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.     

Release of dissolved organic matter from natural populations of marine phytoplankton

A new method is presented for measuring the rate of release of dissolved organic matter (DOM) produced by natural populations of marine phytoplankton. The method has been field-tested using natural populations of estuarine, coastal, and oceanic phytoplankton. Problems associated with the necessity for long incubation times, high initial activity of the inorganic ¹⁴C added to the sample, and self-absorption are overcome. Improved sensitivity is obtained by utilizing a large portion of the sample filtrate, Results from sampling at different times of day and for different lengths of incubation suggest that these problems should be reexamined in terms of both the percent and rate of relase of DOM from natural populations of phytoplankton. The rate of release of DOM from natural populations of marine phytoplankton decreases seaward. Phytoplankton in Georgia (USA) estuaries release <1 to 40 mg C m^-3 day^-1, in southeastern USA coastal waters from 0 to 19 mg C m^-3 day^-1, and in the westernmost Sargasso Sea from 0 to 2 mg C m^-3 day^-1. The percent of photoassimilated carbon released as DOM increases seaward. The percents are less than 7% in the Georgia estuaries, generally less than 13% in southeastern USA surface coastal waters, generally less than 21% for these coastal waters below the surface, and less than 44% for the westernmost Sargasso Sea. The results of this study and of other studies suggest that the measurement of primary productivity in estuarine and nearshore coastal waters is not seriously in error because of the absence of measurements on the rate of release of DOM from such phytoplankton. However, the estimation of primary-productivity values for oceanic waters is underestimated in some cases by about 50%.Contribution No. 595 from the Department of Oceanography, University of Washington, Seattle, Washington, USA and Contribution No. 224 from the University of Georgia Marine Institute, Sapelo Island, Georgia, USA.Extracted in part from a dissertation submitted to the Graduate Faculty of the University of Georgia in partial fulfillment of the requirements for the degree of Doctor of Philosophy.     

Iron-mediated effects on nitrate reductase in marine phytoplankton

The potential activity of nitrate reductase was determined in uni-algal cultures in the laboratory and in natural marine phytoplankton assemblages. In the laboratory bioassays, distinct differences in nitrate reductase activity were observed in iron replete versus depleted cultures for Emiliania huxleyi, Isochrysis galbana and Tetraselmis sp. Cells from iron-depleted cultures had 15 to 50 percent lower enzyme activity than those from iron-replete cultures. Upon addition of iron, nitrate reductase activity was enhanced in depleted cells up to levels comparable to those of the replete cells. Bioassays in the northern North Sea conducted in 1993, under low iron conditions, demonstrated similar results. Upon addition of 2.5 nM iron, a distinct enhancement, to a maximum of three times, of nitrate reductase activity was observed within 32 h after addition. Therefore, iron can stimulate nitrate reductase activity. In spite of the clean techniques used, some nitrate reductase activity was always observed. Iron deficiency was shown to impair nitrate reductase activity, but it is unlikely that nitrate reduction would cease completely.     

A new method for estimating phytoplankton growth rates and carbon biomass

A new method is described for the determination of phytoplankton growth rates and carbon biomass. This procedure is easy to apply and utilizes the labeling of chlorophyll a (chl a) with ¹⁴C. Pure chl a is isolated using two-way thin-layer chromatography, and the specific activity of chl a carbon is determined. Data from laboratory cultures indicate that the specific activity of chl a carbon becomes nearly equal to that of total phytoplankton carbon in incubations lasting 6 to 12 h and can be used to calculate phytoplankton growth rates and carbon biomass. Application of the method to the phytoplankton community in an eutrophic estuary in Hawaii indicates that the cells are growing with a doubling time of about 2 d and that about 85% of the particulate carbon consists of phytoplankton carbon.     

Nitrate reductase in Peru current phytoplankton

Nitrate reductase (NR) activity was assayed by measuring the NADH-dependent formation of nitrite in phytoplankton extracts. NR specific activity increased with the nitrate concentration of the water in upwelling areas of the Peru Current. The temperature optimum for NR for natural phytoplankton was 15° to 20°C. NR activity showed diel periodicity, with maximum activity about noon and minimum activity near midnight local time. Rate of nitrate reduction in the extracts averaged only about 15% of the rate of nitrate assimilation in the whole cells.     

