Optimal assay conditions for aspartate transcarbamylase (ATCase) activity in mesozooplankton

The optimal conditions for storing and assaying mesozooplanktonaspartate transcarb amylase (ATCase) (EC 2.1.3.1[EC].) are definedin order to enable the specific activity of this enzyme to beused as an index of mesozooplankton productivity. ATCase activitywas found to be stable for more than 2 weeks when stored inliquid nitrogen, with a negligible loss of activity, whateverthe state of frozen material (cell-free homogenate or wholecells). As an alternative, –20C, –90C freezersand dry ice (–80C) can give more flexibility for storingand transporting material, provided initial freezing has beencarried out at –196C. ATCase specific activity was stableover a large range of total protein concentrations (2–55mg ml^–1) for most mesozooplankton samples, and the enzymeactivity was linear between 0.8 and 80 nmol carbamyl aspartatemin^–1 mg^–1 of total protein. The optimal incubationtime (30 min), temperature (35C) and pH range (9–9.5)for assay of ATCase were similar forCalanus helgolondicus (Crustacea:Copepoda) (G.O.Sars) and three mesozooplankton assemblages sampledoff Plymouth (UK) in May, August and November. However, theKm values for both substrateswere variable: from 3.9 to 79.1for aspartate, and from 0.83 to 3.46 for carbamyl phosphate.Such variations arelikely to be due to the presence of differentATCase catalytic potential, rather than changes in ATCase assemblage.     

Temporal variability in copepod fecundity during two different spring bloom periods in coastal waters off Plymouth (SW England)

The temporal variation in egg production of the planktonic copepodsCalanus helgolandicus, Temora longicornis and Pseudocalanuselongatus was studied during two different spring bloom periodsin 1989 and 1990 by weekly sampling at two permanent stationsin coastal waters off Plymouth (SW England). Copepod egg productionwas estimated in situ by incubating individual adult femalesin filtered seawater for 24 h (72 h until hatching for P.elongatus)at the field surface temperature. The relationship between copepodegg production rates and: (i) chlorophyll a concentration (totaland >10 µm size fraction), (ii) temperature and (iii)initial copepod gut pigment content was investigated. The springbloom periods were very different in both years, with the occurrenceof a Phaeocystis sp. bloom in 1990, which negatively affectedthe feeding and fecundity of copepods. Egg production ratesin spring 1989 were significantly correlated with chlorophylla concentration (particularly with the >10 µm fraction),field temperature and copepod gut pigment contents. In spring1990, egg production rates were also correlated with copepodgut pigment contents, but no significant correlations were obtainedwith temperature or with chlorophyll a concentration, as a consequenceof the lower egg production rates obtained during the Phaeocystissp. dominance period. These results show that food availabilityis the factor which mainly affects the fecundity of neriticcopepods in short time periods.     

Grazing impact of micro- and mesozooplankton during a spring situation in coastal waters off the Gironde estuary

The aim of this study was to characterize the grazing link betweenphytoplankton and zooplankton. Quantifying rates of phytoplanktonutilization by micro- and mesozooplankton is an attempt to understandthe control of phytoplankton populations in marine ecosystemsand the export flux of organic matter to the depths. Sampleswere taken in the plume of dilution of the Gironde estuary duringspring (May 1995). The mesozooplankton community was dominatedby the heterotrophic dinoflagellate Noctiluca scintillans andby copepods. Our estimates showed that on average, 26% of thetotal primary production was grazed daily by mesozooplankton.However, the pressure exerted by microzooplankton on the primaryproduction was estimated to be approximately four times higher.As a consequence, during this spring situation in the plumeof dilution of the Gironde estuary when small cells dominated,microzooplankton represented the most important grazing communityand could potentially control the growth of phytoplankton.     

