Measurements of electron transport activities in marine phytoplankton

A simple and sensitive spectrophotometric method of measuring electron transfer in the transport system (ETS) in marine phytoplankton has been developed and characterized. The assay is based on the reduction of the tetrazolium salt 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) by homogenates in the presence of the nonionic detergent Triton X-100. The simplicity and sensitivity of this assay have considerable advantage over existing methods of measuring ETS activity in marine samples where numerous assays must be rapidly carried out on samples which often demonstrate low activities. Several established substrates and inhibitors of electron transport in the ETS were investigated, and each is discussed in relation to the site of INT reduction in the diatoms Skeletonema costatum and Chaetoceros debilis and in the unicellular green alga Dunaliella tertiolecta. The results from these studies suggest that the maximum rate of electron transport, V _max, is measured in each case.     

The production and secreton of digestive enzymes in the purple seastar Pisaster ochraceus

The purple seastar Pisaster ochraceus contains clearly measurable protease and amylase activity. Centrifuged supernatants of pyloric caeca homogenates undergo a spontaneous threefold increase in protease activity when incubated under toluene for 50 h at 25°C. Amylase activity remains nearly constant over this period. Bovine trypsin at a 1 to 100 ratio (trypsin to supernatant protein) induces a twofold increase in protease activity over that of the control supernatant while having virtually no effect on amylase activity over the control. The data indicate a specific interaction of trypsin with a protease zymogen rather than a conspecific hydrolysis of membrane components or vesicles by trypsin. Two percent Triton X-100 used as a diluent in place of distilled, deionized water causes a sevenfold increase in protease activity and a twofold increase in amylase activity in pyloric caeca supernatants. The use of Triton as a diluent in the preparation of a stomach-tissue supernatant allows quantitative measurement of both protease and amylase activity in that tissue.     

Respiration and physiological state in marine bacteria

The relationship between oxygen consumption (R) and respiratory electron-transport-system (ETS) activity was investigated in batch cultures of 5 species of marine bacteria, Vibrio adaptatus, V. anguillarum, a partially identified Vibrio sp. SA774, Serratia marinorubra, and Pseudomonas perfectomarinus. Although cellular levels of R and ETS varied widely among the species tested, the R:ETS ratios for growing or senescent populations were relatively constant among the species; these ratios were 5.02 in growth and 0.426 in senescence, with coefficients of variation of 29 and 20%, respectively. The lower senescent-phase R:ETS ratio was due to a depression of the respiration rates following growth termination. The regression log (R per cell) = 0.832 log (ETS per cell) + 0.727 for the growing populations was similar to that determined for marine zooplankton. The slight dependency of the R:ETS ratio on organism dry weight found for zooplankton was supported by our data. Planktonic respiration rates estimated from measured ETS-depth profiles in the eastern tropical North Pacific Ocean using the senescent-phase R:ETS ratio were similar to published oxygen consumption rates in the deep sea.Contribution No. M79-61 from the University of Washington's Department of Oceanography, and No. 79032 from the Bigelow Laboratory for Ocean Sciences.     

Dehydrogenases as tools in the study of marine sediments

A method is outlined for the measurement of NADH₂-dependent dehydrogenase activity in whole samples as well as in fractions of carbonate sediments. The method is based on extraction of enzyme activity with phosphate buffer containing 0.12% Triton X-100, dialysis, and a photometric assay which permits initial velocity determinations of the enzyme reaction under controlled conditions of pH, temperature, and substrate concentration. The dehydrogenases extracted from carbonate sediment of a subtropical beach at Bermuda are characterized by low K _m values (0.03 to 0.07 mM NADH₂/1), pH maxima around 8.5, and temperature maxima between 35° and 40°C. Their vertical distribution in the sediment agrees with what is known about the distribution of biomass on beaches of this type, maximum activity occurring in the topmost centimetre. Approximately 80 to 90% of the total electron transport activity is contained in the grain fraction, the remainder in the interstitial fraction of a sediment sample. From this it can be inferred that the interstitial fauna contributes only little to the total energy budget of this type of sediment, which is dominated by the Aufwuchs on the sand grains. The electron transport activity (ETA) measured by means of the dehydrogenase assay is taken to represent the maximum transport capacity (ETC) of all the organisms living in the sand ecosystem. From published evidence it is inferred that oxygen consumption of sediment samples and of individual organisms measured in vivo usually accounts for 10 to 20% of the maximum electron transport capacity measured in vitro with saturating concentrations of substrates and with Triton X-100 as part of the enzyme assay. It is suggested further that the ratios of dehydrogenase activities in different fractions or zones of sediment, or of different organisms, may represent one of the best indicators, so far available, of energy relationships in marine sediments.Contribution No. 652 from the Bermuda Biological Station for Research.     

