Coping with iron limitation: a metabolomic study of <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803

Iron (Fe) is a key element for all living systems, especially for photosynthetic organisms because of its important role in the photosynthetic electron transport chain. Fe limitation in cyanobacteria leads to several physiological and morphological changes. However, the overall metabolic responses to Fe limitation are still poorly understood. In this study, we integrated elemental, stoichiometric, macromolecular, and metabolomic data to shed light on the responses of Synechocystis sp. PCC 6803, a non-N₂-fixing freshwater cyanobacterium, to Fe limitation. Compared to Synechocystis growing at nutrient replete conditions, Fe-limited cultures had lower growth rates and amounts of chlorophyll a, RNA, RNA:DNA, C, N, and P, and higher ratios of protein:RNA, C:N, C:P, and N:P, in accordance with the growth rate hypothesis which predicts faster growing organisms will have decreased biomass RNA contents and C:P and N:P ratios. Fe-limited Synechocystis had lower amounts Fe, Mn, and Mo, and higher amount of Cu. Several changes in amino acids of cultures growing under Fe limitation suggest nitrogen limitation. In addition, we found substantial increases in stress-related metabolites in Fe-limited cyanobacteria such antioxidants. This study represents an advance in understanding the stoichiometric, macromolecular, and metabolic strategies that cyanobacteria use to cope with Fe limitation. This information, moreover, may further understanding of changes in cyanobacterial functions under scenarios of Fe limitation in aquatic ecosystems.     

Effects of N and P supply on phytoplankton in Bizerte Lagoon (western Mediterranean)

Enriched bottle experiments were conducted in situ during winter (January and February) and summer (July and August) 2001 to examine the effects of nutrient enrichments (+ N, + P and + NP) on phytoplankton in Bizerte Lagoon, Tunisia. Chlorophyll a (Chl a), ranging from 3.05 μg L⁻¹ in winter to 4.52 μg L⁻¹ in summer, was dominated by the small size-faction (<5 μm) during both seasons. However, the contribution of the large size-fraction (5-200 μm) to Chl a increased from winter (26%) to summer (37%). Similarly, the carbon biomass of the 5-200 μm algae increased during the July/August period that was characterised by the high proliferation of several diatom taxa. In winter, N was the limiting element for phytoplankton growth. Its addition alone (+ N) or with P (+ NP) increased both the <5 μm and 5-200 μm Chl a concentrations. There was no change in the phytoplankton size structure, with the small cells dominating the final algal biomass in all treatments after 5 days. In summer, N and P limited the phytoplankton, but small and large algae exhibited diverse responses to different nutrient enrichments: addition of P increased the Chl a only in the 5-200 μm fraction, the + N treatment enhanced both size classes, and the NP fertilisation mostly stimulated the biomass of large cells. Consequently, the N and P addition in summer was followed by a significant change in the phytoplankton size structure, since both size-fractions contributed equally to the final Chl a biomass. Within the 5-200 μm algal community, various taxa had diverse responses to the nutrient supply during both seasons, leading to a change in the final community composition. The autotrophic flagellates appeared to grow well under N-deficient conditions. In contrast, diatom growth and biomass were mostly stimulated by the N enrichment while dinoflagellates exhibited the highest increase in their growth and biomass with P fertilisation. Our results suggest that the increasing anthropogenic supply of nutrients in the lagoon may influence algal dynamics as well as productivity in different ways depending on the nutrient composition.     

Controlling Cyanobacterial Blooms in Hypertrophic Lake Taihu,China: Will Nitrogen Reductions Cause Replacement of Non-N2 Fixing by N2 Fixing Taxa?

Excessive anthropogenic nitrogen (N) and phosphorus (P) inputs have caused an alarming increase in harmful cyanobacterial blooms, threatening sustainability of lakes and reservoirs worldwide. Hypertrophic Lake Taihu, China’s third largest freshwater lake, typifies this predicament, with toxic blooms of the non-N₂ fixing cyanobacteria Microcystis spp. dominating from spring through fall. Previous studies indicate N and P reductions are needed to reduce bloom magnitude and duration. However, N reductions may encourage replacement of non-N₂ fixing with N₂ fixing cyanobacteria. This potentially counterproductive scenario was evaluated using replicate, large (1000 L), in-lake mesocosms during summer bloom periods. N+P additions led to maximum phytoplankton production. Phosphorus enrichment, which promoted N limitation, resulted in increases in N₂ fixing taxa (Anabaena spp.), but it did not lead to significant replacement of non-N₂ fixing with N₂ fixing cyanobacteria, and N₂ fixation rates remained ecologically insignificant. Furthermore, P enrichment failed to increase phytoplankton production relative to controls, indicating that N was the most limiting nutrient throughout this period. We propose that Microcystis spp. and other non-N₂ fixing genera can maintain dominance in this shallow, highly turbid, nutrient-enriched lake by outcompeting N₂ fixing taxa for existing sources of N and P stored and cycled in the lake. To bring Taihu and other hypertrophic systems below the bloom threshold, both N and P reductions will be needed until the legacy of high N and P loading and sediment nutrient storage in these systems is depleted. At that point, a more exclusive focus on P reductions may be feasible.     