Une approche vers l'estimation de la production potentielle du phytoplancton par analyse des cinétiques d'induction de fluorescence

In vivo chlorophyll fluorescence is particularly interesting ot ecologists because of various concepts (biomass, productivity, physiological state) associated with it. Using a modified spectrophotofluorometer, we have studied the kinetics of fluorescence in unialgal cultures and in a natural population of marine phytoplankton. Our apparatus did not achieve satisfactory results with cell suspensions having a chlorophyll concentration less than 10 g l^-1. We have also tested a method for estimating kinetics of diluted cultures and marine phytoplankton using cells collected on glass-fibre filters. For unialgal cultures in the exponential growth phase, the method proved satisfactory, and results obtained from both cell suspensions and filters were in good agreement. However, for aged cultures (principally diatoms) and natural marine phytoplankton the method proved unsuitable. The kinetics of fluorescence induction vary according to taxonomic position of the cells, light intensity of the measuring excitation beam and productiveness of the culture medium. The importance of the kinetics of fluorescence induction for characterization of phytoplankton activity is discussed.     

Eutrophication Phenomenon with Special Reference to the Kasštela Bay

Abstract

The paper discusses the eutrophication phenomenon as a result of pollution from land-based sources with special reference to the experience gained in the Kas?tela Bay Project.

The Bay of Kas?tela is a semi-enclosed bay of a total volume of 1.4 km³. It receives a great amount of untreated waste water, both domestic and industrial.

Results of the analyses of long-term data series of dissolved oxygen, nutrients, transparency and phytoplankton have shown a continuous increase of eutrophication in the Bay. While the concentration of oxygen in the euphotic layer increases due to a higher phytoplankton productivity, in the bottom layer it decreases as a result of the activity of heterotrophic bacteria. the nitrogen/phosphorus ratio in sea water has decreased, and today is much lower than in the open sea. From the Secchi-disc data it is clear that transparency has also decreased for the last three decades. Primary production as well as phytoplankton biomass has also increased. the structure of phytoplankton community has been changed and dinoflagellate species have become dominant rather than diatoms.     

Coagulation efficiency and aggregate formation in marine phytoplankton

Flocculation of phytoplankters into large, rapidly sinking aggregates has been implicated as a mechanism of vertical transport of phytoplankton to the sea floor which could have global significance. The formation rate of phytoplankton aggregates depends on the rate at which single cells collide, which is mainly physically controlled, and on the probability of adhesion upon collision (=coagulation efficiency, stickiness), which depends on physico-chemical and biological properties of the cells. We describe here an experimental method to quantify the stickiness of phytoplankton cells and demonstrate that three species of diatoms grown in the laboratory (Phaeodactylum tricornutum, Thalassiosira pseudonana, Skeletonema costatum) are indeed significantly sticky and form aggregates upon collision. The dependency of stickiness on nutrient limitation and growth was studied in the two latter species by investigating variation in stickiness as batch cultures aged. In nutrient repleteT. pseudonana cells stickiness is very low (< 5 × 10^?3), but increases by more than two orders of magnitude as cell growth ceases and the cells become nutrient limited. Stickiness ofS. costatum cells is much less variable, and even nutrient replete cells are significantly sticky. Stickiness is highest (> 10^?1) forS. costatum cells in the transition between the exponential and the stationary growth phase. The implications for phytoplankton aggregate formation and subsequent sedimentation in the sea of these two different types of stickiness patterns are discussed.     

Aspects of a zooplankton,phytoplankton, phosphorus system

In this paper a three-dimensional dynamical system which models the three-species system made up of phytoplankton, zooplankton and organic phosphorus nutrient in a lake environment is studied. The system is part of a more general limnological model for eutrophic lakes and impoundments which has been developed by Battelle Northwest Laboratories. It is shown that this system, henceforth referred to as Z-P-P, has a phase portrait comprised of a plane portrait embedded in the three-dimensional space R³ as an “attractor”.¹ Under a small perturbation of the nutrient equation it is shown that the system is essentially a classical Volterra-Lotka system embedded in a three-dimensional phase space R³.The system derived from Z-P-P by the addition of a term to the nutrient equation which represents the organic phosphorus contribution of dying phytoplankton is also considered. The equilibria of this system are studied and what can be deduced of the phase portrait is compared with that of the above systems. It is found that these phase portraits are qualitatively indifferent to the form of the growth rate functions for zooplankton and phytoplankton provided they are monotone increasing. Some discussion about the stability of these systems is included. Throughout this paper results are interpreted in limnological terms.     