Estimations of mesozooplankton biomass in a coastal upwelling area off NW Spain

The abundance and several estimators of mesozooplankton biomass(dry weight, ash-free dry weight, displacement volume, carbonand nitrogen) were measured in a coastal upwelling area offLa Coru*****na (NW Spain) at monthly intervals between 1990and 1995. Holoplanktonic copepods dominated in most of the samples,but meroplanktonwere also important near the coast, especiallyduring late spring and through summer. Gelatinous organisms(medusae, siphonophores and larvaceans) reached significantabundances (-10%) in shelf waters by late summer, but also nearthe coast in spring. Log-linear equations were computed betweendry weight, displacement volumeand the other biomass estimators,to allow for interconversion among different measurements. Toprevent the influence of meroplankton and gelatinous zooplanktonin the estimations of biomass, the equations were adjusted usingonly samples in which the abundance of each of these groupswas <10% of total individuals. Our equations produced carbonand ash-free dry weight estimates that are comparable to thoseobtained with the ratiosgiven in other studies in the region.However, the esti mations of mesozooplankton carbon from displacementvolume using the adjusted equation with our data are lower thanthose reported previously for the area, which werecomputed usingequations adjusted with data from a different ecosystem. Theuse of interconversion factors taken from the literature isdiscussed, along with the implications of the estimations ofmesozooplankton biomass in the computations of the flux of carbonthrough the ecosystem.     

Egg production of individual copepods of Acartia grani Sars from coastal waters: seasonal and diel variability

Seasonal and diel fecundity patterns of adult Acartia granifemales from a coastal area of Malaga Bay (south of Spain) werestudied between spring and autumn 1990. Copepod egg productionwas measured in situ by short-term (     

Control of aspartate transcarbamylase activity in type 5 adenovirus-infected HeLa cells

Consigli, Richard A. (University of Pennsylvania, Philadelphia), and Harold S. Ginsberg. Control of aspartate transcarbamylase activity in type 5 adenovirus-infected HeLa cells. J. Bacteriol. 87:1027-1033. 1964.-Type 5 adenovirus infection induces increased aspartate transcarbamylase (ATCase) activity during the period of magnified nucleic acid biosynthesis. Increased activity can be prevented by addition of pyrimidines to the culture medium. ATCase in HeLa cells is regulated by feedback inhibition, and purified enzyme can be inhibited in vitro by cytidine triphosphate (CTP). The enzyme from infected cells has a pH optimum, maximal velocity, and K(m) for aspartate distinctly different from ATCase from control cells. However, heating of ATCase from uninfected cells converts the enzyme so that its characteristics are identical with enzyme from infected cells. Conversely, addition of CTP to ATCase from infected cells changes the characteristics of the enzyme so that they are the same as those of enzyme from uninfected cells. The evidence presented suggests that increased nucleic acid biosynthesis in infected cells initiates a release from feedback inhibition and increases ATCase activity by reducing the concentration of pyrimidines and purines in the acid-soluble pool.     

Phytoplankton biomass and production in shelf waters off NW Spain: spatial and seasonal variability in relation to upwelling

Summary Chlorophyll-a and primary production on the euphotic zone of the N-NW Spanish shelf were studied at 125 stations between 1984 and 1992. Three geographic areas (Cantabrian Sea, Rías Altas and Was Baixas), three bathymetric ranges (20 to 60 m, 60 to 150 m and stations deeper than 200 m), and four oceanographic stages (spring and autumn blooms, summer upwelling, summer stratification and winter mixing) were considered. One of the major sources of variability of chlorophyll and production data was season. Bloom and summer upwelling stages have equivalent mean and maximum values. Average chlorophyll-a concentrations approximately doubled in every step of the increasing productivity sequence: winter mixing — summer stratification — high productivity (upwelling and bloom) stages. Average primary production rates increased only 60% in the described sequence. Mean (± sd) values of chlorophyll-a and primary production rates during the high productivity stages were 59.7 ± 39.5 mg Chl-a m^–2 and 86.9 ± 44.0 mg C m^–2 h^–1, respectively. Significant differences in both chlorophyll and primary production resulted between geographic areas in most stages. Only 27 stations showed the effects of the summer upwelling that affected coastal areas in the Cantabrian Sea and Rías Baixas shelf, but also shelf-break stations in the Rías Altas area. The Rías Baixas area had lower chlorophyll than both the Rías Altas and the Cantabrian Sea areas during spring and autumn blooms, but higher during summer upwelling events. On the contrary, primary production rates were higher in the Rías Baixas area during blooms in spring and autumn. Mid-shelf areas showed the highest chlorophyll concentrations during high productivity stages, probably due to the existence of frontal zones in all geographic areas considered. The estimated phytoplankton growth rates were comparable to those of other coastal upwelling systems, with average values lower than the maximum potential growth rates. Doubling rates for upwelling and stratification stages in the northern and Rías Altas shelf areas were equivalent, despite larger biomass accumulations during upwelling events. Low turnover rates of the existing biomass in the Rías Baixas shelf in upwelling stages suggests that the accumulation of phytoplankton was due mainly to the export from the highly productive rías, while the contribution of in situ production to these accumulations was relatively lower.     

