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UVB effects on the photosystem II-D1 protein of phytoplankton and natural phytoplankton communities.

The reaction center of photosystem II is susceptible to photodamage. In particular the D1 protein located in the photosystem II core has a rapid, light-dependent turnover termed the photosystem II repair cycle that, under illumination, degrades and resynthesizes D1 protein to limit accumulation of photodamaged photosystem II. Most studies concerning the effects of UVB (280-320 nm) on this cycle have been on cyanobacteria or specific phytoplankton species rather than on natural communities of phytoplankton. During a 5-year multidisciplinary project on the effects of UV radiation (200-400 nm) on natural systems, the effects of UVB on the D1 protein of natural phytoplankton communities were assessed. This review provides an overview of photoinhibitory effects of light on cultured and natural phytoplankton, with an emphasis on the interrelation of UVB exposure, D1 protein degradation and the repair of photosystem II through D1 resynthesis. Although the UVB component of the solar spectrum contributes to the primary photoinactivation of photosystem II, we conclude that, in natural communities, inhibition of the rate of the photosystem II repair cycle is a more important influence of UVB on primary productivity. Indeed, exposing tropical and temperate phytoplankton communities to supplemented UVB had more inhibitory effect on D1 synthesis than on the D1 degradation process itself. However, the rate of net D1 damage was faster for the tropical communities, likely because of the effects of high ambient light and water temperature on mechanisms of protein degradation and synthesis.

Chloroplasts↗

Establishing boundary classes for the classification of UK marine waters using phytoplankton communities.

This paper presents a description of three of the proposed phytoplankton indices under investigation as part of a classification framework for UK and ROI marine waters. The three indices proposed for the classification process are (i) phytoplankton biomass measured as chlorophyll, (ii) the frequency of elevated phytoplankton counts measuring individual species and total cell counts and (iii) Seasonal progression of phytoplankton functional groups through the year. Phytoplankton biomass is calculated by a 90th percentile measurement of chlorophyll over the growing season (April to September) compared to a predetermined reference value. Calculation of functional groups and cell counts are taken as proportional counts derived from the presence of the indicator species or group as compared to the total phytoplankton count. Initial boundary conditions for the assessment of high/good status were tested for each index. Chlorophyll reference conditions were taken from thresholds developed for previous EU directives with the setting of offshore concentrations as a reference condition. Thresholds for elevated counts of phytoplankton taxa were taken from previous EU assessments describing counts that could be impact negatively on the environment. Reference seasonal growth curves are established using phytoplankton counts from "high status" waterbodies. To test the preliminary boundaries for each index, a risk assessment integrating nutrient enrichment and susceptibility for coastal and transitional waters was carried out to identify WFD waterbodies in England and Wales at different levels of risk. Waterbodies assessed as having low or medium risk from nutrient enrichment were identified as type 1 and type 2 waterbodies, and waterbodies assessed as high risk were identified as type 3 waterbodies. Phytoplankton data was extracted from the risk assigned waterbodies and applied to each phytoplankton index to test the robustness of the preliminary classification ranges for each phytoplankton index.

Chlorophyll↗

Adaptive evolution of phytoplankton cell size.

We present a simple nutrient-phytoplankton-zooplankton (NPZ) model that incorporates adaptive evolution and allometric relations to examine the patterns and consequences of adaptive changes in plankton body size. Assuming stable environmental conditions, the model makes the following predictions. First, phytoplankton should evolve toward small sizes typical of picoplankton. Second, in the absence of grazers, nutrient concentration is minimized as phytoplankton reach their fitness maximum. Third, increasing nutrient flux tends to increase phytoplankton cell size in the presence of phytoplankton-zooplankton coevolution but has no effect in the absence of zooplankton. Fourth, phytoplankton reach their fitness maximum in the absence of grazers, and the evolutionary nutrient-phytoplankton system has a stable equilibrium. In contrast, phytoplankton may approach their fitness minimum in the evolutionary NPZ system where phytoplankton and zooplankton are allowed to coevolve, which may result in oscillatory (unstable) dynamics of the evolutionary NPZ system, compared with the otherwise stable nonevolutionary NPZ system. These results suggest that evolutionary interactions between phytoplankton and zooplankton may have contributed to observed changes in phytoplankton sizes and associated biogeochemical cycles over geological time scales.

