were also affected. Transcript levels of cytochrome P450 1A (CYP1A) were repressed and vitellogenin altered, real-time PCR showed induction of metallothionein. We thus describe the establishment of a useful resource for ecotoxicogenomics and the determination of the temporal molecular responses to cadmium, a prototypical heavy metal pollutant.

  6. The effects of temperature on ovarian aromatase (cyp19a1a) expression and sex differentiation in summer flounder (Paralichthys dentatus).

    PubMed

    Caruso, Catherine C; Breton, Timothy S; Berlinsky, David L

    2016-04-01

    Female summer flounder grow considerably faster and larger than males, and a tremendous increase in performance can therefore be realized through production of monosex female populations. Rearing temperature has been shown to affect sex differentiation in other teleost species by influencing expression of genes encoding transcription factors or enzymes involved in endocrine function. Cyp19a1a is a well-studied gene that had been shown to play a role in ovarian development, and exhibits sexually dimorphic expression in other species. In the present study, summer flounder (37 days post-hatch; DPH) were raised at 13, 16 or 19 °C. Fish from all three treatments were sampled throughout development and analyzed in qPCR to determine cyp19a1a gene expression levels. Sex ratios of additional fish grown to ≥150 mm at each temperature treatment were determined. Low female production was achieved overall (26.9, 17.6 and 0% at 13, 16 and 19 °C, respectively). Cyp19a1a expression was significantly lower at 52 DPH (~15 mm total length) at the male-producing temperature (19 °C) and increased to similar levels as other treatments at 66 DPH. Expression levels later in juvenile development (66-191 DPH) largely decreased with fish size. The period of sex differentiation in summer flounder remains unknown, but cyp19a1a expression patterns suggest that it may occur earlier in development than that of congenerics. Further research is necessary to understand the sex-determining mechanisms in this species before sexually dimorphic growth can be used to achieve economic advantages in commercial production.

  7. Distinct immune tones are established by Lactococcus lactis BFE920 and Lactobacillus plantarum FGL0001 in the gut of olive flounder (Paralichthys olivaceus).

    PubMed

    Beck, Bo Ram; Song, Joon Hyun; Park, Byung Sun; Kim, Daniel; Kwak, Jin-Hwan; Do, Hyung Ki; Kim, Ah-Ram; Kim, Woo-Jin; Song, Seong Kyu

    2016-08-01

    The immune tone is defined as an immunological state during which the readiness for immune response is potentiated. The establishment of immune tone in the gut of olive flounder (Paralichthys olivaceus) was investigated by feeding Lactococcus lactis BFE920 (LL) or Lactobacillus plantarum FGL0001 (LP). LL-fed flounder showed significantly increased levels of regulatory genes (FOXP3, IL-10, and TGF-β1), CD18, and CD83 in the gut. In contrast, LP feeding drastically increased proinflammatory genes (T-bet, IL-1β, and IFN-γ) and CD18. This indicates that LL and LP establish different types of local immune tones in the gut through differential activation of innate immune cells: LL activates both macrophages and dendritic cells while LP activates macrophages only. Both of the immune tones required at least a total of 6 probiotic feeds during 72 h for a stable establishment. Once established, the type of immune tone remained steady even up to 30 days (a total of 60 feeds) probiotics feeding. The LL-induced regulatory immune tone enhanced the level of occludin, a tight junction molecule, significantly more than that observed with the proinflammatory immune tone established by LP feeding. Consequently, LL-fed fish showed considerably lower gut permeability than that of the LP-fed group. Furthermore, when orally challenged by Edwardsiella tarda, LL-fed flounder survived at a significantly higher rate than LP-fed fish. The data clearly demonstrate that individual probiotics establish distinct types of immune tone in the fish gut, which in turn influences the immunological status as well as the physiology of the gut. Selection of proper probiotics may be essential for optimal effects in aquaculture farming. PMID:27320869

  8. Expression of insulin-like growth factor I receptors at mRNA and protein levels during metamorphosis of Japanese flounder (Paralichthys olivaceus).

    PubMed

    Zhang, Junling; Shi, Zhiyi; Cheng, Qi; Chen, Xiaowu

    2011-08-01

    Insulin-like growth factor I (IGF-I) is an important regulator of fish growth and development, and its biological actions are initiated by binding to IGF-I receptor (IGF-IR). Our previous study has revealed that IGF-I could play an important role during metamorphosis of Japanese flounder, Paralichthys olivaceus. The analysis of IGF-IR expression thus helps further elucidate the IGF-I regulation of metamorphic processes. In this study, the spatial-temporal expression of two distinct IGF-IR mRNAs was investigated by real-time RT-PCR. The spatial distribution of two IGF-IR mRNAs in adult tissues is largely overlapped, but they exhibit distinct temporal expression patterns during larval development. A remarkable decrease in IGF-IR-2 mRNA was detected during metamorphosis. In contrast, a significant increase in IGF-IR-1 mRNA was determined from pre-metamorphosis to metamorphic completion. These indicate that they may play different function roles during the flounder metamorphosis. The levels and localization of IGF-IR proteins during larval development were further studied by Western blotting and immunohistochemistry. Immunoreactive IGF-IRs were detected throughout larval development, and the IGF-IR proteins displayed a relatively abundant expression during metamorphosis. Moreover, the IGF-IR proteins appeared in key tissues, such as thickened skin beneath the migrating eye, developing intestine, gills and kidney during metamorphosis. These results further suggest that the IGF-I system may be involved in metamorphic development of Japanese flounder.

  9. Disease prevalence in flounder (Platichthys flesus) from the Dutch Wadden Sea as indicator of environmental quality: A summary of 1988-2005 surveys

    NASA Astrophysics Data System (ADS)

    Vethaak, A. D.

    2013-09-01

    In 1988, epizootics of ulcer disease in the flatfish flounder in the Dutch Wadden Sea were reported near freshwater drainage sluices of IJsselmeer Lake, locally affecting up to 38.9% of fish. Other diseases such as fin rot and lymphocystis were less frequent, but followed a similar pattern. Results of follow-up surveys in the Wadden Sea in 1994-2005 confirm previous findings and also show significantly elevated ulcer prevalences at other smaller drainage works. The most likely stress factors that contributed to the development of the epizootics at these sites include osmotic stress, adverse water quality conditions including chemical contaminants, nutritional deficiencies, and obstruction to fish migration. It was shown that discharges of IJsselmeer Lake freshwater in 1988-96 had a wide effect on the prevalence and distribution of ulcers and lymphocystis in the western Wadden Sea. A general reduction in disease prevalence in flounder in the entire Dutch Wadden Sea was observed during 1988-2005, which was most likely due to a general improvement in water quality and locally improved habitat conditions for flounder near drainage sluices. Ulcer prevalences outside the two IJsselmeer Lake sluices (Den Oever and Kornwerderzand) declined in this period from approximately 30% to 10% for medium-sized fish. Other skin diseases have also displayed a downward trend at both sites in recent years, with prevalences falling sharply to below 1%. Elsewhere in the Wadden Sea and the Ems-Dollard estuary, disease prevalences have declined towards natural background levels (< 1%). It is concluded that skin diseases, especially ulcers, are useful indicators of environmental quality in the Wadden Sea.

  10. Cloning of the cDNA encoding adenosine 5'-monophosphate deaminase 1 and its mRNA expression in Japanese flounder Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Jiang, Keyong; Sun, Shujuan; Liu, Mei; Wang, Baojie; Meng, Xiaolin; Wang, Lei

    2013-01-01

    AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5'-UTR of 78 bp, a 3'-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) ( P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) ( P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.

  11. Immigration and early life stages recruitment of the European flounder (Platichthys flesus) to an estuarine nursery: The influence of environmental factors

    NASA Astrophysics Data System (ADS)

    Amorim, Eva; Ramos, Sandra; Elliott, Michael; Bordalo, Adriano A.

    2016-01-01

    Connectivity between coastal spawning grounds and estuarine nurseries is a critical step in the life cycle of many fish species. Larval immigration and transport-associated physical-biological processes are determinants of recruitment success to nursery areas. The recruitment of the European flounder, Platichthys flesus, to estuarine nurseries located at the southern edge of the species distribution range, has been usually investigated during its juvenile stages, while estuarine recruitment during the earlier planktonic life stage remains largely unstudied. The present study investigated the patterns of flounder larval recruitment and the influence of environmental factors on the immigration of the early life stages to the Lima estuary (NW Portugal), integrating data on fish larvae and post-settlement individuals (< 50 mm length), collected over 7 years. Late-stage larvae arrived at the estuary between February and July and peak abundances were observed in April. Post-settlement individuals (< 50 mm) occurred later between April and October, whereas newly-settled ones (< 20 mm) were found only in May and June. Variables associated with the spawning, survival and growth of larvae in the ocean (sea surface temperature, chlorophyll a and inland hydrological variables) were the major drivers of flounder occurrence in the estuarine nursery. Although the adjacent coastal area is characterized by a current system with strong seasonality and mesoscale variability, we did not identify any influence of variables related with physical processes (currents and upwelling) on the occurrence of early life stages in the estuary. A wider knowledge on the influence of the coastal circulation variability and its associated effects upon ocean-estuarine connectivity is required to improve our understanding of the population dynamics of marine spawning fish that use estuarine nurseries.

  12. Survey of estrogenic activity in United Kingdom estuarine and coastal waters and its effects on gonadal development of the flounder Platichthys flesus

    SciTech Connect

    Allen, Y.; Scott, A.P.; Matthiessen, P.; Haworth, S.; Thain, J.E.; Feist, S.

    1999-08-01

    Estrogenic discharges are known or suspected to be present in many United Kingdom estuaries and are also occurring from offshore oil and gas installations. The aim of this study was to establish whether estrogens and their mimics are present in marine waters at concentrations that can produce biological responses in fish and, if so, to discover whether the effects are likely to be harmful to populations and communities through changes in reproductive and other physiological processes. Laboratory studies and extensive field surveys were carried out with the euryhaline flounder (Platichthys flesus), using the amount of yolk protein vitellogenin in the blood of free-living male fish as an indicator of exposure to estrogens. Exposure to the synthetic estrogen ethynylestradiol for 3 weeks confirmed that the vitellogenin response was made and showed that this species is about one order of magnitude less sensitive than the freshwater species rainbow trout. The fish did not respond to the estrogen mimic nonylphenol at concentrations of 30 {micro}g/L. Wild male flounder were sampled from four contaminated United Kingdom estuaries: the Tyne, Crouch, Thames, and Mersey. Fish from one or more sites in each estuary had significantly elevated concentrations of vitellogenin in their blood when compared to concentrations in fish from the clean control estuary (Alde River). Vitellogenin concentrations in the Tyne and Mersey fish were four and six orders of magnitude greater, respectively, than control concentrations. Elevated vitellogenin concentrations were also seen in some male flounder caught in coastal areas. Approximately 20% of male fish from the Mersey estuary contained oocytes in their testes, but this intersex condition was not seen elsewhere. The implications of these findings for fish populations are discussed.

  13. Surveys of plasma vitellogenin and intersex in male flounder (Platichthys flesus) as measures of endocrine disruption by estrogenic contamination in United Kingdom estuaries: temporal trends, 1996 to 2001.