Zooplankton biomass and indices of grazing and metabolism during a late winter bloom in subtropical waters

The development of the so-called late winter bloom in subtropical water was studied in an oceanic area north of the Canary Islands from January to May 2000. Zooplankton was sampled at short-term intervals (1–4 days) during the bloom (January–March), and biomass, indices of grazing (gut fluorescence) and metabolism (electron transfer system activity, ETS) were measured in four different size fractions (100–200, 200–500, 500–1000 and >1000 µm). During the bloom, ETS activity and gut fluorescence increased before the development of zooplankton biomass. At the end of February, the presence of an impressive cloud of dust formed in the Sahara desert was related to an increase in chlorophyll and small zooplankton a week later. The increments in biomass were the consequence of consumption by zooplankton as inferred from the indices of grazing and metabolism. Estimated grazing from gut fluorescence and gut evacuation rates during the period of study accounted for 55% of the assessed total ingestion from respiration and normal values of assimilation, showing the importance of the non-pigmented food in the diet of zooplankton in these waters. In contrast, the sharp decreases in zooplankton biomass observed during the bloom appeared during the dark period of the moon, the days in which the diel vertical migrants reach the shallower layers, in agreement with previous works in the area. Thus, the development of the late winter bloom in this region is suggested to be driven by the interplay between resource and consumer controls.Communicated by S.A. Poulet, Roscoff     

非离子表面活性剂淋洗萘污染土壤的修复研究

采用土柱实验研究了非离子表面活性剂对焦化厂萘污染砂土的淋洗效果,同时研究了超声预处理对非离子表面活性剂土壤淋洗的增强效果及粉土和粘土对非离子表面活性剂土壤淋洗的不利影响.研究表明,1)1g·L-1的TritonX-100、AEO-9和Tween80对同一萘浓度(192.4mg·kg-1)的土壤洗脱效率分别为73.0%、81.5%和59.0%,2g·L-1的表面活性剂的洗脱效率要高于1g·L-1的洗脱率,分别为96.5%、95.1%和88.2%.2)对土壤进行短时超声处理(80Hz)促进了洗脱率的提高,超声10min的淋洗效果要高于超声5min的淋洗效果;土壤分别经5min和10min超声处理后,2g·L-1浓度的TritonX-100对萘的洗脱率从94%分别提高到98.8%和99.6%,洗脱时间从255min分别减少到172min和160min.3)砂土中含有粉土和粘土对洗脱效果有不利影响,且粘土的影响要更严重,当萘污染的砂土混入5%的粘土时,2g·L-1浓度的TritonX-100对萘的洗脱率从94.7%下降至73.8%,洗脱时间从255min增加至605min,而混入20%的粉土洗脱率仅下降到88.4%,洗脱时间增加至512min.     