Characterization of aspartate transcarbamylase activity from gonads of the soft shell clam,Mya arenaria

Aspartate transcarbamylase (ATCase, EC 2.1.3.2) has been shown to be a good index of the reproductive cycle in marine molluscs. However, this enzyme has never been studied in the soft shell clam Mya arenaria. The characteristics of gonadal ATCase of the soft shell clam, Mya arenaria were therefore determined since we need powerful tools to assess the degree of effects of endocrine disruptors in this species at risk. Enzyme kinetic values observed at pH 8.3 were significantly lower than those measured at pH 9.4. The optimal conditions for the enzyme assays were reached in the presence of a 10 mM of substrate concentration and at pH 9.2 for 60 min at 37 °C. We have found that the enzyme was heat sensitive, markedly activated by DMSO and DMF, but no effect was observed with ethanol, ATP or CTP. However, clam ATCase activity was partly inhibited by the addition of CuSO₄ and PHMB to the medium, an inhibition that could be attributed to the presence of SH sites in cysteine residues localized in the catalytic site of this enzyme. All these results will be very useful in the near future to study the gametogenetic process of Mya arenaria, since little is known about the factors that control the physiological process of reproduction in this bivalve of ecological and economic importance. Studies of variations of the activity of aspartate transcarbamylase will also be useful as a potential biomarker to evaluate the disruption of gametogenesis in clams exposed to endocrine disruptors in situ.     

Biological monitoring of heavy-metal contamination in coastal waters off Casey Station, Windmill Islands, East Antarctica

Heavy-metal concentrations were determined in tissues of different species of benthic invertebrates collected in the Casey region (Australian Antarctic Territory) where an old waste-disposal tip site is a source of contamination. The species studied included the bivalve Laternula elliptica, starfish Notasterias armata, heart urchins Abatus nimrodi and A. ingens and gammaridean amphipod Paramoera walkeri. The specimens were collected at both reference and contaminated locations where lead was the priority element and copper was the next most important in terms of increased concentrations. The strong association between a gradient of contamination and concentrations in all species tested indicates that they are reflecting well the environmental changes, and that they appear as appropriate biological indicators of heavy-metal contamination. Aspects of the biology of species with different functional roles in the marine ecosystem are discussed in relation to their suitability for wider use in Antarctic monitoring programmes. For example, in terms of heavy-metal bioaccumulation, the bivalve appears as the most sensitive species to detect contamination; the starfish provides information on the transfer of metals through the food web while the heart urchin and gammarid give indications of the spatial and temporal patterns of the environmental contamination. The information gathered about processes of contaminant uptake and partitioning among different tissues and species could be used in later studies to investigate the behaviour and the source of contaminants.     

Spatio-temporal structure of tidally mixed coastal waters: variability and heterogeneity

In the tidally mixed coastal waters of the eastern English Channel,the vertical properties of temperature, salinity, in vivo fluorescence,turbidity and light were investigated along an inshore-offshoretransect, sampled in different tidal conditions. The verticaldistribution of these parameters was then characterized in termsof mean value, variability (i.e. standard deviation) and heterogeneity(i.e. fractal dimension), regarded as a quantitative characterizationof the structure of the vertical variability of both physicaland biological parameters. These different estimates were thensubjected to multivariate spatio-temporal analysis and showeda very specific spatio-temporal structure suggesting a differentialcontrol of the vertical properties of water masses associatedwith both inshore-offshore gradient and tidal advection. Inparticular, the fractal dimension (i.e. heterogeneity) of invivo fluorescence is higher during ebb for offshore waters,suggesting a density dependence of the phytoplankton biomassstructure. In contrast, the vertical variability of fluorescenceis higher during flood for inshore waters, leading to an inverserelationship between variability and heterogeneity of the verticaldistribution of phytoplankton biomass.     