A comparative study of the glutamate dehydrogenase activity in several species of marine phytoplankton

Kinetic studies on the NADP-linked glutamate dehydrogenase (GDH) activity in 8 species of marine phytoplankton have been carried out. The pH optima, linearity of reactions, apparent energies of activation, and apparent K _m values for NADPH, NH ₄ ^>+ and -ketoglutarate were determined. Based on these data, an accurate, sensitive enzyme assay procedure is presented which was successfully tested on shipboard during a recent cruise in an upwelling region. Preliminary studies on inhibition and activation of the NADP-linked GDH activity have also been carried out. In addition to NADP-linked GDH activity, NAD-linked GDH activity was discovered in certain species of phytoplankton. The possible physiological roles of NAD-and NADP-linked GDH enzymes are discussed.Contribution No. 911 from the Department of Oceanography, University of Washington. This research was supported by the National Science Foundation Grants GA-34165 and GX-33502.     

Significance of cellular nitrate content in natural populations of marine phytoplankton growing in shipboard cultures

Phytoplankton intracellular nitrate concentrations have been monitored in a 56-h experiment on a shipboard culture of surface sea water from an upwelling region. These measurements were related to parameters of biomass (particulate nitrogen) and nitrate assimilation using the ¹⁵N isotope technique and the nitrate reducase (NR) assay. The procedure for measuring cellular nitrate concentrations is described. This parameter exhibited diurnal variations, ranging from 3.1 to 20.6 ng-at nitrate per g-at particulate nitrogen, and could be correlated positively with NR activity. Nitrogen budgets show that NR activity represents only 12% of nitrate incorporation in organic phytoplankton material when nitrate is available in the sea water. However, upon depletion of the environmental nitrate (zero uptake), NR activity can fully account for the decrease of internal nitrate. From the results, it seems that internal nitrate content is a better index of nitrate consumption by marine phytoplankton than the external concentration of nitrate-nitrogen.     

Blooms of surf-zone diatoms along the coast of the Olympic Peninsula,Washington. IV. Nitrate reductase activity in natural populations and laboratory cultures of Chaetoceros armatum and Asterionella socialis

Nitrate reductase (NR) activity in the surf-zone diatom Chaetoceros armatum T. West was studied in its natural habitat at Copalis Beach, Olympic Peninsula, Washington (USA) over a period of 18 months. Variations of enzyme activity were found, depending on the season and the time of day. General seasonal trends in NR activity were correlated with nitrate concentrations in the surf-zone. The results also indicated that several factors may limit the activity of the enzyme. This is one of the reasons why NR activity could not be correlated with environmental nitrate concentration on the basis of individual measurements at a given point in time. Enzyme inhibition due to the presence of ammonium occurred on several occasions throughout the study period, and could be detected by the NR assay. This phenomenon for the most part masked any pattern of diel periodicity in NR activity that might have been expected to exist in cells in the natural environment. Cultures of C. armatum and Asterionella socialis Lewin and Norris were also studied in the laboratory. A diel fluctuation in NR activity corresponding to the pattern shown in natural phytoplankton by other authors was exhibited in C. armatum under a light-dark regime of 8 h light – 16 h dark and a light intensity of 400 lux.     

Variations in the progress of 14C uptake as a source of error in estimates of primary production

The progress curves of ¹⁴C retention for samples of phytoplankton from the Irish Sea incubated at contrasting light intensities have been obtained by two methods. The first method (A) involved the incubation of the samples for various periods up to 6 h, while the second method (B) consisted of making a series of short-term incubations over the same 6 h period. Over this period, the cumulative uptake was tenerally less when estimated by Method A than by Method B. The difference was greater in the samples incubated at the lower light intensity, the light history of the samples having no effect on the difference. The differences has a kinetic basis, with two combinations of progress curves obtained by use of the two methods. The first combination was associated with samples collected in the early morning, while the second combination was exhibited by samples taken in the afternoon irrespective of sampling depth. In certain samples, no increase in the ¹⁴C retained by the cells as measured by Method A was observed after 4 h. The cumulative retention of ¹⁴C by the cells after 2 h was generally greater when estimated by Method A than by Method B, this situation being reversed after 4 h. This reversal indicated a change in uptake kinetics between 2 and 4 h and it is suggested that this interval represents the time necessary for the ¹⁴C to work through the cellular pool of carbon. The findings are discussed in relation to the methodology for obtaining both estimates of primary production and ¹⁴C uptake-light intensity curves for marine phytoplankton.     