The spring phytoplankton bloom in Lindåspollene,a land-locked Norwegian fjord. II. Biomass and activity of net- and nanoplankton

From February 24 to April 24, weekly samples were collected at fixed depths at one station in Lindåspollene, a land-locked Norwegian fjord. Adenosine triphosphate (ATP), chlorophyll a, phaeophytin, ¹⁴C assimilation, and respiratory activity [electron transport system (ETS) activity] were measured in the net- (>30 m) and nanoplankton. Netplankton contained on the average 48% of the total chlorophyll a and 56% of the ATP, but contributed only 7% to the total carbon assimilation and 11% to the ETS activity. The assimilation numbers for net- and nanoplankton ranged from 0 to 1.2 and from 1.5 to 13.2, respectively. At the oxygen/hydrogen sulphide interface, high concentrations of ATP, but not of chlorophyll a, were found in the nanoplankton fraction. Netplankton algae grew actively only in the first phase of the bloom, and nanoplankton predominated later, apparently due to low nutrient concentrations. During the bloom, Skeletonema costatum made up the main part of the biomass. The number of cells in the chains decreased throughout the bloom, possibly reflecting the lowered silicate content. It appeared that only nanoplankton were grazed by zooplankton, while netplankton sank to the bottom and represented input to the benthos.     

Effects of food-level acclimation on digestive enzyme activities and feeding behavior of Calanus pacificus

A recent hypothesis in the zooplankton literature states that zooplankton acclimate to ambient food concentrations such that higher digestive enzyme activities and, consequently, higher maximum ingestion rates are achieved at higher food levels. To test this hypothesis, adult female Calanus pacificus, collected from the main basin of Puget Sound, Washington, USA, in August 1979 and May 1982, were conditioned for 2 wk at different concentrations of the diatom Thalassiosira weissflogii (=fluviatilis). Ingestion rates and the activity of the digestive enzymes laminarinase, maltase, and cellobiase were measured periodically during acclimation and in a block-designed feeding experiment at the end of acclimation. Consistent with the hypothesis, maximum ingestion rate and digestive enzyme activity were positively correlated. However, in contrast to the hypothesized mechanism, this result arose because both maximum ingestion rate and digestive enzyme activity were negatively correlated with food concentration during acclimation. The enhanced ingestion of copepods following long-term (12 to 14 d) acclimation to low food is similar to that previously described for short-term (e.g. 1 d) starvation. It might be energetically optimal for copepods experiencing a patchy food environment to maintain higher levels of digestive enzymes at low food concentrations in order to exploit high concentrations of food when encountered.     

Description and use of an improved method for determining estuarine zooplankton grazing rates on phytoplankton

A method of rapidly determining zooplankton grazing rates on natural mixed phytoplankton populations using ¹⁴C is described. The method simplifies the design of grazing experiments as the grazing time can be kept short enough to prevent recycling of the isotope, and growth of the phytoplankton substrate. Very high specific activity, ¹⁴C-labelled phytoplankton concentrated either by centrifugation or sieving, may be used either as the sole grazing substrate, or as a tracer in natural mixed phytoplankton. Zooplankton, confined in glass jars at either ambient, or higher than ambient concentrations, are permitted to feed on the phytoplankton for periods of 30 min and 2 h, and are then separated by sieving. The zooplankton community grazing rate, or, if the samples are sorted into species, the individual species grazing rates, can be determined after scintillation counting of the zooplankton. The rate of appearance of ¹⁴C-labelled phytoplankton in the zooplankton is an estimate of the grazing rate, and the slope of the line joining the grazing rates at various phytoplankton concentrations gives an estimate of the grazing rate constant for the zooplankton population. The method provides a quick way of obtaining both zooplankton population, and individual species grazing rates on natural mixed phytoplankton. In two experiments, labelled phytoplankton was used as the sole grazing substrate in concentrations ranging between 0.4 and 5 times ambient levels. Grazing rate constants, for net-caught zooplankton concentrated to 46 times (Experiment 1) and 28 times (Experiment, 2) ambient estuarine levels were-0.14and-0.12 of the phytoplankton standing stock per day, respectively. There was a linear increase in the amount of phytoplankton grazed with an increase in phytoplankton concentration up to four times ambient phytoplankton levels. When tracer amounts of labelled phytoplankton were added to samples containing both phytoplankton and zooplankton at ambient concentrations the grazing rate constants were-0.28 and-0.42 of the phytoplankton standing stock per day. We conclude that zooplankton grazing was the major control factor of phytoplankton population size during October–November 1975 in South West Arm, Port Hacking, near Sydney, Australia.     

