The cytochrome P450 system of Atlantic salmon (Salmo salar): II. Variations in hepatic catalytic activities and isozyme patterns during an annual reproductive cycle

A group of Atlantic salmon (Salmo salar) was followed through their first year of maturation and spawning. At monthly intervals, starting with juvenile fish in December, 5–7 fish of each sex were killed, and liver and plasma were sampled. The last sampling point was of spawning fish in November a year later. Variables in the cytochrome P450 (P450) system were studied in hepatic microsomes, and estradiol 17 was measured in the plasma of females to assess the maturational status. The P450 1A1-mediated 7-ethoxyresorufin O-deethylase (EROD) started at high levels in winter, but decreased to non-detectable activities in pre-spawning females. Decreases, but not to the same extent, were also observed during this period in total cytochrome P450, cytochrome b₅, NADPH-cytochrome P450 reductase, and in the content of two immunochemically determined P450 isozymes. At the same time, LSI levels increased in maturing females (starting in July), and GSI levels increased in both sexes (starting in May). Sex specific differences were observed in pre-spawning fish in September and October, with levels of total P450, b₅, NADPH-cytochrome P450 reductase, EROD and P450 isozymes significantly lower in females. At the same time, plasma estradiol-17 levels reached peak values in females. The results point to the important role of sex steroids such as estradiol-17 as major factors in the regulation of final sexual maturation. However, this study also indicates that there may be estradiol-17 independent events of equal importance in the early stages of gonadal maturation that may involve the P450 system. The changes observed in the P450 system (as a major drug and steroid metabolizing system) of Atlantic salmon during sexual maturation may be of importance both in the endogenous transduction of hormonal signals, and as a pharmacological basis for designing therapeutic treatment of diseases in the aquaculture industry.Parts of this work were presented at the 5th International Symposium on Responses of Marine Organisms to Pollutants, April 1989 in Plymouth, United Kingdom (Larsen and Goks\/oyr 1989).     

Accurate Dereplication of Bioactive Secondary Metabolites from Marine-Derived Fungi by UHPLC-DAD-QTOFMS and a MS/HRMS Library

In drug discovery, reliable and fast dereplication of known compounds is essential for identification of novel bioactive compounds. Here, we show an integrated approach using ultra-high performance liquid chromatography-diode array detection-quadrupole time of flight mass spectrometry (UHPLC-DAD-QTOFMS) providing both accurate mass full-scan mass spectrometry (MS) and tandem high resolution MS (MS/HRMS) data. The methodology was demonstrated on compounds from bioactive marine-derived strains of Aspergillus, Penicillium, and Emericellopsis, including small polyketides, non-ribosomal peptides, terpenes, and meroterpenoids. The MS/HRMS data were then searched against an in-house MS/HRMS library of ~1300 compounds for unambiguous identification. The full scan MS data was used for dereplication of compounds not in the MS/HRMS library, combined with ultraviolet/visual (UV/Vis) and MS/HRMS data for faster exclusion of database search results. This led to the identification of four novel isomers of the known anticancer compound, asperphenamate. Except for very low intensity peaks, no false negatives were found using the MS/HRMS approach, which proved to be robust against poor data quality caused by system overload or loss of lock-mass. Only for small polyketides, like patulin, were both retention time and UV/Vis spectra necessary for unambiguous identification. For the ophiobolin family with many structurally similar analogues partly co-eluting, the peaks could be assigned correctly by combining MS/HRMS data and m/z of the [M + Na]⁺ ions.     

Macronutrient (N, P, K) and Redoximorphic Metal (Fe, Mn) Allocation in Leersia oryzoides (Rice Cutgrass) Grown Under Different Flood Regimes