A kinetic model of cooperativity in aspartate transcarbamylase.

A relatively simple kinetic model is proposed to account simultaneously for data on the binding of carbamyl phosphate and succinate to aspartate trans carbamylase (ATCase), and for the relaxation spectrum associated with this binding. The model also accounts for measurements of the initial velocity of the reaction of ATCase with respect to aspartate and carbamyl phosphate. The principal assumption made is that ATCase consists of three identical noninteracting cooperative dimers. Ordered binding and both sequential and concerted conformational changes in the dimers are needed to account for the properties of ATCase. The values of the parameters of this model can be determined by fitting to existing experimental evidence. Various new quantitative predictions are made that can serve as additional tests of the proposed theory.     

In situ activity of NAC11-7 roseobacters in coastal waters off the Chesapeake Bay based on ftsZ expression

Determining in situ growth rates for specific bacterioplankton is of critical importance to understanding their contributions to energy and matter flow in the Ocean. Quantifying expression of genes central to cell division is a plausible approach for obtaining these measurements. In order to test this approach's assumptions, a quantitative PCR assay targeting the cell division gene ftsZ in the ubiquitous NAC11-7 group of the Rhodobacterales order of marine bacteria was developed. ftsZ genes and their corresponding mRNAs were measured in diel in situ samples and in parallel on-deck incubations. Strong correlations between ftsZ expression and gene abundance (R-squared = 0.62) were observed in situ. Rapid changes in NAC11-7 ftsZ gene copies suggested that different populations from different water types were sampled with a significant positive correlation between ftsZ expression and water temperature (R-squared = 0.68, P < 0.001). An outlier to this trend occurred at a single time point (9:00), which was remarkably consistent with a concomitant peak in ftsZ expression in on-deck incubations, suggesting the possibility of synchronous population growth.     

Influence of UV-B radiation on bacterial activity in coastal waters

The impact of UV-B radiation (290–315 nm) on bacterialactivity and abundance in coastal water was studied in mesocosmexperiments in May 1994 and May 1995 at Kristineberg MarineResearch Station, Sweden. Mesocosms (6 m³) containing naturalpelagic communities were exposed either to ambient irradiation(AMB), ambient irradiation with enhanced UV-B (+UV) (0.7 W m^–24 h every day around noon), or ambient irradiation screenedfor UV-B (–UV). Bacterial activity in the mesocosms wasmeasured by means of thymidine incorporation in short-term testsduring incubations at ambient irradiation, at ambient irradiationwith enhanced UV-B, and at ambient irradiation screened forUV-B. In +UV mesocosms, bacterial activity was significantlystimulated when incubated at ambient radiation. The stimulatingeffect was suggested to be due to an increase in carbon or nutrientsupply through a photodegradation of recalcitrant dissolvedorganic material (DOM). Low attenuation coefficients for UV-Band PAR (400–700 nm) in the +UV mesocosms supported thishypothesis. The bacterial activity in +UV mesocosms, however,was inhibited when incubations were made at enhanced UV-B irradiation,implying that the bacteria had become more sensitive to UV-Bradiation. The increased sensitivity to UV-B exposure in bacterialassemblages that already had been exposed and stressed by UV-Bradiation is suggested to be due to an overburdening of theenergy-consuming DNA repair mechanism. The data suggest thatincreased UV-B radiation, which might occur with ozone depletion,may both stimulate and suppress bacterial activity in coastalwaters, implying that the net outcome of enhanced UV-B radiationcould be an unchanged bacterial activity.     

Use of environmental DNA in early detection of Mnemiopsis leidyi in UK coastal waters

Biological Invasions - Since 2005, the ctenophore Mnemiopsis leidyi has been reported as present in northern Europe. Although the impacts of this voracious predator on North Sea fisheries remain...