Adaptation, Physiological↗

Cladocerans versus copepods: the cause of contrasting top-down controls on freshwater and marine phytoplankton.

Top-down control of phytoplankton by crustacean mesozooplankton is a cornerstone of freshwater ecology. Apparently, trophic cascades are more frequently reported from freshwater than from marine plankton. We argue that this difference is real and mainly caused by biological differences at the zooplankton-phytoplankton link: cladocerans (particularly Daphnia) in the lakes and copepods in the sea. We derive these conclusions from recent literature and a number of own, similarly designed mesocosm experiments conducted in a lake, a brackish water and a marine site. In all experiments, phytoplankton were exposed to gradients of experimentally manipulated densities of zooplankton, including freshwater copepods and cladocerans, and marine copepods and appendicularians. The suggested reasons for the difference between lake and marine trophic cascades are: (1) Both copepods and cladocerans suppress only part of the phytoplankton size spectrum: cladocerans the small and copepods the large phytoplankton. (2) If not controlled by grazing, small phytoplankton may increase their biomass faster than large phytoplankton. (3) Copepods additionally release small phytoplankton from grazing pressure by intermediate consumers (protozoa) and competitors (predation on appendicularian eggs), while cladocerans do not release large phytoplankton from grazing pressure by any functional group. (4) Cladocerans sequester more of the limiting nutrient than copepods, leaving fewer nutrients available for compensatory growth of ungrazed phytoplankton.

Animals↗

Multivariate analysis of interactions between phytoplankton biomass and environmental variables in Taihu Lake, China.

Phytoplankton variation in large shallow eutrophic lakes is characterized by high spatial and temporal heterogenity. Understanding the pattern of phytoplankton variation and the relationships between it and environmental variables can contribute to eutrophic lakes management. In this study Taihu Lake, one of the largest eutrophic fresh water lake in China, was taken as study area. The water body of Taihu Lake was divided into five regions viz. Wuli bay (WB), Meilian Bay (MB), West Taihu Lake (WTL), Main Body of Taihu Lake (MBTL) and East Taihu Lake (ETL). Concentrations of chlorophyll-a and the related environmental variables were determined in each region in the period 2000-2003. Factor analysis and multivariate analysis were applied to evaluate the interactions between phytoplankton variation and environmental variables. Results showed that the highest average concentrations of TN, TP and Chl-a were observed in WB, followed in a descending order by MB and WTL, and the lowest concentrations of TN, TP and Chl-a were observed in MBTL and ETL. Chl-a and TP concentrations in most regions (except ETL) declined during the study period. It suggested that to some extent the lake was recovering from eutrophication. However, persistent ascending of TN and NH(4)-N in all five regions indicated the deteriorating of water quality in the study period. Results of multivariate showed that the relationships between phytoplankton biomass and environmental variables varied among regions. TP illustrated itself a controlling role on phytoplankton in WB, MB, WTL and MBTL according to the significant positive relations to phytoplankton biomass in these regions. Nitrogen could be identified as a limiting factor to phytoplankton biomass in ETL in view of the positive correlations between TN and phytoplankton and between NH(4)-N and phytoplankton. Spatial variation of interactions between phytoplankton and environmental parameters suggested proper eutrophication control measures were needed to restore ecological system in each region of Taihu Lake.

Biomass↗

Standing stock and production rate of phytoplankton and a red tide outbreak in a heavily eutrophic embayment, Dokai Bay, Japan.