    PubMed

    Kirby, Mark F; Allen, Yvonne T; Dyer, Robert A; Feist, Steve W; Katsiadaki, Ioanna; Matthiessen, Peter; Scott, Alex P; Smith, Andrew; Stentiford, Grant D; Thain, John E; Thomas, Kevin V; Tolhurst, Laura; Waldock, Michael J

    2004-03-01

    Plasma vitellogenin (VTG) concentrations and the presence of the ovo-testis (intersex) condition have been recorded in male flounder (Platichthys flesus) captured from several United Kingdom (UK) estuaries since 1996 as part of the endocrine disruption in the Marine Environment (EDMAR) project and earlier programs. It has been confirmed that plasma VTG concentrations in male flounder have remained elevated in several UK estuaries (e.g., Tees, Mersey, and Tyne) throughout the period covered by this study. However, the time-series data indicate that plasma VTG, a measure of environmental estrogen contamination, has decreased in fish captured from several estuaries, especially those of the Tyne and Mersey. Shorter time-series data sets from the Forth and Clyde estuaries also suggest a decrease in estrogen contamination at these sites. Trends associated with specific point sources of estrogenic contamination show site-specific patterns. For instance, plasma VTG levels in male flounder captured near the Howdon sewage treatment outfall (Tyne) have shown a steady decline to near baseline levels in 2001, while the plasma of male fish captured at a site adjacent to the Dabholm Gut discharge in the Tees estuary have shown little evidence of a sustained decline. The occurrence of the intersex condition was detected at a low but consistent prevalence through the study period, with the majority of cases recorded in fish captured from the Tyne and Mersey estuaries. The data set does not allow conclusions to be drawn about any temporal trends associated with this condition. The significance of the findings and possible mitigating influences are discussed in terms of the impacts on wild fish and the role of effluent treatment in reducing these.

  14. Toxicity of analytically cleaned pentabromodiphenylether after prolonged exposure in estuarine European flounder (Platichthys flesus), and partial life-cycle exposure in fresh water zebrafish (Danio rerio).

    PubMed

    Kuiper, Raoul V; Vethaak, A D; Cantón, Roćio F; Anselmo, Henrique; Dubbeldam, Marco; van den Brandhof, Evert-Jan; Leonards, Pim E G; Wester, Piet W; van den Berg, Martin

    2008-09-01

    Residues of polybrominated diphenylethers (PBDEs), extensively applied as flame retardants, are widely spread in the aquatic environment and biota. The present study investigates effects of the environmentally relevant lower brominated diphenylethers in two fish species in vivo under controlled laboratory conditions. Euryhaline flounder (Platichthys flesus) and freshwater zebrafish (Danio rerio) were exposed to a range of concentrations of a commercial pentabromodiphenylether mixture, DE-71. Chemical analysis of exposed fish showed a pattern of PBDE congeners that was very similar to that in wild fish. The resulting range included environmentally relevant, as well as higher levels. Animals were investigated histopathologically with emphasis on endocrine and reproductive organs. In zebrafish, hatching of embryos and larval development were assessed. Biochemical parameters were investigated in flounder as markers for suggested dioxin-like activity (ethoxyresorufin-O-deethylase=EROD), and activation of endogenous estrogen synthesis (gonad aromatase activity). Thyroid hormones were analyzed in plasma in both species. Benchmark analysis using internal PBDE concentrations showed a mild dose-dependent decrease of hepatic EROD and ovarian aromatase activities, and plasma thyroxin levels in flounder, and an increase of plasma thyroid hormone levels in zebrafish. These trends did not result in statistically significant differences from control fish, and major histopathological changes were not observed. Reproduction in zebrafish appeared to be the most sensitive parameter with statistically significantly reduced larval survival and non-significant indications for decreased egg production at internal levels that were more than 55 times the highest environmental recordings. The present results indicate limited risk for endocrine or reproductive effects of current environmental PBDE contamination in fish.

  15. Distinct immune tones are established by Lactococcus lactis BFE920 and Lactobacillus plantarum FGL0001 in the gut of olive flounder (Paralichthys olivaceus).

    PubMed

    Beck, Bo Ram; Song, Joon Hyun; Park, Byung Sun; Kim, Daniel; Kwak, Jin-Hwan; Do, Hyung Ki; Kim, Ah-Ram; Kim, Woo-Jin; Song, Seong Kyu

    2016-08-01

    The immune tone is defined as an immunological state during which the readiness for immune response is potentiated. The establishment of immune tone in the gut of olive flounder (Paralichthys olivaceus) was investigated by feeding Lactococcus lactis BFE920 (LL) or Lactobacillus plantarum FGL0001 (LP). LL-fed flounder showed significantly increased levels of regulatory genes (FOXP3, IL-10, and TGF-β1), CD18, and CD83 in the gut. In contrast, LP feeding drastically increased proinflammatory genes (T-bet, IL-1β, and IFN-γ) and CD18. This indicates that LL and LP establish different types of local immune tones in the gut through differential activation of innate immune cells: LL activates both macrophages and dendritic cells while LP activates macrophages only. Both of the immune tones required at least a total of 6 probiotic feeds during 72 h for a stable establishment. Once established, the type of immune tone remained steady even up to 30 days (a total of 60 feeds) probiotics feeding. The LL-induced regulatory immune tone enhanced the level of occludin, a tight junction molecule, significantly more than that observed with the proinflammatory immune tone established by LP feeding. Consequently, LL-fed fish showed considerably lower gut permeability than that of the LP-fed group. Furthermore, when orally challenged by Edwardsiella tarda, LL-fed flounder survived at a significantly higher rate than LP-fed fish. The data clearly demonstrate that individual probiotics establish distinct types of immune tone in the fish gut, which in turn influences the immunological status as well as the physiology of the gut. Selection of proper probiotics may be essential for optimal effects in aquaculture farming.

  16. Effects of calanoid copepod Schmackeria poplesia as a live food on the growth, survival and fatty acid composition of larvae and juveniles of Japanese flounder, Paralichthys olivaceus

    NASA Astrophysics Data System (ADS)

    Liu, Guangxing; Xu, Donghui

    2009-12-01

    Zooplankton constitutes a major part of the diet for fish larvae in the marine food web, and it is generally believed that copepods can meet the nutritional requirements of fish larvae. In this study, calanoid copepod Schmackeria poplesia, rotifer Brachionus plicatilis and anostraca crustacean Artemia sp. were analyzed for fatty acid contents, and were used as live food for culturing larval Japanese flounder, Paralichthys olivaceus. The total content of three types of HUFAs (DHA, EPA and ARA) in S. poplesia was significantly higher than that in the other two live foods ( P<0.01). Three live organisms were used for raising larvae and juveniles of Paralichthys olivaceus respectively for 15 and 10 d. Then the growth, survival and fatty acid composition of the larvae and juveniles were investigated. The results showed that the larvae and juveniles fed with copepods ( S. poplesia) had significantly higher growth rate than those fed with the other two organisms ( P<0.01). The survival of the flounder larvae fed with copepods was significantly higher than that of the others ( P<0.01), and the survival of the juvenile fish fed with copepods was higher than that fed with Artemia ( P<0.05). The contents of three types of HUFAs (DHA, EPA and ARA) and the ratio of DHA/EPA in larval and juvenile flounder P. olivaceus were analyzed. The results showed that the contents of DHA, EPA and ARA in the larvae and juveniles fed with S. poplesia were higher than those fed with a mixed diet or Artemia only, and the ratio of EPA/ARA in larvae and juveniles of P. olivaceus fed with S. poplesia was lower than that in the case of feeding with a mixed diet or Artemia only. The present data showed that copepod is the best choice for feeding the larvae and juveniles of fish considering its effects on the survival, growth and nutrition composition of the fish.

  17. Identification of a Tc1-like transposon integration site in the genome of the flounder (Platichthys flesus): a novel use of an inverse PCR method.

    PubMed

    Poćwierz-Kotus, Anita; Burzyński, Artur; Wenne, Roman

    2010-03-01

    The inverse PCR method has been developed and applied employed for the identification of the integration sites of the Tc1-like transposons in the genome of the flounder, Platichthys flesus. One Tc1-like insertion instance was recognized and characterized, demonstrating an efficiency of the method for determining of transposon integration sites. The similarity of the sequence flanking transposon (SFT) to reverse transcriptase sequences (RVT) was demonstrated. It is likely that the insertion took place within currently degenerated LINE (long interspersed nuclear elements) retrotransposon.

  18. Molecular Characterization and Expression Analysis of ATP-Gated P2X7 Receptor Involved in Japanese Flounder (Paralichthys olivaceus) Innate Immune Response

    PubMed Central

    Li, Shuo; Li, Xuejing; Coddou, Claudio; Geng, Xuyun; Wei, Junli; Sun, Jinsheng

    2014-01-01

    ATP-gated P2X7 receptor (P2RX7) channel is a key component for purinergic signaling and plays important roles in the innate immune response in mammals. However, the expression, molecular properties and immune significances of P2RX7 in lower vertebrates are still very limited. Here we identified and characterized a novel bony fish P2RX7 homologue cDNA, termed poP2RX7, in Japanese flounder (Paralichthys olivaceus). PoP2RX7 protein shares about 60–88% sequence similarity and 45–78% sequence identity with known vertebrate P2RX7 proteins. Phylogenetic analysis placed poP2RX7 and other P2RX7 proteins within their own cluster apart from other P2RX members. While the functional poP2RX7 channel shares structural features in common with known P2RX7 homologs, electrophysiological studies revealed that BzATP, the more potent agonist for known mammalian and fish P2RX7s, shows similar potency to ATP in poP2RX7 activation. poP2RX7 mRNA constitutively expressed in all examined tissues from unstimulated healthy Japanese flounder with dominant expression in hepatopancreas and the lowest expression in head kidney, trunk kidney, spleen and gill. poP2RX7 mRNA expression, however, was significantly induced in Japanese flounder head kidney primary cells by Poly(I:C) and bacterial endotoxin LPS stimulations. In vivo experiments further revealed that poP2RX7 gene expression was substantially up-regulated by immune challenge with infectious bacteria Edwardsiella tarda and Vibrio anguillarum. Moreover, activation of poP2RX7 results in an increased gene expression of multifunctional cytokines IL-1β and IL-6 in the head kidney primary cells. Collectively, we identified and characterized a novel fish P2RX7 homolog which is engaged in Japanese flounder innate immune response probably through modulation of pro-inflammatory cytokines expression. PMID:24796752

  19. Skin and liver diseases induced in flounder (Platichthys flesus) after long-term exposure to contaminated sediments in large-scale mesocosms.