A fast and reliable method for quantitative determination of total mercury in vegetables

A cloud-point extraction (CPE) process using the nonionic surfactant, polyethylene glycol tert octylphenyl ether (Triton X-114) was employed for determination of Hg(II) ions in aqueous solutions. The method is based on the ion-pairing reaction of Hg(II) with Pyronin B (PyrB⁺) in the presence of excess iodide at pH 6.0 and extraction of the complex formed. The chemical variables affecting CPE efficiency were studied, and the analytical characteristics of the method were obtained. The calibration curves were linear in the range of 1–40 μg L^?1 with the detection limits of 0.35 and 0.30 μg L^?1 at 556 and 521 nm. Selectivity was also tested. The coefficients of variation of the method are 2.4% and 5.2% for five replicate measurements of mercury at levels of 10 and 25 μg L^?1, respectively. The results obtained for two certified reference samples were in a good agreement with the certified values. The method was applied to the determination of total mercury in vegetable samples.     

Are closure terms appropriate or necessary descriptors of zooplankton loss in nutrient–phytoplankton–zooplankton type models?

Our current knowledge of plankton ecology ascribes a large proportion of zooplankton losses to zooplankton cannibalism and carnivory, rather than via the activity of higher trophic levels beyond the plankton. However, planktonic ecosystem models, such as the widely used nutrient–phytoplankton–zooplankton (NPZ) type models, typically represent all zooplankton losses by mathematically (rather than biologically) justified closure functions. Even where it is assumed that these closure functions include zooplanktonic cannibalism and carnivory, these processes are not explicitly implemented within the grazing function of the zooplankton. Here it is argued that this representation of zooplankton losses through “closure” terms within planktonic food web models is neither appropriate nor necessary. The general consequences of implementing a simple function incorporating zooplankton cannibalism and carnivory (intra-guild predation) within a planktonic food web model are compared against models implementing different types of traditional closure functions. While the modelled biomass outputs may appear similar, the fate of annual primary production and f-ratios vary widely. There appears no justification for the continued use of traditional closure term to depict zooplankton loss processes on biological or modelling arguments. To do so can seriously misrepresent the fate of primary production and thence trophic dynamics.     

Calibration for the determination of 19 trace elements in serum and urine

A fast, high-throughput and accurate method was developed for determination of Al, As, Ba, Cd, Co, Cr, Cs, Cu, Fe, Mn, Ni, Pb, Se, U, Zn, Sb, Sn, I and Hg in urine and serum by inductively coupled plasma mass spectrometry. The samples were directly analyzed after 1/20 (v/v) dilution in 0.4% (v/v) HNO₃ and 0.005% Triton X-100. Three calibration modes were tested: aqueous and matrix matching with urine and serum. The accuracy was tested using reference materials of serum, urine and spiking. Results showed that the use of matrix matching calibration reduced the interferences and improved the recoveries for Al, Co, Pb and I in urine. The matrix matching did not affect the results considerably for serum. When serum was spiked with As, Co, Cs, Pb, U, Hg, I, Ba, Al, Cr and Ni, only matrix matching presented good recoveries. Helium was used as a collision cell gas reducing effectively polyatomic interferences for Al, Mn, Cu, Ni, Cr, Zn, As and Fe. Selection of the best internal standard was carried out for each element. The use of diluted HNO₃ improved the limit of detection. Finally, the method was applied successfully in samples of urine from workers occupationally exposed.     