The seasonal variation of phytoplankton biomass and primary productivity in a heavily eutrophic embayment, Dokai Bay, Japan, was determined. Dokai Bay was characterized by high phytoplankton biomass and productivity during summer and low phytoplankton biomass and productivity during other seasons. The results suggested that phytoplankton growth was limited by only irradiance and water temperature under the high nutrient concentrations available for phytoplankton growth in the entire year. Moreover, in spite of sufficient nutrient for phytoplankton growth in the entire year, a red tide occurred only in the summer period in this bay. Our results suggested that a red tide occurred by the high phytoplankton growth rate in the summer season, but in other periods surface phytoplankton was flushed out of the bay before forming the red tide, because phytoplankton growth rate was low and could not form the red tide due to low irradiance and low water temperature.

Animals↗

Impact of trace metals and nutrients levels on phytoplankton from the Kuwait coast.

The impact of marine pollution was estimated from seven stations of Kuwait coastal waters. The mean concentrations of all trace metals in seawater (0.07-7.04 microg/l) and in phytoplankton (14.90-52.01 microg/g) revealed less phytoplankton abundance in Stations IV VI during the four seasons. However, a relative surge in the phytoplankton abundance, with an increase in the mean concentrations of all trace metals in seawater (0.15-8.94 microg/l) and phytoplankton (22.82-61.25 microg/g), was observed in Station II. A low phytoplankton abundance was noted in Stations I, III, and VII despite low mean concentrations of all the trace metals in seawater (0.11-5.77 microg/l) and phytoplankton (1.40-20 microg/g) during the four seasons. Statistical tests revealed significant difference in Fe, Ni, and Pb between the four seasons in seawater and Fe and Co in phytoplankton. No significant difference in trace metals between locations was observed in seawater and in phytoplankton. The major nutrients in all the four seasons were found in the magnitude of SiO3 > NO3 > PO4 in all the stations, and thus, support the presence of rich diatoms and dinoflagellates than other phytoplankton groups. The seven stations also represented indicator species of oil and industrial waste pollution.

Animals↗

Phytoplankton community and chlorophyll a as trophic state indices of Lake Skadar (Montenegro, Balkan).

BACKGROUND, AIMS AND SCOPE: Phytoplankton, as a first step in trophic cascades of lakes, can be a good indicator of trophic states, considering that every environmental change affects this community and many species of this community are sensitive to changes, and that they response very quickly. In this study, we tried to assess and predict the trophic state of Lake Skadar according to phytoplankton data. METHODS: Water samples were collected using Ruttner sampling bottle. Temperature, dissolved oxygen, ph, conductivity and transparence were measured in situ using portable equipment. Nutrients and chlorophyll a were measured using standard spectrophotometric methods. A determination of phytoplankton species was performed using relevant keys and the counting of cells was performed using sedimentation methods. RESULTS AND DISCUSSION: The species composition of Lake Skadar revealed 95 taxa, with Chlorophyceae and Bacillariophyceae being represented best. According to an average chlorophyll a concentration of 5.9 pg/l, Lake Skadar belongs to the mesotrophic level of the trophic scale. Developed prediction equation for chlorophyll a revealed a good prediction (R2 = 0.71) and the parameter Secchi depth was primarily correlated with chlorophyll a concentration. Trophic state indices derived from chlorophyll a and transparency, were close together, but both were below the phosphorous index. Values of trophic state indices rank the Lake Skadar as being mesotrophic. This study also showed that indices of diversity based on phytoplankton are weak indicators of trophic status and that they can well characterize only differences between assemblages and associations. According to calculated saprobic indices (ranging from 1.5 to 2.15), Lake Skadar is on betamesosaprobic level of saprobity, which means that it is moderately polluted with organic compounds. CONCLUSIONS: Total phosphorus is not the main limiting factor for the phytoplankton community in Lake Skadar. Disagreements between chlorophyll and the transparency index, on the one hand, and the total phosphorus index, on the other, suggest that the phytoplankton in Lake Skadar is probably limited by other factors than phosphorus, such as nitrogen, toxic substances or intense zooplankton grazing. According to the majority of investigated parameters and indices derived from phytoplankton data, Lake Skadar is mesotrophic, with tendencies toward eutrophic levels during the summer period. RECOMMENDATIONS AND OUTLOOK: Long-term monitoring is required for a better estimation of state and the conditions of Lake Skadar. Further studies on factors influencing the phytoplankton community, especially zooplankton grazing and toxic substances, which were not included in this study, should be continued in the future to improve the efficiency of phytoplankton usage in estimating the ecological and trophic conditions of Lake Skadar.