    PubMed Central

    Vethaak, A D; Jol, J G; Meijboom, A; Eggens, M L; Rheinallt, T; Wester, P W; van de Zande, T; Bergman, A; Dankers, N; Ariese, F; Baan, R A; Everts, J M; Opperhuizen, A; Marquenie, J M

    1996-01-01

    Disease development in flounder (Platichthys flesus) was studied over a period of 3 years in three large mesocosms (40 m x 40 m x 3 m). Two of the mesocosms contained clean sand and the third, sharing a common water circulation with one of the clean-sand mesocosms, was stocked with contaminated dredged spoil. In this way, one of the clean-sand mesocosms was indirectly polluted via the water phase, and analysis of contaminant concentrations in sediments and flounder tissues showed that it had a status intermediate between the other two. Random samples of the flounder populations from the indirectly polluted and reference mesocosms were examined every 2 months for epidermal diseases (lymphocystis, skin ulcers, fin rot) and then released. In addition, every 6 months, random samples of fish from all three mesocosms were sacrificed for histological and chemical investigation. With regard to the development of epidermal disease, the results showed little difference between the reference mesocosm and the indirectly polluted mesocosm, with the exception that lymphocystis was significantly elevated in the indirectly polluted mesocosm. Although pollution may be a risk factor in the etiology of this disease, such a relationship would probably be obscured under field conditions due to variation arising from other factors. Histopathological analysis of the livers revealed in total four cases of hepatocellular adenoma (1.5% of sampled population) in fish from the polluted mesocosms, the first occurring after 2.5 years of exposure in fish from the indirectly polluted mesocosm. Furthermore, several other liver lesions, including foci of cellular alteration and hydropic vacuolated lesions, developed during the course of the experiment before tumor formation was apparent. Prevalences of these conditions were very much lower in the reference mesocosm than in the two polluted mesocosms. Densities of melanomacrophage centers in the liver showed a similar trend. The findings clearly

  20. Identification of arsenobetaine in sole, lemon sole, flounder, dab, crab and shrimps by field desorption and fast atom bombardment mass spectrometry

    SciTech Connect

    Luten, J.B.; Riekwel-Booy, G.; Greef, M.C.; de Brauw, I.N.

    1983-01-01

    Organo-arsenic has been isolated from sole, lemon sole, flounder, dab, crab and shrimps by extraction or ion-exchange in combination with thin-layer chromatography. An alkaline digestion of the samples, followed by a reduction with sodiumborohydride leads to the formation of trimethylarsine. Field desorption mass spectrometry (FDMS) can be used to identify arsenobetaine in the isolates. Sufficient purification by thin-layer chromatography is found to be a prerequisite for the detection of a protonated molecular ion of arsenobetaine. If this situation is not met acid enchanced FDMS or Fast Atom Bombardment mass spectrometry in high resolution can be used successfully.

  1. Factors Affecting Mercury and Selenium Levels-in New Jersey Flatfish: Low Risk to Human Consumers

    PubMed Central

    Burger, Joanna; Jeitner, Christian; Donio, Mark; Shukla, Sheila; Gochfeld, Michael

    2014-01-01

    Some fish contain high levels of mercury (Hg), which could pose a risk to fish eaters themselves or their children. In making decisions about fish consumption, people must decide whether to eat fish, how much to eat, what species to eat, and what size fish to eat, as well as suitable (or unsuitable) locations, among other factors. Yet to make sound decisions, people need to know the levels of Hg in fish as a function of species, size, and location of capture. Levels of Hg and selenium (Se) were examined in three species of flatfish (fluke or summer flounder [Paralichthys dentatus], winter flounder [Pseudopleuronectes americanus], and windowpane [Scophthalmus aquosus]) from New Jersey as a function of species, fish size, season, and location. Flatfish were postulated to have low levels of Hg because they are low on the food chain and are bottom feeders, and data were generated to provide individuals with information on a species that might be safe to eat regularly. Although there were interspecific differences in Hg levels in the 3 species, total Hg levels averaged 0.18, 0.14, and 0.06 ppm (μg/g, wet weigh) in windowpane, fluke, and winter flounder, and selenium levels averaged 0.36, 0.35, and 0.25 ppm, respectively. For windowpane, 15% had Hg levels above 0.3 ppm, but no individual fish had Hg levels over 0.5 ppm. There were no significant seasonal differences in Hg levels, although Se was significantly higher in fluke in summer compared to spring. There were few geographical differences among New Jersey locations. Correlations between Hg and Se levels were low. Data, based on 464 fish samples, indicate that Hg levels are below various advisory levels and pose little risk to typical New Jersey fish consumers. A 70-kg person eating 1 meal (8 oz or 227 g) per week would not exceed the U.S. Environmental Protection Agency reference dose of 0.1 μg/kg body weight/d of methylmercury (MeHg). However, high-end fish eaters consuming several such meals per week may exceed

  2. Isolation and genetic characterization of Toxoplasma gondii from raccoons (Procyon lotor), cats (Felis domesticus), striped skunk (Mephitis mephitis), black bear (Ursus americanus), and cougar (Puma concolor) from Canada.

    PubMed

    Dubey, J P; Quirk, T; Pittt, J A; Sundar, N; Velmurugan, G V; Kwok, O C H; Leclair, D; Hill, R; Su, C

    2008-02-01

    Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats (Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus) from Kuujjuaq, northern Quebec, Canada. Genotyping of these T. gondii isolates using polymorphisms at 10 nuclear markers including SAGI, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico revealed 4 genotypes. None of the isolates was clonal archetypal Types I, II, and III found in the United States. These results are in contrast with the Type II genotype that is widespread in domestic animals and humans throughout the United States and Europe. This is the first genotyping of T. gondii isolates from this part of North America.

  3. Triplex polymerase chain reaction assay for detection of major soil-transmitted helminths, Ascaris lumbricoides, Trichuris trichiura, Necator americanus, in fecal samples.

    PubMed

    Phuphisut, Orawan; Yoonuan, Tippayarat; Sanguankiat, Surapol; Chaisiri, Kittipong; Maipanich, Wanna; Pubampen, Somchit; Komalamisra, Chalit; Adisakwattana, Poom

    2014-03-01

    Ascaris lumbricoides, Trichuris trichiura, and Necator americanus are medically important soil-transmitted helminths (STHs) occurring frequently worldwide including Thailand. Fecal examination using a microscope has been recommended as the gold standard for diagnosis of STH infections, but suffers from low sensitivity. Recently, highly sensitive and specific assays, such as multiplex quantitative PCR, has been established, but the high cost and need for special instruments are still barriers limiting their applications in routine diagnosis. Therefore, a conventional multiplex PCR assay, with its lower cost and greater simplicity, was developed, for the simultaneous detection of STHs in fecal samples. The multiplex PCR assay was species-specific to the three STHs, and could detect one copy of DNA target. Compared with microscopic examination of fecal samples, sensitivity and specificity of the multiplex PCR was 87% and 83%, respectively. This multiplex PCR assay provides an alternative method for routine diagnosis of STHs infection, and might be applied for epidemiological studies of STHs in endemic areas.

  4. Structural and mechanical properties of the arthropod cuticle: comparison between the fang of the spider Cupiennius salei and the carapace of American lobster Homarus americanus.

    PubMed

    Erko, Maxim; Hartmann, Markus A; Zlotnikov, Igor; Valverde Serrano, Clara; Fratzl, Peter; Politi, Yael

    2013-08-01

    Most biological materials are nanocomposites characterized by a multi-level structural hierarchy. Particularly, the arthropod cuticle is a chitin-based composite material where the mechanical properties strongly depend on both molecular chitin/protein properties, and the structural arrangement of chitin-fibrils within the protein matrix. Here materials properties and structural organization of two types of cuticle from distantly related arthropods, the wandering spider Cupiennius salei and American lobster Homarus americanus were studied using nanoindentation and X-ray diffraction. The structural analysis of the two types of cuticle including the packing and alignment of chitin-fibrils is supported by Monte Carlo simulations of the experimental X-ray data, thereby regions of parallel and rotated fibril arrangement can be clearly distinguished. The tip of the spider fang which is used to inject venom into the prey was found to be considerably harder than the lobster carapace, while its stiffness is slightly lower. PMID:23765087

  5. Sequence conservation in the Ancylostoma secreted protein-2 of Necator americanus (Na-ASP-2) from hookworm infected individuals in Thailand.

    PubMed

    Ungcharoensuk, Charoenchai; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2012-12-01

    The Ancylostoma secreted protein-2 of Necator americanus (Na-ASP-2) was one of the promising vaccine candidates against the most prevalent human hookworm species as adverse vaccine reaction has compromised further human vaccine trials. To elucidate the gene structure and the extent of sequence diversity, we determined the complete nucleotide sequence of the Na-asp-2 gene of individual larvae from 32 infected subjects living in 3 different endemic areas of Thailand. Sequence analysis revealed that the gene encoding Na-ASP-2 comprised 8 exons. Of 3 nucleotide substitutions in these exons, only one causes an amino acid change from leucine to methionine. A consensus conserved GT and AG at the 5' and the 3' boundaries of each intron was observed akin to those found in other eukaryotic genes. Introns of Na-asp-2 contained 23 nucleotide substitutions and 0-18 indels. The mean number of nucleotide substitutions per site (d) in introns was not significantly different from the mean number of synonymous substitutions per synonymous site (d(S)) in exons whereas d in introns was significantly exceeded d(N) (the mean number of nonsynonymous substitutions per nonsynonymous site) in exons (p<0.05), suggesting that introns and synonymous sites in exons may evolve at a similar rate whereas functional constraints at the amino acid could limit amino acid substitutions in Na-ASP-2. A recombination site was identified in an intron near the 3' portion of the gene. The positions of introns and the intron phases in the Na-asp-2 gene comparing with those in other pathogenesis-related-1 proteins of Loa loa, Onchocerca volvulus, Heterodera glycines, Caenorhabditis elegans and human were relatively conserved, suggesting evolutionary conservation of these genes. Sequence conservation in Na-ASP-2 may not compromise further vaccine design if adverse vaccine effects could be resolved whereas microheterogeneity in introns of this locus may be useful for population genetics analysis of N. americanus

  6. Diversity in the structures and ligand-binding sites of nematode fatty acid and retinol-binding proteins revealed by Na-FAR-1 from Necator americanus

    PubMed Central

    Rey-Burusco, M. Florencia; Ibáñez-Shimabukuro, Marina; Gabrielsen, Mads; Franchini, Gisela R.; Roe, Andrew J.; Griffiths, Kate; Zhan, Bin; Cooper, Alan; Kennedy, Malcolm W.; Córsico, Betina; Smith, Brian O.