A novel ligand for cloud point extraction to determine gold content in ore samples

Gold is a valuable metal occurring usually at very low concentrations in complex natural samples. Gold analysis thus needs preconcentration methods. Classical liquid–liquid extraction involves hazardous organic solvents. Alternatively, cloud point extraction involves non-toxic and nonvolatile surfactants. Here, we analyzed ore samples. The complex of Au(III) with sulphapyridylazo resorcin was extracted by the cloud point method. Concentrations were measured by flame atomic absorption spectrometry. We tested the effects of pH, reagent concentration, Triton X-100 surfactant concentration, equilibration temperature and time. Results show that the best detection limit of the method was 0.48 µg L^?1 for Au with a preconcentration factor of 31 times. Calibration is linear in range of 6.4–2,000 µg L^?1, and relative standard deviations are lower than 5 %. Quantum chemical computations reveal the plausible structure of the gold–ligand complex. This report represents the first determination of gold using sulphapyridylazo resorcin as a ligand.     

Parameterising competing zooplankton for survival in plankton functional type models

Marine plankton ecosystems are an important component of biogeochemical cycling in the oceans. Operational plankton functional type (PFT) models, that group plankton according to their biogeochemical properties, are currently being developed to resolve biogenic gas exchange between the ocean and atmosphere, and to model the lowest trophic levels in fisheries models. A fundamental problem with these models is that PFTs often go extinct in computer simulations, effectively removing the biogeochemical processes from the models. Cropp and Norbury [Cropp, R., Norbury, J., 2009a. Parameterizing plankton functional type models: insights from a dynamical systems perspective. J. Plankton Res. 31, 939-963] demonstrated that parameter combinations that allowed all PFTs to stay extant for all time in stable, homogeneous environments were rare in a PFT model with two competing phytoplankton and one zooplankton (NP₁P₂Z). In this paper, we examine the dynamical properties of a generic predator-predator-prey PFT model, and apply the analysis techniques developed by Cropp and Norbury to a simple example PFT model with one phytoplankton and two zooplankton (NPZ₁Z₂) in order to explore its properties and parameter space. We find that the properties of predator-predator-prey PFT systems are fundamentally different from those of predator-prey-prey PFT systems. The likelihood of parameter combinations for which all PFTs stay extant for all time in predator-predator-prey PFT systems depends critically on the process formulations used, and the properties of co-existing zooplankton (as defined by their parameter values) are quite different to those of co-existing phytoplankton.     

The use of measurable coefficients in process formulations—zooplankton grazing

This paper redefines a construct previously used to model phytoplankton—zooplankton interactions in such a way as to permit the use of measurable quantities as contruct coefficients. The new construct can use unaltered values of the half-saturation constant for zooplankton grazing on total available food (k_s) and the minimum food concentration necessary to stimulate effective feeding (BMIN) reported in the literature. Typical values for these coefficients are 0.1–15 and 0.016–0.19, respectively.     

A plankton study in the eastern Mediterranean Sea

A synoptic study of the phytoplankton and zooplankton distribution in the eastern Mediterranean Sea was carried out in the summer of 1965 during a cruise of R.V. “Pillsbury”. The phytoplankton maximum was normally found at 100 m, below the 1% level of surface light, with a frequent smaller peak at 40 m. In shallower waters, the main peak was at 80 m. The greatest number of phytophagous crustaceans was recorded at, or close to, the phytoplankton maximum. Depth relationships of phytoplankton and functionally phytoplanktonic species are discussed. Zooplankton was collected from depths down to 4,400 m, the deepest cast being in the Rhodes Deep. The most common species and genera of some selected groups of zooplankton were identified and their distribution considered in the light of some previous cruises in part of the area. The presence of Acantharia containing zooxanthellae with chlorophyll at, and below, 4,000 m was recorded for the first time. Potentially photosynthetic Ceratium vultur and C. carriense (Dinophyceae), both species in active division, were also found at these depths, as well as Halosphaera viridis (Prasinophyceae), which was recorded down to a depth of 1,000 m.     

