The North Sea: source or sink for nitrogen and phosphorus to the Atlantic Ocean?

Annual nitrogen and phosphorus budgets for the whole North Sea taking into account the most recent data available were established. The area considered has a total surface of approximately 700,000km² and corresponds to the definition by OSPARCOM (Oslo and Paris Commission) with the exclusion of the Skagerrak and Kattegat areas. Input and output fluxes were determined at the marine, atmospheric, sediment and continental boundaries, and riverine inputs based on river flows and nutrient concentrations at the river–estuary interface were corrected for possible estuarine retention. The results showed that the North Sea is an extremely complex system subjected to large inter-annual variability of marine water circulation and freshwater land run-off. Consequently, resulting total N (TN) and P (TP) fluxes are extremely variable from 1 year to another and this has an important influence on the budget of these elements. Total inputs to the North Sea are 8870±4860kTNyear^–1 and 494±279kTPyear^–1. Denitrification is responsible for the loss of 23±7% of the TN inputs while sediment burial is responsible for the retention of only of 2±2% of the TP input. For TN, due to the large variability on marine and estuarine fluxes, and to the uncertainty related to the denitrification rate, it was concluded that the North Sea could either be a source (1930kTNyear^–1) or a sink (1700kTNyear^–1) for the waters of the North Atlantic Ocean. For TP it was concluded that the North Sea is mostly a source (–4 to 52kTPyear^–1) for the waters of the North Atlantic Ocean.     

Mode of uptake of sterol by Arthrobacter simplex

The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M _r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10^–3g l^–1h^–1) could not adequately account for the rate of sterol uptake (72×10^–3g l^–1h^–1) and the specific growth rate (56×10^–3 h^–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm^–1), the sterol-grown cells had strong surface activity (40mNm^–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.     

Conservation status of parrot populations in an Atlantic rainforest area of southeastern Brazil

A census of four species of syntopic parrots was carried out using distance sampling methods on São Sebastião island, SE Brazil. Most of the 33593 ha island is covered by mature and secondary Atlantic rainforest. Almost 80% of these forests are within the Ilhabela Park. Although the species counted have marked differences in size and weight, density (individuals/km²) and estimated population size in 23500 ha of well-preserved forests were similar: Amazona farinosa (13.82±5.94; 3247±1395), Pionus maximiliani (15.79±7.04; 3712±1654), Brotogeris tirica (15.05±4.87; 3537±1143) and Pyrrhura frontalis (13.06±5.53; 3068±1298). Encounter rates of Forpus crassirostris and Pionopsitta pileata were very low, which suggests that there is only a small population of these species on the island. The São Sebastião forests still support healthy populations of parrots. Although woodpecker population estimates on the island are large enough to provide nesting sites for parrots, competition for holes with other secondary cavity nesters such as toucans, flycatchers and tytiras, and the selective cutting of dead trees for canoe construction, which is a common practice on the island, may limit hole availability for parrots.     

Increased erythritol production in Torula sp. by Mn2+and Cu2+

The production of erythritol and the erythritol yield from glucose by Torula sp. were improved, in increasing order, by supplementing with 10 mg MnSO₄4H₂O l^–1, 2 mg CuSO₄5H₂O l^–1, and both 10 mg MnSO₄4H₂O l^–1 and 2 mg CuSO₄5H₂O l^–1. Mn²⁺ decreased the intracellular concentration of erythritol, whereas Cu²⁺ increased the activity of erythrose reductase in cells. These results suggest that Mn²⁺ altered the permeability of cells, whereas Cu²⁺ increased the activity of erythrose reductase in cells.     

Sorbitol as a non-repressing carbon source for fed-batch fermentation of recombinant Pichia pastoris

Glycerol/methanol and sorbitol/methanol mixed-feed fermentation strategies for the production of recombinant proteins by Pichia pastoris were compared in order to examine sorbitol's potential as a carbon source. Although P. pastoris does have a lower cell yield on sorbitol than on glycerol, the specific rate of product formation is higher (60 g protein g^–1 dry wth for sorbitol/methanol, vs 45 g protein g^–1 dry wth for glycerol/methanol), resulting in comparable final recombinant expression levels. Importantly, the presence of residual sorbitol in the growth medium appears to be less repressive to the alcohol oxidase promoter in this organism, providing a more forgiving means of operating mixed-feed fed-batch recombinant P. pastoris fermentations.     