Vegetated drainages are an effective method for removal of pollutants associated with agricultural runoff. Leersia oryzoides, a plant common to agricultural ditches, may be particularly effective in remediation; however, research characterizing responses of L. oryzoides to flooding are limited. Soil reduction resulting from flooding can change availability of nutrients to plants via changes in chemical species (e.g., increasing solubility of Fe). Additionally, plant metabolic stresses resulting from reduced soils can decrease nutrient uptake and translocation. The objective of this study was to characterize belowground and aboveground nutrient allocation of L. oryzoides subjected to various soil moisture regimes. Treatments included: a well-watered and well-drained control; a continuously saturated treatment; a 48-h pulse-flood treatment; and a partially flooded treatment in which water level was maintained at 15 cm below the soil surface and flooded to the soil surface for 48 h once a week. Soil redox potential (Eh, mV) was measured periodically over the course of the 8-week experiment. At experiment termination, concentrations of Kjeldahl nitrogen, phosphorus (P), potassium (K), iron (Fe), and manganese (Mn) were measured in plant tissues. All flooded treatments demonstrated moderately reduced soil conditions (Eh < 350 mV). Plant Kjeldahl nitrogen concentrations demonstrated no treatment effect, whereas P and K concentrations decreased in aboveground portions of the plant. Belowground concentrations of P, Mn, and Fe were significantly higher in flooded plants, likely due to the increased solubility of these nutrients resulting from the reductive decomposition of metal–phosphate complexes in the soil and subsequent precipitation in the rhizosphere. These results indicate that wetland plants may indirectly affect P, Mn, and Fe concentrations in surface waters by altering local trends in soil oxidation–reduction chemistry.     

Effects of wind turbines and other physical elements on field utilization by pink-footed geese (Anser brachyrhynchus): A landscape perspective

The effects of wind turbines and other physical landscape elements on field utilization by wintering pink-footed geese (Anser brachyrhynchus) were studied in a Danish farmland landscape. Within the study area geese were feeding on pastures, which together with cereals were the main crop types. Apart from wind turbines a variety of potentially disturbing landscape elements was present, e.g., high-power lines, windbreaks, roads and settlements. Patterns of field use were assessed by measuring goose dropping densities along transects perpendicular to wind farms (with turbines in clusters and in lines) and other landscape elements. Local effects were expressed in terms of `avoidance distance', i.e., the distance from a given landscape element to the point at which 50% of maximal dropping density was reached. The spatial distribution of landscape elements within an eight km radius from the goose roost was determined from aerial photographs. The area occupied by various elements, together with the adjacent zones which were not available to geese due to their associated avoidance distances, were quantified using Geographic Information System (GIS).The avoidance distance of wind farms with turbines in lines and in clusters were ca 100 m and ca 200 m, respectively. Geese did not enter the area between turbines within the cluster. At the landscape level, the combined effect of physical elements other than wind turbines caused an effective loss of 68% of the total field area (40 km²). Wind turbines caused an additional loss of 4% of the field area. However, of the remaining area available to geese (13 km²), wind turbines caused a loss of 13% of the total area. The habitat loss per turbine was higher for the wind farm with turbines arranged in a large cluster than for wind farms with turbines in small clusters or lines. This difference was mainly due to the fact that wind farms in small clusters or with a linear layout were generally placed close to roads or other elements with existing associated avoidance zones, whereas the large cluster was placed in the open farmland area. The avoidance zones associated with physical elements in the landscape do not take into account possible synergistic effects and, hence, actual field areas affected are likely to be minimum estimates. Implications of these findings for planning of wind farms in areas of conservation interest to geese are discussed.     

Monensin controlled-release intraruminal capsule for control of bloat in pastured dairy cows

Monensin, a polyether ionophore antibiotic, is potentially an important agent for bloat relief in dairy cows grazing temperate legume-based pasture. A series of studies was undertaken to determine the effect of monensin, when delivered continuously in the rumen of lactating dairy cows by means of controlled-release capsules (monensin CRC). Such devices release approximately 300 mg/head/day for 100 d. A short-term pilot study made at Ruakura, New Zealand, tested monensin CRC in cows selected for high susceptibility to bloat and grazing lucerne (Medicago sativa) or red clover (Trifolium pratense). Treatment significantly reduced the incidence of bloat, while milk yield and protein yield were increased. There was no effect on fat yield. Following the pilot study, 6 large-scale field experiments involving a total of 368 lactating dairy cows, were made in Australia and New Zealand to confirm the effectiveness of monensin CRC for bloat control and to measure the effect of such treatment on milk production and composition. A severe bloat problem occurred in 2 experiments, mild bloat occurred in 2 others, while no visual signs of bloat were observed in the remaining 2 experiments. Bloat was significantly (P less than 0.05) reduced by monensin CRC treatment when data was pooled over the 4 experiments in which bloat occurred. Daily milk yield was increased in all experiments from a mean of 17.7 in untreated groups to 18.8 kg/head/day (P less than 0.05) in monensin CRC-treated cows. Protein percentage was not affected by treatment, while there was a decrease from 4.29 to 4.10% fat, although total fat yield was not affected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estimating genetic relationships among historical sources of alfalfa germplasm and selected cultivars with sequence related amplified polymorphisms