Albania↗

[Distribution of phytoplankton and water dynamical environmental factors in high red tide occurrence area of Changjiang River estuary].

Species composition and cell abundance of phytoplankton, and its relations with environmental factors were investigated in the Changjiang River estuary of east China sea (30 degrees 50'-31 degrees 50'N, 121 degrees 50'-123 degrees 00'E). A total of 110 taxa belonging to 45 genera of phytoplankton in the investigation area were identified. The main species of phytoplankton resulted in red tide were Skeletonema costatum and Prorocentrum dentatum, etc. Phytoplankton abundance in the day was higher than that at night. The vertical distribution of phytoplankton in different water bodies was not obvious. The ecological groups of phytoplankton were freshwater group, estuarine and brackish water and low salinity group in the coast, high salinity and warm water group in off-sea, and widely ranging group. The abundance of phytoplankton in the Changjiang River estuary was 1.6 x 10(3)-75.2 x 10(3) ind.dm-3. The species composition and cell abundance of phytoplankton were related to the freshwater pouring into the Changjiang River estuary. The three waters in the area affecting the distribution of phytoplankton were the Changjiang River estuary water, freshwater, and warm water from outsea.

Ecosystem↗

Food web structure in the recently flooded Sep Reservoir as inferred from phytoplankton population dynamics and living microbial biomass.

Phytoplankton dynamics, bacterial standing stocks and living microbial biomass (derived from ATP measurements, 0.7-200 mm size class) were examined in 1996 in the newly flooded (1995) Sep Reservoir ('Massif Central,' France), for evidence of the importance of the microbial food web relative to the traditional food chain. Phosphate concentrations were low, N:P ratios were high, and phosphate losses converted into carbon accounted for <50% of phytoplankton biomass and production, indicating that P was limiting phytoplankton development during the study. The observed low availability of P contrasts with the high release of "directly" assimilable P often reported in newly flooded reservoirs, suggesting that factors determining nutrient dynamics in such ecosystems are complex. The phosphate availability, but also the water column stability, seemed to be among the major factors determining phytoplankton dynamics, as (i) large-size phytoplankton species were prominent during the period of increasing water column stability, whereas small-size species dominated phytoplankton assemblages during the period of decreasing stability, and (ii) a Dinobryon divergens bloom occurred during a period when inorganic P was undetectable, coinciding with the lowest values of bacterial standing stocks. Indication of grazing limitation of bacterial populations by the mixotrophic chrysophyte D. divergens (in late spring) and by other potential grazers (mainly rotifers in summer) seemed to be confirmed by the Model II or functional slopes of the bacterial vs phytoplankton regressions, which were always <0.63. Phytoplankton biomass was not correlated with phosphorus sources and its contribution was remarkably low relative to the living microbial biomass which, in contrast, was positively correlated with total phosphorus in summer. We conclude that planktonic microheterotrophs are strongly implicated in the phosphorus dynamics in the Sep Reservoir, and thus support the idea that an important amount of matter and energy flows through the "microbial loop" and food web, shortly after the flooding of a reservoir.

Bacteria↗

Links between phytoplankton and bacterial community dynamics in a coastal marine environment.