Reproduction of the Endangered Killifish Aphanius iberus at Different Salinities

Adult fish of a freshwater population of the Iberian endangered cyprinodontid Aphanius iberus, were induced to reproduce at salinities of 5, 15, 30, 45 and 60ppt. For each salinity five 30l aquaria were used, each one including a male and two females. Maturity and spawning outside the natural season, were obtained at conditions of 22 to 28°C and a photoperiod of 14L:10D. The larvae were fed with rotifers Brachionus plicatilis and Synchaeta cecilia valentina. Experiment lasted 40 days. The first spawning occurred on the 17th day at 45ppt of salinity and the first embryos hatched on the 34th day at 5 and 15ppt salinity. The final average number of larvae per aquarium ranged from 5.2 (45ppt salinity) to 10.8 (15ppt salinity). No significant differences were found between the average values at different salinities (p<0.01).     

Diel Movements of Bat Rays, Myliobatis californica, in Tomales Bay, California: Evidence for Behavioral Thermoregulation?

We used ultrasonic telemetry to examine movement patterns of 11 bat rays, Myliobatis californica, in Tomales Bay, California. Tomales Bay is long (20km) and narrow (1.4km), and is hydrographically separated into outer and inner bay regions. The outer bay (the outermost 8km) is characterized by oceanic conditions while the shallow inner bay (the innermost 12km) features wide seasonal temperature shifts. Five rays were tracked monthly from October 1990 to November 1991 and six rays (four of which carried temperature-sensing transmitters) were tracked daily from 30 June to 16 July 1992. Mean bat ray movement rate was 8.84mmin^–1 (range 4.49 to 13.40mmin^–1) and was not significantly affected by size (p=0.592), tidal stage (p=0.610), or time of day (p=0.327). Movement direction was unrelated to tidal stage (p=0.472) but showed a highly significant diel pattern (p<0.001). From 2:50–14:50h, rays moved toward the warmer and shallower inner bay, while from 14:50–2:50h they moved toward the cooler and deeper outer bay. These telemetry data, along with known bat ray foraging patterns and respiratory temperature-sensitivity, argue for behavioral thermoregulation as the primary influence on this movement pattern.     

Gross nitrogen transformations in soils from uncut and cut boreal upland and peatland coniferous forest stands

Gross and net nitrogen (N) ammonification and nitrification were measured in soils from an uncut and recently cut upland and peatland conifer stand in northwestern Ontario, Canada. Rates of gross total inorganic N immobilization were similar to gross mineralization, resulting in low net mineralization rates in soils from all four upland and peatland conifer stands. Gross ammonification rates were variable but similar in soils from uncut and cut peatland hollows (18–19mgNkg^–1day^–1) and upland forest floor soils (14–19mgNkg^–1day^–1). Gross ammonium ( ) immobilization rates were also variable but similar to ammonification rates. Median gross nitrification rates were within 0–2mgNkg^–1day^–1 in soils from all four upland and peatland cut and uncut stands, although rates were consistently higher for the soils from the cut stands. Large variability in gross nitrification rates were observed in peatland soils, however the highest gross nitrification rates were measured in saturated peatland soils. Net rates remained low in the soils from all four stands due to high nitrate ( ) immobilization and very fast turnover (<0.2 day). Our results suggest that potential losses may be negated by high immobilization in uncut and cut boreal forest stands. This study reveals the potential for the interaction of N production and consumption processes in regulating N retention in upland and peatland conifer forests, and the resilience of the boreal forest to disturbance.     

Effect of microrelief and vegetation on methane emission from wet polygonal tundra, Lena Delta, Northern Siberia

The effect of microrelief and vegetation on methane (CH₄) emission was investigated in a wet polygonal tundra of the Lena Delta, Northern Siberia (72.37N, 126.47E). Total and plant-mediated CH₄ fluxes were measured by closed-chamber techniques at two typical sites within a low-centred polygon. During the study period, total CH₄ flux averaged 28.0±5.4mgm^–2d^–1 in the depressed polygon centre and only 4.3±0.8mgm^–2d^–1 at the elevated polygon rim. This substantial small-scale spatial variability of CH₄ emission was caused by strong differences of hydrologic conditions within the microrelief of the polygon, which affected aeration status and organic matter content of the soils as well as the vegetation cover. Beside water table position, the vegetation cover was a major factor controlling CH₄ emission from polygonal tundra. It was shown that the dominant vascular plant of the study area, Carex aquatilis, possesses large aerenchyma, which serve as pathways for substantial plant-mediated CH₄ transport. The importance of plant-mediated CH₄ flux was strongly influenced by the position of the water table relative to the main root horizon. Plant-mediated CH₄ transport accounted for about two-thirds of the total flux in the wet polygon centre and for less than one-third of the total flux at the moist polygon rim. A clipping experiment and microscopic-anatomical studies suggested that plant-mediated CH₄ transport via C. aquatilis plants is driven only by diffusion and is limited by the high diffusion resistance of the dense root exodermes.     