    2015-01-01

    Fatty acid and retinol-binding proteins (FARs) comprise a family of unusual α-helix rich lipid-binding proteins found exclusively in nematodes. They are secreted into host tissues by parasites of plants, animals and humans. The structure of a FAR protein from the free-living nematode Caenorhabditis elegans is available, but this protein [C. elegans FAR-7 (Ce-FAR-7)] is from a subfamily of FARs that does not appear to be important at the host/parasite interface. We have therefore examined [Necator americanus FAR-1 (Na-FAR-1)] from the blood-feeding intestinal parasite of humans, N. americanus. The 3D structure of Na-FAR-1 in its ligand-free and ligand-bound forms, determined by NMR (nuclear magnetic resonance) spectroscopy and X-ray crystallography respectively, reveals an α-helical fold similar to Ce-FAR-7, but Na-FAR-1 possesses a larger and more complex internal ligand-binding cavity and an additional C-terminal α-helix. Titration of apo-Na-FAR-1 with oleic acid, analysed by NMR chemical shift perturbation, reveals that at least four distinct protein–ligand complexes can be formed. Na-FAR-1 and possibly other FARs may have a wider repertoire for hydrophobic ligand binding, as confirmed in the present study by our finding that a range of neutral and polar lipids co-purify with the bacterially expressed recombinant protein. Finally, we show by immunohistochemistry that Na-FAR-1 is present in adult worms with a tissue distribution indicative of possible roles in nutrient acquisition by the parasite and in reproduction in the male. PMID:26318523

  7. Crystallization and preliminary X-ray analysis of Na-ASP-1, a multi-domain pathogenesis-related-1 protein from the human hookworm parasite Necator americanus

    SciTech Connect

    Asojo, Oluwatoyin A.; Loukas, Alex; Inan, Mehmet; Barent, Rick; Huang, Jicai; Plantz, Brad; Swanson, Amber; Gouthro, Mark; Meagher, Michael M.; Hotez, Peter J.

    2005-04-01

    In order to clarify the structural basis of the pathogenesis-related-1 domain, Na-ASP-1, the first multi-domain ASP from the human hookworm parasite N. americanus, has been crystallized. 2.2 Å resolution data have been collected from a crystal belonging to the monoclinic space group P2{sub 1}. Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis-related-1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been expressed, purified and crystallized. Na-ASP-1 is a 406-amino-acid multi-domain ASP from the prevalent human hookworm parasite Necator americanus. Useful X-ray data to 2.2 Å have been collected from a crystal that belongs to the monoclinic space group P2{sub 1} with unit-cell parameters a = 67.7, b = 74.27, c = 84.60 Å, β = 112.12°. An initial molecular-replacement solution has been obtained with one monomer in the asymmetric unit.

  8. An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection.

    PubMed

    Pearson, Mark S; Bethony, Jeffrey M; Pickering, Darren A; de Oliveira, Luciana M; Jariwala, Amar; Santiago, Helton; Miles, Aaron P; Zhan, Bin; Jiang, Desheng; Ranjit, Najju; Mulvenna, Jason; Tribolet, Leon; Plieskatt, Jordan; Smith, Tracey; Bottazzi, Maria Elena; Jones, Kathryn; Keegan, Brian; Hotez, Peter J; Loukas, Alex

    2009-09-01

    Hookworms digest hemoglobin from erythrocytes via a proteolytic cascade that begins with the aspartic protease, APR-1. Ac-APR-1 from the dog hookworm, Ancylostoma caninum, protects dogs against hookworm infection via antibodies that neutralize enzymatic activity and interrupt blood-feeding. Toward developing a human hookworm vaccine, we expressed both wild-type (Na-APR-1(wt)) and mutant (Na-APR-1(mut)-mutagenesis of the catalytic aspartic acids) forms of Na-APR-1 from the human hookworm, Necator americanus. Refolded Na-APR-1(wt) was catalytically active, and Na-APR-1(mut) was catalytically inactive but still bound substrates. Vaccination of canines with Na-APR-1(mut) and heterologous challenge with A. caninum resulted in significantly reduced parasite egg burdens (P=0.034) and weight loss (P=0.022). Vaccinated dogs also had less gut pathology, fewer adult worms, and reduced blood loss compared to controls but these did not reach statistical significance. Vaccination with Na-APR-1(mut) induced antibodies that bound the native enzyme in the parasite gut and neutralized enzymatic activity of Na-APR-1(wt) and APR-1 orthologues from three other hookworm species that infect humans. IgG1 against Na-APR-1(mut) was the most prominently detected antibody in sera from people resident in high-transmission areas for N. americanus, indicating that natural boosting may occur in exposed humans. Na-APR-1(mut) is now a lead antigen for the development of an antihematophagy vaccine for human hookworm disease.

  9. Sequence conservation in the Ancylostoma secreted protein-2 of Necator americanus (Na-ASP-2) from hookworm infected individuals in Thailand.

    PubMed

    Ungcharoensuk, Charoenchai; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2012-12-01

    The Ancylostoma secreted protein-2 of Necator americanus (Na-ASP-2) was one of the promising vaccine candidates against the most prevalent human hookworm species as adverse vaccine reaction has compromised further human vaccine trials. To elucidate the gene structure and the extent of sequence diversity, we determined the complete nucleotide sequence of the Na-asp-2 gene of individual larvae from 32 infected subjects living in 3 different endemic areas of Thailand. Sequence analysis revealed that the gene encoding Na-ASP-2 comprised 8 exons. Of 3 nucleotide substitutions in these exons, only one causes an amino acid change from leucine to methionine. A consensus conserved GT and AG at the 5' and the 3' boundaries of each intron was observed akin to those found in other eukaryotic genes. Introns of Na-asp-2 contained 23 nucleotide substitutions and 0-18 indels. The mean number of nucleotide substitutions per site (d) in introns was not significantly different from the mean number of synonymous substitutions per synonymous site (d(S)) in exons whereas d in introns was significantly exceeded d(N) (the mean number of nonsynonymous substitutions per nonsynonymous site) in exons (p<0.05), suggesting that introns and synonymous sites in exons may evolve at a similar rate whereas functional constraints at the amino acid could limit amino acid substitutions in Na-ASP-2. A recombination site was identified in an intron near the 3' portion of the gene. The positions of introns and the intron phases in the Na-asp-2 gene comparing with those in other pathogenesis-related-1 proteins of Loa loa, Onchocerca volvulus, Heterodera glycines, Caenorhabditis elegans and human were relatively conserved, suggesting evolutionary conservation of these genes. Sequence conservation in Na-ASP-2 may not compromise further vaccine design if adverse vaccine effects could be resolved whereas microheterogeneity in introns of this locus may be useful for population genetics analysis of N. americanus.

  10. Identification of a calcitonin-like diuretic hormone that functions as an intrinsic modulator of the American lobster, Homarus americanus, cardiac neuromuscular system.

    PubMed

    Christie, A E; Stevens, J S; Bowers, M R; Chapline, M C; Jensen, D A; Schegg, K M; Goldwaser, J; Kwiatkowski, M A; Pleasant, T K; Shoenfeld, L; Tempest, L K; Williams, C R; Wiwatpanit, T; Smith, C M; Beale, K M; Towle, D W; Schooley, D A; Dickinson, P S

    2010-01-01

    In insects, a family of peptides with sequence homology to the vertebrate calcitonins has been implicated in the control of diuresis, a process that includes mixing of the hemolymph. Here, we show that a member of the insect calcitonin-like diuretic hormone (CLDH) family is present in the American lobster, Homarus americanus, serving, at least in part, as a powerful modulator of cardiac output. Specifically, during an ongoing EST project, a transcript encoding a putative H. americanus CLDH precursor was identified; a full-length cDNA was subsequently cloned. In silico analyses of the deduced prepro-hormone predicted the mature structure of the encoded CLDH to be GLDLGLGRGFSGSQAAKHLMGLAAANFAGGPamide (Homam-CLDH), which is identical to a known Tribolium castaneum peptide. RT-PCR tissue profiling suggests that Homam-CLDH is broadly distributed within the lobster nervous system, including the cardiac ganglion (CG), which controls the movement of the neurogenic heart. RT-PCR analysis conducted on pacemaker neuron- and motor neuron-specific cDNAs suggests that the motor neurons are the source of the CLDH message in the CG. Perfusion of Homam-CLDH through the isolated lobster heart produced dose-dependent increases in both contraction frequency and amplitude and a dose-dependent decrease in contraction duration, with threshold concentrations for all parameters in the range 10(-11) to 10(-10) mol l(-1) or less, among the lowest for any peptide on this system. This report is the first documentation of a decapod CLDH, the first demonstration of CLDH bioactivity outside the Insecta, and the first detection of an intrinsic neuropeptide transcript in the crustacean CG. PMID:20008368

  11. Identification of a calcitonin-like diuretic hormone that functions as an intrinsic modulator of the American lobster, Homarus americanus, cardiac neuromuscular system

    PubMed Central

    Christie, A. E.; Stevens, J. S.; Bowers, M. R.; Chapline, M. C.; Jensen, D. A.; Schegg, K. M.; Goldwaser, J.; Kwiatkowski, M. A.; Pleasant, T. K.; Shoenfeld, L.; Tempest, L. K.; Williams, C. R.; Wiwatpanit, T.; Smith, C. M.; Beale, K. M.; Towle, D. W.; Schooley, D. A.; Dickinson, P. S.

    2010-01-01

    In insects, a family of peptides with sequence homology to the vertebrate calcitonins has been implicated in the control of diuresis, a process that includes mixing of the hemolymph. Here, we show that a member of the insect calcitonin-like diuretic hormone (CLDH) family is present in the American lobster, Homarus americanus, serving, at least in part, as a powerful modulator of cardiac output. Specifically, during an ongoing EST project, a transcript encoding a putative H. americanus CLDH precursor was identified; a full-length cDNA was subsequently cloned. In silico analyses of the deduced prepro-hormone predicted the mature structure of the encoded CLDH to be GLDLGLGRGFSGSQAAKHLMGLAAANFAGGPamide (Homam-CLDH), which is identical to a known Tribolium castaneum peptide. RT-PCR tissue profiling suggests that Homam-CLDH is broadly distributed within the lobster nervous system, including the cardiac ganglion (CG), which controls the movement of the neurogenic heart. RT-PCR analysis conducted on pacemaker neuron- and motor neuron-specific cDNAs suggests that the motor neurons are the source of the CLDH message in the CG. Perfusion of Homam-CLDH through the isolated lobster heart produced dose-dependent increases in both contraction frequency and amplitude and a dose-dependent decrease in contraction duration, with threshold concentrations for all parameters in the range 10–11 to 10–10 mol l–1 or less, among the lowest for any peptide on this system. This report is the first documentation of a decapod CLDH, the first demonstration of CLDH bioactivity outside the Insecta, and the first detection of an intrinsic neuropeptide transcript in the crustacean CG. PMID:20008368

  12. Diversity in the structures and ligand-binding sites of nematode fatty acid and retinol-binding proteins revealed by Na-FAR-1 from Necator americanus.

    PubMed

    Rey-Burusco, M Florencia; Ibáñez-Shimabukuro, Marina; Gabrielsen, Mads; Franchini, Gisela R; Roe, Andrew J; Griffiths, Kate; Zhan, Bin; Cooper, Alan; Kennedy, Malcolm W; Córsico, Betina; Smith, Brian O

    2015-11-01

    Fatty acid and retinol-binding proteins (FARs) comprise a family of unusual α-helix rich lipid-binding proteins found exclusively in nematodes. They are secreted into host tissues by parasites of plants, animals and humans. The structure of a FAR protein from the free-living nematode Caenorhabditis elegans is available, but this protein [C. elegans FAR-7 (Ce-FAR-7)] is from a subfamily of FARs that does not appear to be important at the host/parasite interface. We have therefore examined [Necator americanus FAR-1 (Na-FAR-1)] from the blood-feeding intestinal parasite of humans, N. americanus. The 3D structure of Na-FAR-1 in its ligand-free and ligand-bound forms, determined by NMR (nuclear magnetic resonance) spectroscopy and X-ray crystallography respectively, reveals an α-helical fold similar to Ce-FAR-7, but Na-FAR-1 possesses a larger and more complex internal ligand-binding cavity and an additional C-terminal α-helix. Titration of apo-Na-FAR-1 with oleic acid, analysed by NMR chemical shift perturbation, reveals that at least four distinct protein-ligand complexes can be formed. Na-FAR-1 and possibly other FARs may have a wider repertoire for hydrophobic ligand binding, as confirmed in the present study by our finding that a range of neutral and polar lipids co-purify with the bacterially expressed recombinant protein. Finally, we show by immunohistochemistry that Na-FAR-1 is present in adult worms with a tissue distribution indicative of possible roles in nutrient acquisition by the parasite and in reproduction in the male.