Tolerance of estuarine benthic diatoms to high concentrations of ammonia,nitrite ion,nitrate ion and orthophosphate

A carbon-14 assimilation method was used to determine action spectra and photosynthesis versus irradiance (P versus I) curves of natural populations of phytoplankton and zooxanthellae from a coral reef fringing Lizard Island in the Australian Barrier Reef. The action spectra were related to the phytoplankton species composition. The curves showed shade adaptation in phytoplankton from deeper waters and in the zooxanthellae. Rates of photosynthesis of zooxanthellae were shown to be highly but variably dependent on their host organisms. Photosynthetic production by zooxanthellae was about 0.9 gC m^-2 day^-1, which is about three times higher than phytoplankton production in the waters close to the reef.     

Optimization of exergy and implications of body sizes of phytoplankton and zooplankton in an aquatic ecosystem model

Size appears to be an important parameter in ecological processes. All physiological processes vary with body size ranging from small microorganisms to higher mammals. In this model, five state variables — phosphorus, detritus, phytoplankton, zooplankton and fish are considered. We study the implications of body sizes of phytoplankton and zooplankton for total system dynamics by optimizing exergy as a goal function for system performance indicator. The rates of different sub-processes of phytoplankton and zooplankton are calculated, by means of allometric relationships of their body sizes. We run the model with different combinations of body sizes of phytoplankton and zooplankton and observe the overall biomass of phytoplankton, zooplankton and fish. The highest exergy values in different combinations of phytoplankton and zooplankton size indicate the maximum biomass of fish with relative proportions of phytoplankton and zooplankton. We also test the effect of phosphorus input conditions corresponding to oligotrophic, mesotrophic, eutrophic system on its dynamics. The average exergy to be maximized over phytoplankton and zooplankton size was computed when the system reached a steady state. Since this state is often a limit cycle, and the exergy copies this behaviour, we averaged the exergy computed for 365 days (duration of 1 year) in the stable period of the run. In mesotrophic condition, maximum fish biomass with relative proportional ratio of phytoplankton, zooplankton is recorded for phytoplankton size class 3.12 (log V μm³ volume) and zooplankton size 4 (log V μm³ volume). In oligotrophic condition the highest average exergy is obtained in between phytoplankton size 1.48 (log V μm³ volume) and zooplankton size 4 (log V μm³ volume), whereas in eutrophic condition the result shows the highest exergy in the combination of phytoplankton size 5.25 (log V μm³ volume) and zooplankton size 4 (log V μm³ volume).     

Kelp as a trophic resource for marine suspension feeders: a review of isotope-based evidence

Kelp forests are enormously productive, and they and adjacent habitats support large populations of suspension feeders. What do these suspension feeders eat? Intuitively, we might expect that kelp primary production is a key form of trophic support for these animals. Indeed, a large and growing number of studies using carbon stable isotope data, typically collected over short time periods, have asserted that detritus from kelps is an important, and in some cases the main, food source for coastal benthic suspension feeders. This view has been incorporated into several textbooks and review papers covering kelp forest ecosystems, and loss of trophic support for benthic suspension feeders is now often invoked as an ecosystem consequence of top-down or other impacts on kelp forests. More direct evidence, however, suggests that these animals mainly eat phytoplankton and, in some cases, bacteria or zooplankton. Because isotope values of pure coastal phytoplankton, uncontaminated with detritus, are difficult to obtain, present studies have largely relied on single measurements from offshore environments or from the literature, which typically reflects offshore values. We review the evidence showing that phytoplankton isotope values can, and are expected to, vary widely in coastal waters and that inshore phytoplankton may often be enriched in ¹³C compared to offshore phytoplankton. This unaccounted-for variation may have systematically biased the results of such trophic studies toward finding large contributions of kelp detritus to suspension-feeder diets. We review some key stable isotope studies and put forth evidence for alternative explanations of the isotope patterns presented. Finally, we make recommendations for future isotope studies and describe several approaches for progress in this area. New techniques, particularly flow cytometry and compound-specific stable isotope analysis, provide ways to shed light on this interesting and important ecological issue.     