Uptake of sewage derived nitrogen by<Emphasis Type="Italic"> Ulva rigida</Emphasis> (Chlorophyceae) in Bahía Nueva (Golfo Nuevo,Patagonia, Argentine)

Uptake kinetics of nitrogen derived from sewage–seawater mixtures (2.5–20% v/v effluent) were determined in the laboratory for Ulva rigida (Chlorophyceae) native from Bahía Nueva (Golfo Nuevo, Patagonia, Argentine). In terms of nitrogen concentration, experimental enrichment levels varied between 53.7 and 362.3M of ammonium and between 0.77 and 6.21M of nitrate+nitrite. Uptake rates were fitted to the Michaelis–Menten equation, with the following kinetic parameters: ammonium: V_max = 591.2molg^–1h^–1, K _s=262.3M, nitrate+nitrite: V _max=12.9molg^–1h^–1, K _s=3.5M). Both nutrients were taken up simultaneously, but ammonium incorporation was faster in all cases. The results show a high capability of Ulva rigida to remove sewage-derived nitrogen from culture media. In the field, most of the nitrogen provided by the effluent would be tied up in algal biomass, supporting low nitrogen levels found at a short distance away from the source.     

Change in soil carbon cycling for stand development of Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) plantations following clear-cutting

Soil carbon cycling was studied in Japanese cedar plantations with different stand ages after clear-cutting and was analyzed by a compartment model. The amount of biomass and the litterfall rate increased rapidly with the growth of Japanese cedar, which were approximated by a simple logistic function of stand age. The accumulation of A₀ layer decreased from 21tha^–1 to 5tha^–1 during the 10years following clear-cutting, and then recovered to nearly the same level as before clear-cutting within 20years after clear-cutting, although the amount of soil carbon in the mineral soil recovered more than 40years after clear-cutting. The total and mineral soil respiration rates increased rapidly after clear-cutting and gradually decreased in young stands and stabilized in old stands. The relative decomposition rate of the A₀ layer and organic matters in mineral soil was high in the young stands because of the relatively high soil temperature rather than the soil moisture content. After the closing up of the canopy, the relative decomposition rates of the A₀ layer and humus in the mineral soil stabilized at 0.14 to 0.16y^–1 and 0.005 to 0.013y^–1, respectively. Consequently, soil carbon cycling was strongly affected by clear-cutting. The amount of soil carbon rapidly decreased because of the cessation of litterfall and the increase of the relative decomposition rate of the A₀ layer and humus, and recovered gradually to the level before clear-cutting with the growth of the cedar plantation. The change in soil carbon cycling with stand development was partly caused by the change in soil temperature and moisture content but was mainly caused by the amount of cedar litterfall which changed significantly in the early stage of the stand following clear-cutting, and became slower and leveled off in the late stage with stabilization of the environmental conditions and litterfall rate.     

Transposome Insertional Mutagenesis and Direct Sequencing of Microbial Genomes

Preformed transposase-transposon complexes called Transposomes have been electroporated into bacterial cells. The magnesium dependent process of insertion of the transposable element into bacterial chromosomal DNA occurs in vivo. The transposition efficiency of a Transposome containing a kanamycin marker was between 1.0×10⁴and 1.0×10⁷kanamycin resistant clones per microgram of transposon DNA in three gram-negative enteric bacterial species. Transposon integration sites were examined by direct genome sequencing of chromosomal DNA. Genomic DNA was isolated from transposition clones and directly cycle sequenced with primers specific for the ends of the transposon. The precise location of genome interruption for a transposition clone was identified by homology to known genes or sequences. Mutant phenotypes were rapidly correlated with genomic insertions sites.     