Bacteria and phytoplankton dynamics are thought to be closely linked in coastal marine environments, with correlations frequently observed between bacterial and phytoplankton biomass. In contrast, little is known about how these communities interact with each other at the species composition level. The purpose of the current study was to analyze bacterial community dynamics in a productive, coastal ecosystem and to determine whether they were related to phytoplankton community dynamics. Near-surface seawater samples were collected in February, May, July, and September 2000 from several stations in the Bay of Fundy. Savin et al. (M.C. Savin et al., Microb Ecol 48: 51-65) analyzed the phytoplankton community in simultaneously collected samples. The attached and free-living bacterial communities were collected by successive filtration onto 5 microm and 0.22 microm pore-size filters, respectively. DNA was extracted from filters and bacterial 16S rRNA gene fragments were amplified and analyzed by denaturing gradient gel electrophoresis (DGGE). DGGE revealed that diversity and temporal variability were lower in the free-living than the attached bacterial community. Both attached and free-living communities were dominated by members of the Roseobacter and Cytophaga groups. Correspondence analysis (CA) ordination diagrams showed similar patterns for the phytoplankton and attached bacterial communities, indicating that shifts in the species composition of these communities were linked. Similarly, canonical CA revealed that the diversity, abundance, and percentage of diatoms in the phytoplankton community accounted for a significant amount of the variability in the attached bacterial community composition. In contrast, ordination analyses did not reveal an association between free-living bacteria and phytoplankton. These results suggest that there are specific interactions between phytoplankton and the bacteria attached to them, and that these interactions influence the composition of both communities.

Atlantic Ocean↗

The impact of diel vertical migration of Daphnia on phytoplankton dynamics.

Diel vertical migration (DVM) of large zooplankton is a very common phenomenon in the pelagic zone of lakes and oceans. Although the underlying mechanisms of DVM are well understood, we lack experimental studies on the consequences of this behaviour for the zooplankton's food resource-the phytoplankton. As large zooplankton species or individuals migrate downwards into lower and darker water strata by day and upwards into surface layers by night, a huge amount of herbivorous biomass moves through the water column twice a day. This migration must have profound consequences for the phytoplankton. It is generally assumed that migration supports an enhanced phytoplankton biomass and a change in the composition of the phytoplankton community towards smaller, edible algae in the epilimnion of a lake. We tested this assumption for the first time in field experiments by comparing phytoplankton biomass and community assemblage in mesocosms with and without artificially migrating natural stocks of Daphnia hyalina. We show that DVM can enhance phytoplankton biomass in the epilimnion and that it has a strong impact on the composition of a phytoplankton community leading to an advantage for small, edible algae. Our results support the idea that DVM of Daphnia can have strong effects on phytoplankton dynamics in a lake.

Animals↗

Interactive effects of light and nutrients on phytoplankton stoichiometry.

The stoichiometric composition of autotrophs can vary greatly in response to variation in light and nutrient availability, and can mediate ecological processes such as C sequestration, growth of herbivores, and nutrient cycling. We investigated light and nutrient effects on phytoplankton stoichiometry, employing five experiments on intact phytoplankton assemblages from three lakes varying in productivity and species composition. Each experiment employed two nutrient and eight irradiance levels in a fully factorial design. Light and nutrients interactively affected phytoplankton stoichiometry. Thus, phytoplankton C:N, C:P, and N:P ratios increased with irradiance, and slopes of the stoichiometric ratio versus irradiance relationships were steeper with ambient nutrients than with nutrients added. Our results support the light-nutrient hypothesis, which predicts that phytoplankton C:nutrient ratios are functions of the ratio of available light and nutrients; however, we observed considerable variation among lakes in the expression of this relationship. Phytoplankton species diversity was positively correlated with the slopes of the C:N and C:P versus irradiance relationships, suggesting that diverse assemblages may exhibit greater flexibility in the response of phytoplankton nutrient stoichiometry to light and nutrients. The interactive nature of light and nutrient effects may render it difficult to generate predictive models of stoichiometric responses to these two factors. Our results point to the need for future studies that examine stoichiometric responses across a wide range of phytoplankton communities.

Animals↗

Patterns and mechanisms of phytoplankton variability in Lake Washington (USA).