Fifteen alfalfa populations consisting of six public cultivars and nine historically recognized sources of alfalfa germplasm in North American cultivars were examined using sequence related amplified polymorphisms (SRAPs). Three bulk DNA samples from each population were evaluated with fourteen different SRAP primer pairs. This resulted in 249 different amplicons, of which over 90% were polymorphic. A dendrogram from the analysis suggests that the public cultivars are quite diverse from all the historical sources of germplasm. The highest mean genetic similarity among the nine original sources of Medicago germplasm was 0.85 between PI 536535 (Peruvian) and 536536 (Indian), while the lowest (0.47) was between PI 560333 (M. falcata) and 536539 (African). The highest mean genetic similarity among the nine original sources of Medicago germplasm and the public alfalfa cultivars was 0.78 between PI 536532 (Ladak) and Vernal, while the lowest (0.59) was between PI 536539 (African) and Oneida. Relationships based on SRAP analysis appear to generally concur with expected relationships based on fall dormancy. This report demonstrates that SRAPs are a promising marker system for detecting polymorphisms in alfalfa.     

Monitoring fish stocks in relation to acidification in Norwegian watersheds

The Norwegian Monitoring Programme for Long-Range Transported Air Pollutants started in 1980. The biological part of this programme includes besides invertebrate studies in streams, (i) fish community status in lakes by means of interviews, test-fishing in lakes by using standard gill-net series, recruitment studies of brown trout in inland streams, and juvenile stock assess and monitoring of fish kills in salmon rivers. Damaged fish stocks are recognized within a land area of 51,500 km² in southern Norway and 30 km² in northern Norway. At least 6,000 lake-dwelling fish stocks have either been lost or are at various stages of reduction. Brown trout (Salmo trutta) is the most widespread and abundant species of fish in Norwegian watersheds, and is also most severe affected by acidification. More recently, there are some indications of an increase in the abundance of brown trout in some areas. However, analysis of age structure in lakes, and fry densities in streams in such areas revealed large annual variations in recruitment rate, which indicates unstable water chemical conditions. Atlantic salmon (Salmo salar) is virtually extinct in 25 rivers in southernmost, southwestern and western Norway.     

Fatty acid composition of various ectomycorrhizal fungi and ectomycorrhizas of Norway spruce

Whole cell fatty acid (WCFA) compositions of three different structures of ectomycorrhizal (ECM) fungi: sporocarps, pure culture mycelia and ectomycorrhizas were analysed to evaluate the potential use of fatty acid profiles as biomarkers for ECM fungi and ectomycorrhiza-associated bacteria. Sporocarps of Amanita muscaria, Amanita rubescens, Lactarius rufus, Lactarius thejogalus, Leccinum scabrum, Paxillus involutus, Russula foetens, Russula rosea, Russula vesca, Suillus grevillei, Tylopilus felleus, Xerocomus badius, Xerocomus subtomentosus, pure cultures of A. muscaria, P. involutus, X. badius, X. subtomentosus, Suillus bovinus Suillus luteus and seven ectomycorrhizal morphotypes of Norway spruce were examined. Our results revealed species-specific composition of fatty acids of fungal sporocarps and pure culture mycelia. Ectomycorrhizal morphotypes distinguished and identified by morphological and molecular methods (PCR-RLFP and sequencing) created specific fatty acid profiles. The dominating fatty acids in pure cultures and sporocarps were 18:2ω6,9, 18:1ω9 and 16:0, whereas ectomycorrhizas also contained plant and bacterial specific fatty acids. Especially, fatty acids specific to Gram-positive bacteria 15:0 anteiso and 17:0 anteiso were present in relatively high amounts and suggested that these bacteria are dominating in the examined Norway spruce mycorrhizosphere. In conclusion, our results show that fatty acid based methods can be useful in studies of ectomycorrhizal fungi, both as a quick method for differentiation of fungal species and also in studies of mycorrhiza-associated microorganisms in the field.