  13. Analysis of immune gene expression modulated by benzo[a]pyrene in head kidney of olive flounder (Paralichthys olivaceus).

    PubMed

    Hur, Deokhwe; Jeon, Joong-Kyun; Hong, Suhee

    2013-05-01

    Poly aromatic hydrocarbons (PAHs) are known to cause functional disorder of fish immune responses. Alteration of inflammatory cytokines and other immune gene expressions by PAHs in immune organs may play a pivotal role in immunotoxicity. Thus this study aimed to elucidate the immunotoxic mechanism of PAH using benzo[a]pyrene (BaP) by analyzing the gene expression of cytokines (IL-1β, TNFα, IL-6, IL-8, IFNγ, Mx), apoptosis (FasL, SOD) and other immune related substances (Lysozyme, IgM) in head kidney and macrophage in olive flounder. In Q-PCR analysis, proinflammatory cytokine (IL-1β, IL-6, IL-8, TNFα) gene expressions were significantly upregulated by BaP while Mx and IgM gene expressions were significantly downregulated in head kidney by a longer exposure to BaP in vivo and in vitro. Lysozyme gene expression was initially upregulated but later downregulated in head kidney in vivo and in vitro. Inhibition test revealed that TNFα gene expression was upregulated by BaP via the AHR pathway as blocked by ANF while IL-6 and IFNγ gene expressions were upregulated by a calcium dependent pathway (i.e. NFAT) as blocked by EGTA. In primary macrophage cells, only IL-8 gene expression was significantly upregulated among proinflammatory cytokines while IFNγ, lysozyme and IgM gene expressions were downregulated by BaP. FasL and SOD expressions were not altered in head kidney cells but significantly upregulated in macrophage cells, indicating apoptosis and oxidative stress. These results indicate that exposure to BaP causes the downregulation of immune response by triggering the death of macrophage cells, the reduction of effectors like IgM and lysozyme, and the decrease of macrophage cell activity.

  14. Recombinant production and evaluation of an antibacterial L-amino acid oxidase derived from flounder Platichthys stellatus.

    PubMed

    Kasai, Kosuke; Hashiguchi, Kenro; Takahashi, Hiroki; Kasai, Ayano; Takeda, Sadanori; Nakano, Manabu; Ishikawa, Takashi; Nakamura, Toshiya; Miura, Tomisato

    2015-08-01

    Fish produce mucus substances as a defensive outer barrier against several bacterial infections. We have recently identified an antibacterial L-amino acid oxidase (psLAAO1) in the mucus layer of the flounder Platichthys stellate. In this study, the antibacterial protein psLAAO1 was expressed as a secretory bioactive recombinant protein in the methylotrophic yeast Pichia pastoris. The recombinant psLAAO1 inhibited the growth of bacteria to the same levels as native psLAAO1 present in mucus. In particular, Staphylococci and Yersinia were strongly suppressed, showing the highest growth retardation of the 21 species and strains tested. Moreover, Staphylococcus epidermidis was most sensitive to psLAAO1 with a minimum inhibitory concentration (MIC) of 0.078 μg/mL, whereas Escherichia coli was essentially resistant to psLAAO1 with a MIC of >10 μg/mL. Interestingly, psLAAO1-treated E. coli were found to upregulate the expression of the btuE gene, which encodes glutathione peroxidase (GPx). The biochemical function of GPx is to reduce free hydrogen peroxide and is induced under response to reactive oxygen species (ROS). Thus, E. coli confers resistance to the reduced free hydrogen peroxide produced by psLAAO1 by increasing GPx levels. Furthermore, the growth of Staphylococcus aureus was completely inhibited in the presence of recombinant psLAAO1. The morphology of psLAAO1-treated S. aureus showed cell surface damage, the formation of large aggregates and the cells showed severe deformations. Western blot analysis showed that psLAAO1 binds to the surface of S. aureus. Therefore, psLAAO1 binds to the surface of LAAO-sensitive S. aureus and directs peroxidative activity at the surface of the bacterial membrane.

  15. Evaluation of cytotoxicity, genotoxicity and embryotoxicity of insecticide propoxur using flounder gill (FG) cells and zebrafish embryos.

    PubMed

    Pandey, Manish Raj; Guo, Huarong

    2014-04-01

    Cytotoxicity, genotoxicity and embryotoxicity of carbamate insecticide propoxur were evaluated using flounder gill (FG) cells and zebrafish embryos. The cytotoxicity of propoxur in FG cells was analyzed by MTT, neutral red uptake (NRU), lactate dehydrogenase (LDH) release and Hoechst 33342 and propidium iodide double staining, and acute cytotoxic effects were observed in a concentration-dependent manner. The 24h-IC50 values of 89.96 ± 1.04, 103.4 ± 1.14 and 86.59 ± 1.13 μg/ml propoxur were obtained by MTT, NRU and LDH assays, respectively. The lethal effects were induced in FG cells mainly through necrosis but not apoptosis as evidenced by double fluorescence staining. Comet assay showed weak genotoxic effects and statistically significant DNA damages were recorded in the cells exposed to highest tested concentration of 75 μg/ml propoxur (p<0.05). Propoxur exerted obvious acute toxic effects on the survival, spontaneous movement, hatching and heart rate, and development (yolk and pericardial sac edema) of zebrafish embryos in both time- and concentration-dependent manner only at ⩾ 100 μg/ml. The corresponding 24h-, 48 h- and 96 h-LC50 values of propoxur in zebrafish embryos were 166.4 ± 1.06, 146.3 ± 1.07 and 134.8 ± 1.06 μg/ml, respectively. The above data obtained suggest a low acute toxicity of propoxur to the in vitro cultured FG cells and zebrafish embryos.

  16. Developmental changes in melanophores and their asymmetrical responsiveness to melanin-concentrating hormone during metamorphosis in barfin flounder (Verasper moseri).

    PubMed

    Yoshikawa, Naoki; Matsuda, Taihei; Takahashi, Akiyoshi; Tagawa, Masatomo

    2013-12-01

    Barfin flounder larvae exhibit unique black coloration, as well as left-right asymmetry in juvenile stage as in other flatfish. In this study, we first assessed the changes in melanophores with development and then investigated their responsiveness to melanin-concentrating hormone (MCH) during metamorphosis. Larval-type melanophores appeared on both sides of the body before metamorphosis, whereas adult-type melanophores appeared only on the ocular side after metamorphosis. Even in the individuals of this species displaying black coloration, the density of larval-type melanophores was similar to that in transparent larvae of other species. However, unlike in transparent larvae, larval-type melanophores completely dispersed in the black larvae of this species. Therefore, the black coloration during larval stages was mainly due to dispersion, and not the density, of larval-type melanophores. In vitro MCH treatment revealed, for the first time, the responsiveness of melanophores in larval stages. On the ocular side, larval-type melanophores aggregated against MCH during larval stages, while, in the larvae at later metamorphic stages and in juveniles, larval-type melanophores did not aggregate, although aggregation of adult-type melanophores was noted. In contrast, on the blind side, the responsiveness of larval-type melanophores to MCH was consistently present from larval to juvenile stages. The metamorphic transition of MCH responsiveness from larval- to adult-type melanophores only on the ocular side suggests the larval (therefore, immature) nature of the blind side skin. We propose that the inhibited development, and thus the retention of the larval-type skin leads to the formation of the blind side characteristics and is the central mechanism for the flatfish asymmetry.

  17. Spatial and annual variation in fecundity and oocyte atresia of yellowtail flounder, Limanda ferruginea, in U.S. waters

    NASA Astrophysics Data System (ADS)

    McElroy, W. David; Wuenschel, Mark J.; Towle, Emilee K.; McBride, Richard S.

    2016-01-01

    Potential annual fecundity (PAF) was estimated over three years (2010-2012) for yellowtail flounder with individuals from the three stocks off the northeast U.S. coast. Down-regulation of PAF, the resorption of oocytes during development, was evident as the vitellogenic cohort advanced, so we directly measured atresia of vitellogenic oocytes using stereological techniques. PAF models including relative fish condition, stock area, year, and oocyte diameter of the leading cohort explained more variation than models with just size alone based on Akaike information criteria. In a given year, Gulf of Maine females had lower PAF at size than southern New England females. Interannual differences were evident: PAF of both stocks was higher in 2010 and lower in 2012, with 2011 showing less synchronization between these stocks. Differences in size at age and relative condition suggested that energy available for somatic and reproductive growth was lower in some years in the Gulf of Maine and Georges Bank, especially in 2011. Georges Bank PAF and condition were intermediate to the other stocks or more similar to the Gulf of Maine, varying annually. A latitudinal gradient in PAF is evident based on our results and relative to earlier studies that included Canadian stocks. The magnitude of down-regulation was variable across stocks and typically 3-25% of PAF. This can be accounted for in fecundity estimates, by the seasonal schedule of sampling and use of an oocyte diameter term in the fecundity model. Theoretical models of atresia patterns suggested variable rates over the later portion of clutch development. The timing of down-regulation varied among years, and its intensity was influenced by female relative condition. Fecundity was related to fish size, but was also affected by fish condition and oocyte diameter (a proxy for time until spawning), and spatial and temporal effects. A longer time series of PAF may identify environmental drivers that modulate annual stock

  18. Evaluation of cytotoxicity and genotoxicity of insecticide carbaryl to flounder gill cells and its teratogenicity to zebrafish embryos

    NASA Astrophysics Data System (ADS)

    Pandey, Manish Raj; Guo, Huarong

    2015-04-01

    In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill (FG) cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was determined with methods including MTT and neutral red uptaking (NRU), lactate dehydrogenase (LDH) releasing and Hoechst 33342 and propidium idodide (PI) double staining. Moderate cytotoxicity in a concentration-dependent manner was observed. The 24 h-IC50 value of 53.48 ± 1.21, 59.13 ± 1.19 and 46.21 ± 1.24 mg L-1 carbaryl was obtained through MTT, NRU and LDH assays, respectively. Double fluorescence staining demonstrated that carbaryl induced the death of FG cells mainly through necrosis. There was no significant genotoxicity found in the FG cells exposed to the highest testing concentration of carbaryl (20 mg L-1, P > 0.05) as was demonstrated by Comet assay. Zebrafish embryos exposed to carbaryl at concentrations ≥10 mg L-1 displayed moderate toxic effects on the survival, spontaneous movement, hatching, heart rates of the embryos and their development, which were evidenced by yolk and pericardial sac edemas, body length reduction and tail flexure in time- and concentration-dependent manners at specific stages. The 24 h-, 48 h- and 96 h-LC50 values of carbaryl to zebrafish embryos were 41.80 ± 1.10, 17.80 ± 1.04 and 14.46 ± 1.05 mg L-1, respectively. These results suggested that carbaryl is moderately toxic to FG cells cultured in vitro and zebrafish embryos, and the FG cells were similar to zebrafish embryos in their sensitivity to carbaryl as 24 h-IC50 and LC50 indicated.