Temporal variability in lake phytoplankton is controlled largely by a complex interplay between hydrodynamic and chemical factors, and food web interactions. We explored mechanisms underlying phytoplankton interannual variability in Lake Washington (USA), using a 25-yr time series of water quality data (1975-1999). Time-series analysis and PCA were used to decompose chlorophyll data into modes of variability. We found that phytoplankton dynamics in Lake Washington were characterized by four seasonal modes, each of which was associated with different ecological processes. The first mode coincided with the period when the system was light limited (January-March) and phytoplankton patterns were driven by the amount of available solar radiation. The second mode (April-June) coincided with the peak of the spring bloom and the subsequent decline of phytoplankton biomass, and was largely controlled by total phosphorus levels and grazing pressure from cladoceran zooplankton. Evidence of co-dependence and tight relationship between phytoplankton and cladoceran dynamics were also found from July to October when a large portion of the phosphorus supply in the mixed layer was provided by zooplankton excretion. The fourth mode (November-December) was associated with the transition to thermal and chemical homogeneity and the winter phytoplankton minima (2-2.5 microg/l). Finally, we examined the effects of meteorological forcing and large-scale oceanic climate fluctuations (ENSO and PDO) on phytoplankton dynamics and assessed the significance of their role on the interannual variability in the lake.

Animals↗

Phylogenetic diversity and specificity of bacteria closely associated with Alexandrium spp. and other phytoplankton.

While several studies have suggested that bacterium-phytoplankton interactions have the potential to dramatically influence harmful algal bloom dynamics, little is known about how bacteria and phytoplankton communities interact at the species composition level. The objective of the current study was to determine whether there are specific associations between diverse phytoplankton and the bacteria that co-occur with them. We determined the phylogenetic diversity of bacterial assemblages associated with 10 Alexandrium strains and representatives of the major taxonomic groups of phytoplankton in the Gulf of Maine. For this analysis we chose xenic phytoplankton cultures that (i) represented a broad taxonomic range, (ii) represented a broad geographic range for Alexandrium spp. isolates, (iii) grew under similar cultivation conditions, (iv) had a minimal length of time since the original isolation, and (v) had been isolated from a vegetative phytoplankton cell. 16S rRNA gene fragments of most Bacteria were amplified from DNA extracted from cultures and were analyzed by denaturing gradient gel electrophoresis and sequencing. A greater number of bacterial species were shared by different Alexandrium cultures, regardless of the geographic origin, than by Alexandrium species and nontoxic phytoplankton from the Gulf of Maine. In particular, members of the Roseobacter clade showed a higher degree of association with Alexandrium than with other bacterial groups, and many sequences matched sequences reported to be associated with other toxic dinoflagellates. These results provide evidence for specificity in bacterium-phytoplankton associations.

Animals↗

Seasonal study on the Cd, Se, and Zn uptake by natural coastal phytoplankton assemblages.

We conducted a seasonal study of the uptake of Cd, Se(IV), and Zn by natural phytoplankton assemblages collected from two locations in Hong Kong coastal waters. Marine phytoplankton in Port Shelter was dominated by diatoms and cyanobacteria, while phytoplankton in Tolo Harbour was dominated by dinoflagellates. The natural phytoplankton assemblages were spiked with macronutrients (N and P) and metals, and the accumulation of these metals was followed for 3 to 5 d in the laboratory. The two coastal systems were mostly P limited, with N limitation observed only during the summer in Tolo Harbour. With the additions of these macronutrients, phytoplankton responded in different ways in their accumulation of trace metals. Cadmium concentrations (normalized to chlorophyll a [chl a] concentration) increased by 25 to 38% with the simultaneous addition of N and P but were roughly constant when N alone was added. In N-limited systems, the Cd concentration increased by 25% with N addition. Concentrations of Se were reduced competitively (by 40-45%) with P addition. Zinc concentrations were reduced by 23 to 38% when both N and P were added, largely because of a major increase in biomass (biodilution). Both Cd and Zn uptake rates increased with increasing growth rates. The uptake rate of Se was not related to the growth rate because of the confounding influence of P addition. The particle concentration effect was observed only for Zn, in which the increasing chl a concentration biodiluted the Zn concentration in the phytoplankton. Overall, our results suggest that macronutrients may affect trace metal accumulation in natural phytoplankton because of their controls on cell growth, which may have implications for our understanding of trace metal dynamics in coastal ecosystems during phytoplankton blooms as well as for the prediction of trophic transfer in food chains.