Plant uptake of 14C-EDTA, 14C-Citrate, and 14C-Histidine from chelator-buffered and conventional hydroponic solutions

Chelator-buffered hydroponic solutions provide low and buffered free-metal concentrations and allow the easy calculation of nutrient species expected in these solutions. Some researchers suspect that the solutions allow plant uptake of chelates and that this uptake explains the failure of the free-ion activity model using these solutions. To determine the amount and method of chelate uptake, swiss chard was grown in solution culture in growth chambers for about three wks and then transferred to solutions containing ¹⁴C-EDTA, ¹⁴C-citrate, or ¹⁴C-L-histidine for a 21-hour assay. Much higher root and shoot ¹⁴C were found from treatments containing metabolites histidine (2706097 shoot Bq ¹⁴C) or citrate (2241953 shoot Bq ¹⁴C) than EDTA (280110 shoot Bq ¹⁴C). Passive transpirational flow could explain all of the EDTA uptake, but active uptake would be required to explain most of the citrate and histidine uptake even assuming some adsorption of ligand bound to roots. Swiss chard grown in solutions with the same total EDTA concentrations, but different amounts of Fe bound to EDTA, had 3-fold differences in root and shoot ¹⁴C concentrations. In a second experiment, swiss chard roots removed more EDTA from solutions containing mostly M-EDTA⁰ than M-EDTA^1- or M-EDTA^2- (288140, 245051, and 192559 Bq ¹⁴C, respectively) suggesting plant selectivity for EDTA and a non-apoplastic route of uptake or an effect resulting from root cell-wall adsorption. Results indicated buffering of metals by ligands allowed some ligand uptake with much more uptake occurring with metabolites citrate and histidine than EDTA. A passive or indiscriminate form of uptake does not appear to explain all EDTA uptake with a selectivity by swiss chard for M-EDTA complexes of lower charge.     

Phenotypic Differences in Buoyancy and Energetics of Lean and Siscowet Lake Charr in Lake Superior

At least two phenotypes of lake charr, Salvelinus namaycush, coexist in Lake Superior. A lean morph frequents the shallow inshore waters (< 50m) and the fat morph (siscowet) occupies the deeper offshore waters (50–250 m). The objective of this study was to determine if the elevated lipid concentration of siscowets reduces the costs of swimming in deep water. First, we modelled the effects of body composition (lipids) on the costs of swimming by lake charr, and then compared these theoretical results with empirical evidence obtained from Cesium 137-based estimates of food consumption, gross energy conversion, and swimming costs (activity multiplier). The attributes of growth, energy content (kJg^-1), lipid concentrations, and Cesium 137 concentration (Bqg^-1) were obtained from multimesh gillnet catches in eastern Lake Superior (1998 and 1999). The model showed that siscowet (fat) lake charr expended less energy than lean lake charr moving through the water column. Empirical evidence derived from Cesium 137 analysis confirmed that the activity multipliers of siscowets (fat) were less than those for lean charr. These findings support the view that the restoration of the fish community of the predominately deep water of the Great Lakes might be facilitated by the introduction of the fat phenotype.     

A Quantitative Analysis of the Courtship Acoustic Behaviour and Sound Patterning in Male Sand Goby, <Emphasis Type="Italic">Pomatoschistus minutus</Emphasis>

Many animal groups use sounds in reproduction in order to court mates or repel rivals. We describe the sounds and behavioural context of courtship sound production in male sand gobies, Pomatoschistus minutus, and examine the variability of acoustic parameters and the fine temporal patterning of sound units. Male sand gobies excavate a nest under a suitable solid substrate and attract females to mate, attaching the eggs to the ceiling of the nest. Before mating a female may repeatedly enter and leave a male's nest. Sounds were not detected during the courtship phase outside the nest, but were recorded when females were in the nest before spawning. Sounds were produced in 44–100% of such nest visits, varying with individual males. The sand goby sound consists of a train of pulses repeated at a rate of 23–29 pulses per second. The frequency spectrum of single pulses was continuous from 20–30Hz to 500Hz and reached a peak around 100Hz. The absolute sound pressure level ranged from 118 to 138dB re 1µPa at 1–3cm. The sand goby emits sound in distinct sound groups (bursts). Sound temporal features (duration, pulse repetition rate) vary systematically over the course of the burst. Within- and between-male variation of acoustic parameters was examined from sounds emitted by the male. Sound amplitude (peak-to-peak, mV) and pulse rate varied significantly among males, despite low individual stereotypy. Furthermore, sound pressure level correlated with body size. The potential informative content of acoustic parameters is discussed in the light of a possible role of the sand goby sound in mate choice.     