Calculation and use of selectivity coefficients of feeding: Zooplankton grazing

A straightforward method for calculating selectivity coefficients (W_ij) of predation from raw data, mortality rates of prey, filtering rates, feeding rates and electivity indices is derived. Results from a comparison of selectivity coefficients for the copepod Diaptomus oregonensis grazing under a number of experimental conditions suggest that W_ij's for size-selective feeding are invariant, a conclusion also supported by the leaky-sieve model. Recommendations are made on how to use W_ij's in linear and nonlinear feeding constructs for zooplankton and other animals.     

Concentration of ascorbic acid and antioxidant response in early life stages of Engraulis ringens and zooplankton during the spawning seasons of 2006–2009 off central Chile

This study reports changes in ascorbic acid (AA) in anchoveta eggs, copepods and zooplankton during the 2006, 2007 and 2009 main spawning seasons in the coastal area of the central Humboldt Current System, Chile. Anchoveta eggs, copepods and total zooplankton community shared a seasonal variation and an increasing trend in AA concentration from winter through spring which was associated with the spring diatom bloom. The lineal relationship observed between AA concentration in anchoveta eggs, chlorophyll a and Sea Surface Temperature (SST) suggests that the increase in phytoplankton abundance could also increase the amount of AA in the spawning female anchoveta incorporated through tissue, thus increasing the concentration in their eggs. Ascorbic acid concentrations in copepods presented size (weight) dependence. Small copepods (e.g. Acartia, Oithona) had AA concentrations two orders of magnitude higher than the heavier weight class copepods (e.g. Calanus, Rhincalanus). Results of the determination of glutathione and the antioxidant potential showed a similar trend in interannual variations, suggesting that cold SST conditions observed in the 2007 spawning season could increase the consumption of antioxidants in early stages. Potential connections between AA concentration in the food web on anchoveta reproduction and egg hatching and embryo malformations are discussed.     

Adenosine triphosphate and adenylate energy charge in marine sediments

The determination of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) concentrations in sediments from both Puget Sound and the Washington State continental shelf (USA) has been investigated. Major losses during the extraction procedure included sediment sorption, coprecipitation, and unreactivity of the soluble adenylates presumably through irreversible binding to solubilized sediment organics. Addition of phosphoric acid and the non-ionic detergent Triton X-100 to cold sulfuric acid significantly increased the extraction efficiency and sensitivity over that of sulfuric acid alone, although both methods yielded similar results when corrected for losses. Vertical profiles of ATP, total adenylates (A_T), and adenylate energy charge (EC) in sediments from both areas are presented. Profiles of A_T are similar in both areas, but EC values in the shelf sediments are significantly lower, which suggests a depressed physiological condition in the shelf population relative to that in Puget Sound.Contribution No. 513 from the University of Washington's College of Fisheries, and Contribution No. 1135 from the University of Washington's Department of Oceanography     

Distribution of lipopolysaccharide,an indicator of bacterial biomass,in subtropical areas of the sea

Lipopolysaccharide (LPS) concentrations, an indicator of bacterial biomass, were determined in the South China Sea and the Pacific Ocean. The distribution patterns of LPS were compared with those of chlorophyll a (Chl a), zooplankton biomass and the concentrations of several nutrients. LPS and total bacterial numbers in seawater were correlated with each other with a correlation coefficient (r) of 0.84 at Stations 7, 8 and 10. Diurnal fluctuation of LPS was negligible, but Chl a varied slightly in the vertical water column. Zooplankton stayed at a depth of around 400 m during the daytime and ascended quickly to the surface (0–50 m) early in the evening. The profiles of LPS and Chl a were negatively correlated to each other in the water layers above the Chl a maximum peak (r=-0.74; excluding the samples from 75 m at Station 7 and 10 m at Station 11 due to inadequate data for the statistical analysis). LPS and zooplankton biomass during the night-time, in contrast, paralleled each other at 5 stations surveyed (r=0.71). The presence of zooplankton resulted in an increase in bacterial numbers in the seawater in vitro. Based on these results, the factors controlling the occurrence and abundance of bacteria and phyto- and zooplankton in the pelagic sea are discussed.