  19. Recombinant production and evaluation of an antibacterial L-amino acid oxidase derived from flounder Platichthys stellatus.

    PubMed

    Kasai, Kosuke; Hashiguchi, Kenro; Takahashi, Hiroki; Kasai, Ayano; Takeda, Sadanori; Nakano, Manabu; Ishikawa, Takashi; Nakamura, Toshiya; Miura, Tomisato

    2015-08-01

    Fish produce mucus substances as a defensive outer barrier against several bacterial infections. We have recently identified an antibacterial L-amino acid oxidase (psLAAO1) in the mucus layer of the flounder Platichthys stellate. In this study, the antibacterial protein psLAAO1 was expressed as a secretory bioactive recombinant protein in the methylotrophic yeast Pichia pastoris. The recombinant psLAAO1 inhibited the growth of bacteria to the same levels as native psLAAO1 present in mucus. In particular, Staphylococci and Yersinia were strongly suppressed, showing the highest growth retardation of the 21 species and strains tested. Moreover, Staphylococcus epidermidis was most sensitive to psLAAO1 with a minimum inhibitory concentration (MIC) of 0.078 μg/mL, whereas Escherichia coli was essentially resistant to psLAAO1 with a MIC of >10 μg/mL. Interestingly, psLAAO1-treated E. coli were found to upregulate the expression of the btuE gene, which encodes glutathione peroxidase (GPx). The biochemical function of GPx is to reduce free hydrogen peroxide and is induced under response to reactive oxygen species (ROS). Thus, E. coli confers resistance to the reduced free hydrogen peroxide produced by psLAAO1 by increasing GPx levels. Furthermore, the growth of Staphylococcus aureus was completely inhibited in the presence of recombinant psLAAO1. The morphology of psLAAO1-treated S. aureus showed cell surface damage, the formation of large aggregates and the cells showed severe deformations. Western blot analysis showed that psLAAO1 binds to the surface of S. aureus. Therefore, psLAAO1 binds to the surface of LAAO-sensitive S. aureus and directs peroxidative activity at the surface of the bacterial membrane. PMID:25661816

  20. EFFECT OF 17B-ESTRADIOL, O,P'-DDT, OCTYLPHENOL AND P,P'-DDE ON GONADAL DEVELOPMENT AND LIVER AND KIDNEY PATHOLOGY IN JUVENILE MALE SUMMER FLOUNDER (PARALICHTYS DENTATUS)

    EPA Science Inventory

    The intent of this study was to compare histopathologically the effect of 17 -estradiol (E2), o,p' DDT, octylphenol and p,p' DDE on gonadal development and liver and kidney condition in sexually immature (juvenile) summer flounder (Paralichthys dentatus). The dorsal sinus of 2-...

  1. EFFECT OF ESTROGENIC (O,P'-DDT; OCTOPHENOL AND ANTI-ANDROGENIC (P'P'-DDE) CHEMICALS ON INDICATORS OF ENDOCRINE STATUS IN JUVENILE MALE SUMMER FLOUNDER (PARALICHTHYS DENTATUS)

    EPA Science Inventory

    Laboratory experiments were conducted with male summer flounder to assess the value of selected measures of endocrine status in fish as indicators of exposure to endocrine-disrupting contaminants. Efficts of 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethane
    (o,p'-D...

  2. Molecular cloning and expression of PoIR2, a novel gene involved in immune response in Japanese flounder ( Paralichthys olivaceus)

    NASA Astrophysics Data System (ADS)

    Li, Shuo; Li, Chunmei; Wang, Xubo; Wang, Yanan; Liu, Zhipeng; Zhai, Teng; Zhang, Quanqi

    2010-03-01

    A novel immune-related gene was expressed in Japanese flounder ( Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 ( P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed as soluble type.

  3. Histological and enzymatic responses of Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × P. dentatus ♂) to chronic heat stress.

    PubMed

    Liu, Yifan; Ma, Daoyuan; Zhao, Chunyan; Wang, Wenqi; Zhang, Xuelei; Liu, Xiao; Liu, Ying; Xiao, Zhizhong; Xu, Shihong; Xiao, Yongshuang; Liu, Qinghua; Li, Jun

    2014-08-01

    This study investigated the effects of long-term heat exposure on Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × summer flounder Paralichthys dentatus ♂). From 24 ± 0.5°C, temperature was increased by 1 ± 0.5°C in a day and was kept at that temperature for 5 days before next rise. Cumulative survival rate (CSR), cumulative survival rate under different temperature (CSR-T), histological alteration, and related enzyme activities were investigated. In P. olivaceus, mass mortality occurred at 29 and 32 °C (the CSR-T dropped to 42.39%), and serious gill damages appeared at 30 and 32°C. Meanwhile, the activities of superoxide dismutase (SOD), catalase (CAT), lysozyme (LZM), and pyruvate kinase (PK) declined around 29 and 32°C (except for CAT). In comparison with P. olivaceus, the CSR of the hybrids was higher, the gill kept a better structural integrity, and the activities of SOD, CAT, LZM, and PK showed tiny fluctuations. The results suggested that during the process of chronic heat stress, P. olivaceus seemed to be more sensitive to 29 and 32°C, and the manifestations in survival, histology, and enzyme activity were generally consistent. For the hybrids, the comparatively insensitivity to high temperature might imply its better heat tolerance.

  4. Histological and enzymatic responses of Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × P. dentatus ♂) to chronic heat stress.

    PubMed

    Liu, Yifan; Ma, Daoyuan; Zhao, Chunyan; Wang, Wenqi; Zhang, Xuelei; Liu, Xiao; Liu, Ying; Xiao, Zhizhong; Xu, Shihong; Xiao, Yongshuang; Liu, Qinghua; Li, Jun

    2014-08-01

    This study investigated the effects of long-term heat exposure on Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × summer flounder Paralichthys dentatus ♂). From 24 ± 0.5°C, temperature was increased by 1 ± 0.5°C in a day and was kept at that temperature for 5 days before next rise. Cumulative survival rate (CSR), cumulative survival rate under different temperature (CSR-T), histological alteration, and related enzyme activities were investigated. In P. olivaceus, mass mortality occurred at 29 and 32 °C (the CSR-T dropped to 42.39%), and serious gill damages appeared at 30 and 32°C. Meanwhile, the activities of superoxide dismutase (SOD), catalase (CAT), lysozyme (LZM), and pyruvate kinase (PK) declined around 29 and 32°C (except for CAT). In comparison with P. olivaceus, the CSR of the hybrids was higher, the gill kept a better structural integrity, and the activities of SOD, CAT, LZM, and PK showed tiny fluctuations. The results suggested that during the process of chronic heat stress, P. olivaceus seemed to be more sensitive to 29 and 32°C, and the manifestations in survival, histology, and enzyme activity were generally consistent. For the hybrids, the comparatively insensitivity to high temperature might imply its better heat tolerance. PMID:24390013

  5. Isolation and characterization of a rhabdovirus from starry flounder (Platichthys stellatus) collected from the northern portion of Puget Sound, Washington, USA

    USGS Publications Warehouse

    Mork, Christina; Hershberger, Paul K.; Kocan, Richard; Batts, William N.; Winton, James R.

    2004-01-01

    The initial characterization of a rhabdovirus isolated from a single, asymptomatic starry flounder (Platichthys stellatus) collected during a viral survey of marine fishes from the northern portion of Puget Sound, Washington, USA, is reported. Virions were bullet-shaped and approximately 100 nm long and 50 nm wide, contained a lipid envelope, remained stable for at least 14 days at temperatures ranging from -80 to 5 degrees C and grew optimally at 15 degrees C in cultures of epithelioma papulosum cyprini (EPC) cells. The cytopathic effect on EPC cell monolayers was characterized by raised foci containing rounded masses of cells. Pyknotic and dark-staining nuclei that also showed signs of karyorrhexis were observed following haematoxylin and eosin, May-Grunwald Giemsa and acridine orange staining. PAGE of the structural proteins and PCR assays using primers specific for other known fish rhabdoviruses, including Infectious hematopoietic necrosis virus, Viral hemorrhagic septicemia virus, Spring viremia of carp virus, and Hirame rhabdovirus, indicated that the new virus, tentatively termed starry flounder rhabdovirus (SFRV), was previously undescribed in marine fishes from this region. In addition, sequence analysis of 2678 nt of the amino portion of the viral polymerase gene indicated that SFRV was genetically distinct from other members of the family Rhabdoviridae for which sequence data are available. Detection of this virus during a limited viral survey of wild fishes emphasizes the void of knowledge regarding the diversity of viruses that naturally infect marine fish species in the North Pacific Ocean.

  6. Isolation and characterization of a rhabdovirus from starry flounder (Platichthys stellatus) collected from the northern portion of Puget Sound, Washington, USA.

    PubMed

    Mork, Christina; Hershberger, Paul; Kocan, Richard; Batts, William; Winton, James

    2004-02-01

    The initial characterization of a rhabdovirus isolated from a single, asymptomatic starry flounder (Platichthys stellatus) collected during a viral survey of marine fishes from the northern portion of Puget Sound, Washington, USA, is reported. Virions were bullet-shaped and approximately 100 nm long and 50 nm wide, contained a lipid envelope, remained stable for at least 14 days at temperatures ranging from -80 to 5 degrees C and grew optimally at 15 degrees C in cultures of epithelioma papulosum cyprini (EPC) cells. The cytopathic effect on EPC cell monolayers was characterized by raised foci containing rounded masses of cells. Pyknotic and dark-staining nuclei that also showed signs of karyorrhexis were observed following haematoxylin and eosin, May-Grunwald Giemsa and acridine orange staining. PAGE of the structural proteins and PCR assays using primers specific for other known fish rhabdoviruses, including Infectious hematopoietic necrosis virus, Viral hemorrhagic septicemia virus, Spring viremia of carp virus, and Hirame rhabdovirus, indicated that the new virus, tentatively termed starry flounder rhabdovirus (SFRV), was previously undescribed in marine fishes from this region. In addition, sequence analysis of 2678 nt of the amino portion of the viral polymerase gene indicated that SFRV was genetically distinct from other members of the family Rhabdoviridae for which sequence data are available. Detection of this virus during a limited viral survey of wild fishes emphasizes the void of knowledge regarding the diversity of viruses that naturally infect marine fish species in the North Pacific Ocean.