Evaluation of the copepod Tigriopus californicus as a bioassay organism for the detection of chemical feeding deterrents produced by marine phytoplankton

Marine phytoplankton have been shown to use chemical feeding deterrents to reduce or inhibit zooplankton grazing. In order to screen phytoplankton species for feeding deterrent production and to isolate and identify feeding deterrent compounds, a new, rapid, and reliable laboratory bioassay was developed. This bioassay used the laboratory-reared harpacticoid copepod Tigriopus californicus and measured inhibition of feeding by measuring the fecal pellet production rate. The bioassay was capable of detecting deterrent compounds: (1) adsorbed onto ground fish food (a normally palatable food); (2) dissolved in a mixture of seawater and live Thalassiosira pseudonana cells (a species of diatom which had no feeding deterrent activity); and (3) present in live cell cultures. Method (2) was recommended for use in bioassay-guided fractionation (isolation of chemical compounds), as it was reliable, rapid, accurate, and easy to perform with large numbers of samples. The total bioassay time was < 48 h, and data collection required only a microscope. Methanolic cell extracts of several phytoplankton species were screened for feeding deterrent activity. Extracts from the diatom Phaeodactylum tricornutum and the dinoflagellate Gonyaulax grindleyi gave feeding deterrent responses, while extracts from the diatom Thalassiosira pseudonana gave no feeding deterrent responses. Live P. tricornutum cells deterred feeding at densities of 6x10⁵ cells ml^-1. This bioassay should provide a valuable tool in screening phytoplankton for feeding deterrent compounds and determining the chemical nature of these compounds.     

Diet of intertidal bivalves in the Ría de Arosa (NW Spain): evidence from stable C and N isotope analysis

We used natural abundance measurements of stable carbon and nitrogen isotopes and a short-term ¹⁵N-enrichment experiment to explore the importance of river-derived and autochthonous sources to the diet of intertidal suspension-feeding bivalves in the Ría de Arosa, an oceanic-dominated system in northwest Spain. Highly seasonal and intense phytoplankton "bloom" events occur in this estuary. However, relatively low phytoplankton standing crop and productivity are present during much of the year, suggesting that food sources other than phytoplankton must support bivalves during these periods. Bivalves (Cerastoderma edule, Tapes decussatus, Mytilus galloprovincialis) were sampled quarterly at four intertidal stations along the length of the estuary with increasing distance from the Ulla River. Muscle tissue was consistently enriched in ¹³C and ¹⁵N relative to stomach contents. The stable C isotope values of bivalves indicated that river-derived inputs were, at most, a minor component of diet. These values suggested the potential importance of benthic microalgae to bivalve diet much of the year and the incorporation of ¹⁵N label applied to the sediment surface into stomach contents and muscle tissue in a field experiment supported this conclusion. Other labeled components of the microbenthos such as bacteria could also have been ingested. ¹³C-depleted values of bivalves in May coincided with elevated offshore chlorophyll a concentrations, suggesting the increased use of a phytoplankton source during the spring. We propose that intertidal suspension-feeding bivalves in this estuary use primarily resuspended microbenthos during periods of low phytoplankton concentration, but that phytoplankton increases in relative dietary importance during bloom events.Communicated by P.W. Sammarco, Chauvin     

Phytoplankton communities in aquaculture system (integration of shrimp and seaweed)

ABSTRACT

Knowledge about phytoplankton communities is important for aquaculture due to their influence on environmental variables. The goal of the study was to evaluate the ecological effect of seaweed Gracilaria birdiae at three different densities: 2.5 (LG2.5); 5.0 (LG5.0) and 7.5?kg m^?3 (LG7.5), on water quality variables and the phytoplankton communities in integrated aquaculture systems (shrimp and seaweed) with a control (without seaweed) (LB) using biofloc technology. Settleable and total suspended solids were significantly correlated with the phytoplankton groups. In addition, orthophosphate was the main variable controlling phytoplankton growth and gross primary production. No significant differences were identified among the treatments in terms of the phytoplankton assemblages (p?>?0.05) but differences were found between the communities (ANOSIM, R global?=?0.873) in relation to the days of the experiment. However, the LG2.5 treatment had higher diversity and lower species dominance, as well as higher productivity and final shrimp weight. The results indicate that seaweed inhibit phytoplankton growth, mainly of toxic species. They suggest that phytoplankton composition in integrated aquaculture systems in the presence of seaweed at a density of 2.5?kg m^?3 is more diversified and provides better water quality and shrimp performance by inhibiting the abundance of harmful species.     

Improved fluorescent microscopy for measuring the standing stock of phytoplankton including fragile components

Fluorescent microscopy has been used to separate autotrophic from heterotrophic microorganisms in environmental studies. We report on an improvement on this method for measuring standing stock of phytoplankton. Our method employs a membrane filter and can measure numbers of living phytoplankton including fragile flagellates and size of phytoplankton cells. This method was successfully used in Tokyo Bay and Kuroshio regions.