Estimation of molecular clocks for ITS and 28S rDNA in Erysiphales

Based on a comparative phylogenetic analysis of Goloviomyces and their host tribes of the Asteraceae, we speculate that Golovinomyces first acquired parasitism to the Asteraceae after migration of the family into the Northern Hemisphere and before the divergence of the tribe Carduaeae. The divergence time of the Carduaeae is estimated to be 25.2Myr ago based on the molecular clock of rbcL sequences of the Asteraceae. When 25.2Myr is given at the node of the first split of the phylogenetic tree of Golovinomyces, nucleotide substitution rates of the Erysiphales are calculated to be 2.52 × 10^–9 per site per year (0.01D = 3.97Myr) in the ITS region and 6.5 × 10^–10 per site per year (0.01D = 15.4Myr) in the D1 and D2 regions of the 28S rDNA.     

Sensory deficits induced by cadmium in banded kokopu, Galaxias fasciatus, juveniles

The potential for cadmium to produce sensory deficits in the mechanosensory lateral line and olfactory systems was examined in migratory Galaxias fasciatus juveniles or whitebait. Using a two-choice chamber apparatus, three groups of whitebait were tested for a known attraction to adult pheromones and then exposed to either 0.1, 0.5 or 1g Cd⁺² l^-1 for 48h and retested. The attraction to adult pheromones had been eliminated after exposure to concentrations of 0.5 and 1g Cd⁺² l^-1, indicating these levels of cadmium exposure had impaired olfactory function. Rheotaxis trials were conducted to determine the level of cadmium exposure which would inhibit lateral line function. The lateral line system was not blocked until a concentration of 2g Cd⁺² l^-1. The blocking of the lateral line and olfactory sensory systems was reversible. After 14 days recovery in clean freshwater both rheotaxis and the attraction to adult pheromones had returned. Whitebait were also tested for a preference/avoidance response at 2g Cd⁺² l^-1. Fish showed neither a preference for, or an avoidance of, a concentration which would disable both the lateral line and olfactory sensory systems. The disabling of these sensory systems may render migratory cues undetectable, affecting habitat selection by whitebait, which may ultimately affect the distribution of banded kokopu populations.     

Sucrose in storage media and cultivar affects post-storage regrowth of <Emphasis Type="Italic">in vitro</Emphasis> Hosta propagules

The goal of this research was to investigate if culturing in high sucrose (5%) liquid media during multiplication phase (stage II) would enhance endogenous sugar levels and dry matter sufficiently to allow storage of in vitro plants in sugar free media without adversely affecting post-storage recovery. Hosta tokudama Newberry Gold (NBG) and Hosta Striptease were cultured in Murashige and Skoog (MS) media containing 5% sucrose during stage II and transferred to rooting phase (stage III) in MS medium without (0%) sucrose or with 3% sucrose for 4weeks. At the end of stage III, cultures were stored, with the remaining media, at 10°C with 5molm^–2s^–1 photosynthetic photon flux (PPF) from cool white fluorescent lamps for 7 or 14weeks with or without a 2-week dark period prior to removal from storage. In both cultivars, stage III plants cultured in 3% sucrose media had higher soluble sugar levels and greater shoot and root biomass than those cultured in 0% sucrose media. Shoot and root soluble sugars decreased during storage. Shoot growth ceased during storage in both media. Root dry matter continued to increase in plants stored in 3% sucrose media but did not change in 0% sucrose media. Plants cultured in 3% sucrose media had less leaf chlorosis and less mortality after 7 or 14weeks of low temperature storage than the plantlets from sugar free media. Extending the storage period from 7 to 14weeks or introduction of 2-week dark period at the end of storage did not affect leaf chlorosis or plant mortality during acclimatization. Post-storage growth varied with the cultivar. Benefit of having sucrose in storage media was to develop a strong root system that aided the acclimatization and post-storage growth following 7 or 14week storage. Sucrose loading by culturing plants in liquid media containing 5% sucrose did not allow storage in sugar free media without adversely affecting post-storage growth in both cultivars.     

Protective effect of taurine on the detachment of cultured cardiac fibroblasts from the substratum induced by calcium depletion

Summary. Removal of Ca²⁺ from the incubation medium of cultured rat cardiac fibroblasts causes cellular morphological changes, such as the formation of blebs, the ballooning of the cell membrane and the detachment from the culture dish. A 24hr preincubation with 20mM taurine blocked the Ca²⁺ depletion-induced detachment of the cardiac fibroblasts. However, taurine treatment did not prevent other morphological changes induced by Ca²⁺ depletion. The data suggest that taurine plays an important role in cell adhesion in the heart.