  7. Dietary vitamin A, ascorbic acid and α-tocopherol affect the gonad development and reproductive performance of starry flounder Platichthys stellatus broodstock

    NASA Astrophysics Data System (ADS)

    Wang, Jiying; Li, Baoshan; Liu, Xudong; Ma, Jingjing; Wang, Shixin; Zhang, Limin

    2014-03-01

    The present trial was conducted with starry flounder Platichthys stellatus broodstock to evaluate the effects of dietary vitamin A, ascorbic acid, and α-tocopherol on the gonadal development and reproductive performance. 8 000 IU/kg diet vitamin A (VA group), 500 mg/kg diet ascorbic acid (Vcpp group), or 250 mg/kg diet α-tocopherol (α-TA group) was added into basal diet to create 3 vitamin experimental diets, respectively. Each diet was fed to 450 starry flounder broodstock for 104 days. Samples were collected weekly. The gonadosomatic index (GSI) of 3 vitamin experimental groups first decreased and then increased. Maximum GSI of Vcpp group was higher than that of α-TA group but lower than that of VA group. The spawning periods of 3 vitamin experimental groups lasted 49, 56, and 45 days, respectively. No mature eggs were observed in the control group during the trial. The absolute fecundity (AF) and relative fecundity (RF) of α-TA group was higher than that of Vcpp group but lower than that of VA group. The results suggest that different vitamins play different roles in the fish reproductive process. Vitamin A stimulated the maturation of the ovary, ascorbic acid prolonged the spawning period, and α-tocopherol affected the development of the eggs.

  8. Antifreeze protein modulates cell survival during cryopreservation: mediation through influence on ice crystal growth.

    PubMed Central

    Carpenter, J F; Hansen, T N

    1992-01-01

    Antifreeze proteins (AFPs) are extremely efficient at inhibiting ice recrystallization in frozen solutions. Knight and Duman [Knight, C. A. & Duman, J. G. (1986) Cryobiology 23, 256-263] have proposed that this may be an important function of the proteins in freeze-tolerant organisms. We have tested this proposal in vitro by characterizing the influence of AFP on the recovery of cryopreserved cells, which often can survive cooling and yet subsequently be damaged by ice crystal growth during warming. Relatively low concentrations (e.g., 5-150 micrograms/ml) of winter flounder (Pseudopleuronectes americanus) AFP enhance survival of red blood cells cryopreserved in hydroxyethyl starch solutions. This effect is most apparent in samples warmed at suboptimal rates, i.e., where ice recrystallization would be exaggerated. Cryomicroscopy demonstrates that AFP inhibits ice recrystallization in the extracellular regions during the latter stages of the warming cycle. AFP concentrations that enhance survival of red cells confer partial inhibition of recrystallization. Relatively high concentrations of AFP (e.g., 1.54 mg/ml) are much more effective at inhibiting extracellular recrystallization. However, extensive growth of ice around the cell, and concomitant cell damage, is noted. The mechanism for this AFP-induced ice growth is unknown. We propose that there is a delicate balance between AFP-induced enhancement of cell preservation and AFP-induced enhancement of cell preservation and AFP-induced enhancement of cell damage and that this balance hinges on the degrees of inhibition of ice recrystallization and of preferential growth of ice around the cells. We conclude that, under appropriate conditions, one of the proposed functions of AFPs in nature can be emulated, and perhaps have application, in cryopreservation of materials of biomedical interest. Images PMID:1409591

  9. Thermoprotection of a functional epithelium: heat stress effects on transepithelial transport by flounder renal tubule in primary monolayer culture.

    PubMed Central

    Brown, M A; Upender, R P; Hightower, L E; Renfro, J L

    1992-01-01

    Primary monolayer cultures of winter flounder renal proximal-tubule cells were used to determine whether transepithelial transport could be protected from the damaging effects of extreme temperature by previous mild heat shock. Renal tubule epithelial cells were enzymatically dispersed and reorganized as confluent monolayer sheets on native rat tail collagen. Transepithelial electrical properties (potential difference, resistance, short-circuit current, and Na(+)-dependent glucose current) and unidirectional [35S]sulfate fluxes were measured in Ussing chambers at 22 degrees C. Examination of transepithelial electrical properties following acute 1-hr elevation of temperature over a range of 22-37 degrees C provided the basis for the "mild" versus "severe" thermal stress protocols. Severe elevation from 22 degrees C to 32 degrees C for 1.5 hr followed by 1.5 hr at 22 degrees C significantly decreased glucose current (7 +/- 0.7 to 3 +/- 0.8 microA/cm2) as well as net sulfate secretion [131 +/- 11 to 33 +/- 11 nmol/(cm2.hr)]. Mild heat shock of 27 degrees C for 6 hr prior to this severe heat shock completely protected both glucose transport (6 +/- 0.7 microA/cm2) and sulfate flux (149 +/- 13 nmol/(cm2.hr)]. Scanning electron microscopy showed that the number of microvilli on the apical (luminal) surface of the epithelium was decreased after a 32 degrees C heat shock. Monolayers exposed to 27 degrees C for 6 hr prior to incubation at 32 degrees C showed no loss of microvilli. SDS/PAGE analysis of protein patterns from the cultures showed that three classes of heat shock proteins were maximally induced at 27 degrees C. Inhibition of protein synthesis by cycloheximide prevented the thermoprotective effect of mild heat shock. This suggests that certain renal transport functions can be protected from sublethal but debilitating thermal stress by prior mild heat shock and that heat shock proteins may play a role in this protection. Images PMID:1565616

  10. Optimization and revision of the production process of the Necator americanus glutathione S-transferase 1 (Na-GST-1), the lead hookworm vaccine recombinant protein candidate.

    PubMed

    Curti, Elena; Seid, Christopher A; Hudspeth, Elissa; Center, Lori; Rezende, Wanderson; Pollet, Jeroen; Kwityn, Cliff; Hammond, Molly; Matsunami, Rise K; Engler, David A; Hotez, Peter J; Elena Bottazzi, Maria

    2014-01-01

    Infection by the human hookworm Necator americanus is a leading cause of anemia and disability in the developing countries of Africa, Asia, and the Americas. In order to prevent childhood hookworm disease in resource poor settings, a recombinant vaccine is under development by the Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, a Product Development Partnership (PDP). Previously, we reported on the expression and purification of a highly promising hookworm vaccine candidate, Na-GST-1, an N. americanus glutathione s-transferase expressed in Pichia pastoris (yeast), which led to production of 1.5 g of 95% pure recombinant protein at a 20L scale. (1) (,) (2) (,) (3) This yield and purity of Na-GST-1 was sufficient for early pilot manufacturing and initial phase 1 clinical testing. However, based on the number of doses which would be required to allow mass vaccination and a potential goal to deliver a vaccine as inexpensively as possible, a higher yield of expression of the recombinant antigen at the lowest possible cost is highly desirable. Here we report on modifications to the fermentation (upstream process) of the antigen expressed in P. pastoris, and to the purification (downstream process) of the recombinant protein that allowed for a 2-3-fold improvement in the final yield of Na-GST-1 purified protein. The major improvements included upstream process changes such as the addition of a sorbitol pulse and co-feed during methanol induction as well as an extension of the induction stage to approximately 96 hours; downstream process changes included modifying the UFDF to flat sheet with a 10 kDa Molecular Weight cut-off (MWCO), adjusting the capacity of an ion-exchange chromatography step utilizing a gradient elution as opposed to the original step elution, and altering the hydrophobic interaction chromatography conditions. The full process, as well as the purity and stability profiles of the target Na-GST-1, and its formulation

  11. A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone on the induction of spermiation and amplexus in the American toad (Anaxyrus americanus)

    PubMed Central

    2012-01-01

    Background Captive breeding programs for endangered amphibian species often utilize exogenous hormones for species that are difficult to breed. The purpose of our study was to compare the efficacy of two different hormones at various concentrations on sperm production, quantity and quality over time in order to optimize assisted breeding. Methods Male American toads (Anaxyrus americanus) were divided into three separate treatment groups, with animals in each group rotated through different concentrations of luteinizing hormone releasing hormone analog (LHRH; 0.1, 1.0, 4.0 and 32 micrograms/toad), human chorionic gonadotropin (hCG; 50, 100, 200, and 300 IU), or the control over 24 hours. We evaluated the number of males that respond by producing spermic urine, the sperm concentration, percent motility, and quality of forward progression. We also evaluated the effects of hCG and LHRH on reproductive behavior as assessed by amplexus. Data were analyzed using the Generalized Estimating Equations incorporating repeated measures over time and including the main effects of treatment and time, and the treatment by time interaction. Results The hormone hCG was significantly more effective at stimulating spermiation in male Anaxyrus americanus than LHRH and showed a dose-dependent response in the number of animals producing sperm. At the most effective hCG dose (300 IU), 100% of the male toads produced sperm, compared to only 35% for the best LHRH dose tested (4.0 micrograms). In addition to having a greater number of responders (P < 0.05), the 300 IU hCG treatment group had a much higher average sperm concentration (P < 0.05) than the treatment group receiving 4.0 micrograms LHRH. In contrast, these two treatments did not result in significant differences in sperm motility or quality of forward progressive motility. However, more males went into amplexus when treated with LHRH vs. hCG (90% vs. 75%) by nine hours post-administration. Conclusion There is a clear

  12. Insulin-like growth factors I and II in starry flounder (Platichthys stellatus): molecular cloning and differential expression during embryonic development.

    PubMed

    Xu, Yongjiang; Zang, Kun; Liu, Xuezhou; Shi, Bao; Li, Cunyu; Shi, Xueying

    2015-02-01

    In order to elucidate the possible roles of insulin-like growth factors I and II (IGF-I and IGF-II) in the embryonic development of Platichthys stellatus, their cDNAs were isolated and their spatial expression pattern in adult organs and temporal expression pattern throughout embryonic development were examined by quantitative real-time PCR assay. The IGF-I cDNA sequence was 1,268 bp in length and contained an open reading frame (ORF) of 558 bp, which encoded 185 amino acid residues. With respect to IGF-II, the full-length cDNA was 899 bp in length and contained a 648-bp ORF, which encoded 215 amino acid residues. The amino acid sequences of IGF-I and IGF-II exhibited high identities with their fish counterparts. The highest IGF-I mRNA level was found in the liver for both sexes, whereas the IGF-II gene was most abundantly expressed in female liver and male liver, gill, and brain. The sex-specific and spatial expression patterns of IGF-I and IGF-II mRNAs are thought to be related to the sexually dimorphic growth and development of starry flounder. Both IGF-I and IGF-II mRNAs were detected in unfertilized eggs, which indicated that IGF-I and IGF-II were parentally transmitted. Nineteen embryonic development stages were tested. IGF-I mRNA level remained high from unfertilized eggs to low blastula followed by a significant decrease at early gastrula and then maintained a lower level. In contrast, IGF-II mRNA level was low from unfertilized eggs to high blastula and peaked at low blastula followed by a gradual decrease. Moreover, higher levels of IGF-I mRNA than that of IGF-II were found from unfertilized eggs to high blastula, vice versa from low blastula to newly hatched larva, and the different expression pattern verified the differential roles of IGF-I and IGF-II in starry flounder embryonic development. These results could help in understanding the endocrine mechanism involved in the early development and growth of starry flounder.