Cadmium↗

Uncoupling of bacterioplankton and phytoplankton production in fresh waters is affected by inorganic nutrient limitation.

Pelagic bacterial production is often positively correlated, or coupled, with primary production through utilization of autotrophically produced dissolved organic carbon. Recent studies indicate that inorganic N or P can directly limit both bacterial and phytoplanktonic growth. Our mesocosm experiments, with whole communities from mesotrophic Calder Lake, test whether this apparent bacterial-algal coupling may be the result of independent responses to limiting inorganic nutrients. In systems without N additions, numbers of bacteria but not phytoplankton increased 2- to 2.5-fold in response to P fertilization (0 to 2.0 mumol of P per liter); this resulted in uncoupled production patterns. In systems supplemented with 10 mumol of NH(4)NO(3) per liter, P addition resulted in up to threefold increases in bacteria and two- to fivefold increases in total phytoplankton biomass (close coupling). P limitation of pelagic bacteria occurred independently of phytoplankton dynamics, and regressions between bacterial abundance and phytoplankton chlorophyll a were nonsignificant in all systems without added N. We describe a useful and simple coupling index which predicts that shifts in phytoplankton and bacterioplankton growth will be unrelated (Delta bacteria/Delta phytoplankton --> either + infinity or - infinity) in systems with inorganic N/P (molar) ratios of < approximately 40. In systems with higher N/P ratios (>40), the coupling index will approach 1.0 and close coupling between bacteria and phytoplankton is predicted to occur.

Journal Article↗

Phytoplankton monitoring by high performance flow cytometry: a successful approach?

BACKGROUND: Regular phytoplankton monitoring in Dutch coastal waters is performed as an indicator of the ecological state of these waters. The monitoring program is focused on temporal and spatial changes of species composition and abundance. Flow cytometry has been introduced to provide additional information, to improve ecosystem understanding, and to increase the efficiency of analysis and reportage. METHODS: Phytoplankton community abundance and composition were routinely determined by flow cytometry and microscopy at six locations in the North Sea over three annual cycles between 2000 and 2003. Supplementary measurements were also made for fluorescence (chlorophyll-a and other pigments) and, in combination with flow cytometric and microscopic data, were used to determine phytoplankton abundance and composition as a function of their size distribution. Real-time imaging of species was also used to identify species on the basis of their flow cytometric optical characteristics. RESULTS: Flow cytometric analysis took 15 min on average. Analysis including data processing, and Web site reportage took less than 1 h. Phytoplankton concentrations (cells/ml), biomass (fluorescence/ml), and concentration of phycoerythrin- or phycocyanin-containing cells (cells/ml) as a function of their algal size were produced every 2 weeks on average. The phytoplankton integrated annual concentration and biomass were used as ecological indicators for overall phytoplankton status. Real-time imaging of cells in flow enabled the identification of dominant species and was applied as an early warning system for Phaeocystis spp. CONCLUSIONS: The reproducibility and count precision due to the large number of observations of the flow cytometric technique provided reliable data for monitoring long-term trends. Flow cytometrically based analyses extended the lower detection limit (<0.5 microm) of analysis beyond the capabilities of other techniques such as the relation between small and larger phytoplankton, the relation between cell counts and biomass as a function of cell size, but also the ability to monitor and report on blooms of harmful algae. A good correlation was found between concentrations (cells/ml) measured by flow cytometry and microscopy. In practice, flow cytometric analysis of a single marine sample took 15 min on average.

Animals↗