  13. Insulin-like growth factors I and II in starry flounder (Platichthys stellatus): molecular cloning and differential expression during embryonic development.

    PubMed

    Xu, Yongjiang; Zang, Kun; Liu, Xuezhou; Shi, Bao; Li, Cunyu; Shi, Xueying

    2015-02-01

    In order to elucidate the possible roles of insulin-like growth factors I and II (IGF-I and IGF-II) in the embryonic development of Platichthys stellatus, their cDNAs were isolated and their spatial expression pattern in adult organs and temporal expression pattern throughout embryonic development were examined by quantitative real-time PCR assay. The IGF-I cDNA sequence was 1,268 bp in length and contained an open reading frame (ORF) of 558 bp, which encoded 185 amino acid residues. With respect to IGF-II, the full-length cDNA was 899 bp in length and contained a 648-bp ORF, which encoded 215 amino acid residues. The amino acid sequences of IGF-I and IGF-II exhibited high identities with their fish counterparts. The highest IGF-I mRNA level was found in the liver for both sexes, whereas the IGF-II gene was most abundantly expressed in female liver and male liver, gill, and brain. The sex-specific and spatial expression patterns of IGF-I and IGF-II mRNAs are thought to be related to the sexually dimorphic growth and development of starry flounder. Both IGF-I and IGF-II mRNAs were detected in unfertilized eggs, which indicated that IGF-I and IGF-II were parentally transmitted. Nineteen embryonic development stages were tested. IGF-I mRNA level remained high from unfertilized eggs to low blastula followed by a significant decrease at early gastrula and then maintained a lower level. In contrast, IGF-II mRNA level was low from unfertilized eggs to high blastula and peaked at low blastula followed by a gradual decrease. Moreover, higher levels of IGF-I mRNA than that of IGF-II were found from unfertilized eggs to high blastula, vice versa from low blastula to newly hatched larva, and the different expression pattern verified the differential roles of IGF-I and IGF-II in starry flounder embryonic development. These results could help in understanding the endocrine mechanism involved in the early development and growth of starry flounder. PMID:25424555

  14. Metabolic responses to hypoxia of Lycenchelys verrillii (wolf eelpout) and Glyptocephalus cynoglossus (witch flounder): Sedentary bottom fishes of the Hatteras/Virginia middle slope

    USGS Publications Warehouse

    Moser, M.L.; Ross, S.W.; Sulak, K.J.

    1996-01-01

    We collected wolf eelpouts Lycenchelys verrillii and witch flounder Glyptocephalus cynoglossus from Hatteras (North Carolina, USA) and Virginia (USA) Middle Slope sites using a submersible, and made shipboard measurements of their respiration rates and survival in hypoxic (<10% O2 saturation) and anoxic conditions. Both species from the Hatteras site reduced their respiration rates as ambient oxygen decreased, but eelpouts from the Virginia site maintained a constant respiration rate until oxygen saturation dropped below 20%. Moreover, eelpouts from the Hatteras site were significantly more tolerant of hypoxic conditions than fish from the Virginia site and survived anoxia for short periods. These results and our submersible observations of fish behavior support the hypothesis that the Hatteras Middle Slope fauna is exposed to short-term hypoxia events.

  15. Incidence of Infestation and Larval Success of Emerald Ash Borer (Agrilus planipennis) on White Fringetree (Chionanthus virginicus), Chinese Fringetree (Chionanthus retusus), and Devilwood (Osmanthus americanus).

    PubMed

    Cipollini, Don; Rigsby, Chad M

    2015-10-01

    We compared the incidence of infestation by emerald ash borer (EAB) and lilac borer on white fringetree to that of its Asian congener, Chinese fringetree, Chionanthus retusus, and a North American relative, devilwood, Osmanthus americanus. We also conducted laboratory bioassays to determine the suitability of these hosts for EAB larvae. At Spring Grove Cemetery and Arboretum in Cincinnati, Ohio, 9 of 28 white fringetrees examined were infested by EAB. Most of the white fringetrees had lilac borer infestation, and most of the trees infested by EAB also had lilac borer infestation. None of the 11 Chinese fringetrees examined were infested by either EAB or lilac borer. Each of the five devilwood individuals examined was infested by lilac borer, but not EAB. At The Morton Arboretum in Lisle, Illinois, 7 of 16 white fringetrees examined were infested by EAB, while none of the seven Chinese fringetrees examined were infested by either insect. A 40-d bioassay confirmed that white fringetree was an acceptable host, producing fourth-instar larvae that were smaller than those produced on a highly susceptible cultivar of green ash, Fraxinus pennsylvanica. No larvae survived on Chinese fringetree, and neonates were largely incapable of feeding on it. Two larvae survived on devilwood, reaching the second instar and excavating extensive galleries. Future work should be aimed at biotic and abiotic factors influencing the susceptibility of white fringetree, as well as further examination of close relatives for their vulnerability to EAB. PMID:26314014

  16. Molecular phylogeny and SNP variation of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) derived from genome sequences.

    PubMed

    Cronin, Matthew A; Rincon, Gonzalo; Meredith, Robert W; MacNeil, Michael D; Islas-Trejo, Alma; Cánovas, Angela; Medrano, Juan F

    2014-01-01

    We assessed the relationships of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) with high throughput genomic sequencing data with an average coverage of 25× for each species. A total of 1.4 billion 100-bp paired-end reads were assembled using the polar bear and annotated giant panda (Ailuropoda melanoleuca) genome sequences as references. We identified 13.8 million single nucleotide polymorphisms (SNP) in the 3 species aligned to the polar bear genome. These data indicate that polar bears and brown bears share more SNP with each other than either does with black bears. Concatenation and coalescence-based analysis of consensus sequences of approximately 1 million base pairs of ultraconserved elements in the nuclear genome resulted in a phylogeny with black bears as the sister group to brown and polar bears, and all brown bears are in a separate clade from polar bears. Genotypes for 162 SNP loci of 336 bears from Alaska and Montana showed that the species are genetically differentiated and there is geographic population structure of brown and black bears but not polar bears.

  17. Application of Large-Scale Parentage Analysis for Investigating Natal Dispersal in Highly Vagile Vertebrates: A Case Study of American Black Bears (Ursus americanus)

    PubMed Central

    Moore, Jennifer A.; Draheim, Hope M.; Etter, Dwayne; Winterstein, Scott; Scribner, Kim T.

    2014-01-01

    Understanding the factors that affect dispersal is a fundamental question in ecology and conservation biology, particularly as populations are faced with increasing anthropogenic impacts. Here we collected georeferenced genetic samples (n = 2,540) from three generations of black bears (Ursus americanus) harvested in a large (47,739 km2), geographically isolated population and used parentage analysis to identify mother-offspring dyads (n = 337). We quantified the effects of sex, age, habitat type and suitability, and local harvest density at the natal and settlement sites on the probability of natal dispersal, and on dispersal distances. Dispersal was male-biased (76% of males dispersed) but a small proportion (21%) of females also dispersed, and female dispersal distances (mean ± SE  =  48.9±7.7 km) were comparable to male dispersal distances (59.0±3.2 km). Dispersal probabilities and dispersal distances were greatest for bears in areas with high habitat suitability and low harvest density. The inverse relationship between dispersal and harvest density in black bears suggests that 1) intensive harvest promotes restricted dispersal, or 2) high black bear population density decreases the propensity to disperse. Multigenerational genetic data collected over large landscape scales can be a powerful means of characterizing dispersal patterns and causal associations with demographic and landscape features in wild populations of elusive and wide-ranging species. PMID:24621593

  18. Molecular phylogeny and SNP variation of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) derived from genome sequences.

    PubMed

    Cronin, Matthew A; Rincon, Gonzalo; Meredith, Robert W; MacNeil, Michael D; Islas-Trejo, Alma; Cánovas, Angela; Medrano, Juan F

    2014-01-01

    We assessed the relationships of polar bears (Ursus maritimus), brown bears (U. arctos), and black bears (U. americanus) with high throughput genomic sequencing data with an average coverage of 25× for each species. A total of 1.4 billion 100-bp paired-end reads were assembled using the polar bear and annotated giant panda (Ailuropoda melanoleuca) genome sequences as references. We identified 13.8 million single nucleotide polymorphisms (SNP) in the 3 species aligned to the polar bear genome. These data indicate that polar bears and brown bears share more SNP with each other than either does with black bears. Concatenation and coalescence-based analysis of consensus sequences of approximately 1 million base pairs of ultraconserved elements in the nuclear genome resulted in a phylogeny with black bears as the sister group to brown and polar bears, and all brown bears are in a separate clade from polar bears. Genotypes for 162 SNP loci of 336 bears from Alaska and Montana showed that the species are genetically differentiated and there is geographic population structure of brown and black bears but not polar bears. PMID:24477675

  19. Effects of molting and environmental factors on trace metal body-burdens and hemocyanin concentrations in the American lobster, Homarus americanus.

    PubMed

    Engel, D W; Brouwer, M; Mercaldo-Allen, R

    2001-09-01

    Hemocyanin concentrations in the hemolymph of marine crustacea are dependent on the molt cycle and on environmental conditions. Studies in our laboratories have found that hemocyanin levels in blue crabs are reduced after ecdysis and under conditions of environmental stress (Engel, Brouwer, & McKenna, 1993. Hemocyanin concentrations in marine crustaceans as a function of environmental conditions. Marine Ecology Progress Series, 93, 233-244). We have extended those studies to include the American lobster, Homarus americanus. Hemolymph and digestive gland tissues from Long Island Sound lobsters were analyzed for hemocyanin, copper, and zinc during different stages of the molt cycle. Hemocyanin, copper and zinc in the hemolymph were highest in premolt stages (D1-D4), and lowest in the postecdysal papershell stages (B1-B2). Concomitantly, copper in digestive glands decreased significantly following ecdysis, but no significant changes in the metals bound to metallothionein (MT) were observed. Copper-MT was the predominant form throughout the molt cycle, presumably because lobsters were obtained from copper-contaminated areas. To examine the effects of environmental factors, intermolt lobsters were collected from locations of different environmental quality along the Atlantic coast, and were analyzed for hemocyanin and trace metals. In general, animals from areas with a history of contamination showed the highest hemocyanin concentrations.

  20. Crystallization and preliminary X-ray analysis of Na-ASP-1, a multi-domain pathogenesis-related-1 protein from the human hookworm parasite Necator americanus

    PubMed Central

    Asojo, Oluwatoyin A.; Loukas, Alex; Inan, Mehmet; Barent, Rick; Huang, Jicai; Plantz, Brad; Swanson, Amber; Gouthro, Mark; Meagher, Michael M.; Hotez, Peter J.

    2005-01-01

    Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis-related-1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been expressed, purified and crystallized. Na-ASP-1 is a 406-amino-acid multi-domain ASP from the prevalent human hookworm parasite Necator americanus. Useful X-ray data to 2.2 Å have been collected from a crystal that belongs to the monoclinic space group P21 with unit-cell parameters a = 67.7, b = 74.27, c = 84.60 Å, β = 112.12°. An initial molecular-replacement solution has been obtained with one monomer in the asymmetric unit. PMID:16511050