Analysis of environmental drivers influencing interspecific variations and associations among bloom-forming cyanobacteria in large,shallow eutrophic lakes

Non-diazotrophic Microcystis and filamentous N₂-fixing Aphanizomenon and Dolichospermum (formerly Anabaena) co-occur or successively dominate freshwaters globally. Previous studies indicate that dual nitrogen (N) and phosphorus (P) reduction is needed to control cyanobacterial blooms; however, N limitation may cause replacement of non-N₂-fixing by N₂-fixing taxa. To evaluate potentially counterproductive scenarios, the effects of temperature, nutrients, and zooplankton on the spatio-temporal variations of cyanobacteria were investigated in three large, shallow eutrophic lakes in China. The results illustrate that the community composition of cyanobacteria is primarily driven by physical factors and the zooplankton community, and their interactions. Niche differentiation between Microcystis and two N₂-fixing taxa in Lake Taihu and Lake Chaohu was observed, whereas small temperature fluctuations in Lake Dianchi supported co-dominance. Through structural equation modelling, predictor variables were aggregated into ‘composites’ representing their combined effects on species-specific biomass. The model results showed that Microcystis biomass was affected by water temperature and P concentrations across the studied lakes. The biomass of two filamentous taxa, by contrast, exhibited lake-specific responses. Understanding of driving forces of the succession and competition among bloom-forming cyanobacteria will help to guide lake restoration in the context of climate warming and N:P stoichiometry imbalances.     

Responses of phytoplankton to fish predation and nutrient loading in shallow lakes: a pan-European mesocosm experiment

1. The impacts of nutrients (phosphorus and nitrogen) and planktivorous fish on phytoplankton composition and biomass were studied in six shallow, macrophyte‐dominated lakes across Europe using mesocosm experiments. 2. Phytoplankton biomass was more influenced by nutrients than by densities of planktivorous fish. Nutrient addition resulted in increased algal biomass at all locations. In some experiments, a decrease was noted at the highest nutrient loadings, corresponding to added concentrations of 1 mg L^?1 P and 10 mg L^?1 N. 3. Chlorophyll a was a more precise parameter to quantify phytoplankton biomass than algal biovolume, with lower within‐treatment variability. 4. Higher densities of planktivorous fish shifted phytoplankton composition toward smaller algae (GALD < 50 μm). High nutrient loadings selected in favour of chlorophytes and cyanobacteria, while biovolumes of diatoms and dinophytes decreased. High temperatures also may increase the contribution of cyanobacteria to total phytoplankton biovolume in shallow lakes.     

Iron deficiency increases growth and nitrogen-fixation rates of phosphorus-deficient marine cyanobacteria

Marine dinitrogen (N₂)-fixing cyanobacteria have large impacts on global biogeochemistry as they fix carbon dioxide (CO₂) and fertilize oligotrophic ocean waters with new nitrogen. Iron (Fe) and phosphorus (P) are the two most important limiting nutrients for marine biological N₂ fixation, and their availabilities vary between major ocean basins and regions. A long-standing question concerns the ability of two globally dominant N₂-fixing cyanobacteria, unicellular Crocosphaera and filamentous Trichodesmium, to maintain relatively high N₂-fixation rates in these regimes where both Fe and P are typically scarce. We show that under P-deficient conditions, cultures of these two cyanobacteria are able to grow and fix N₂ faster when Fe deficient than when Fe replete. In addition, growth affinities relative to P increase while minimum concentrations of P that support growth decrease at low Fe concentrations. In Crocosphaera, this effect is accompanied by a reduction in cell sizes and elemental quotas. Relatively high growth rates of these two biogeochemically critical cyanobacteria in low-P, low-Fe environments such as those that characterize much of the oligotrophic ocean challenge the common assumption that low Fe levels can have only negative effects on marine primary producers. The closely interdependent influence of Fe and P on N₂-fixing cyanobacteria suggests that even subtle shifts in their supply ratio in the past, present and future oceans could have large consequences for global carbon and nitrogen cycles.     

Growth response of Microcystis spp. to iron enrichment in different regions of Lake Taihu, China

Iron (Fe) is an essential micronutrient for algal growth and can be a potential limiting nutrient in aquatic system, especially regions that exhibits nitrogen (N) limitation. Using short-term nutrient addition bioassays, we evaluated the potential role that iron might play in modifying the response of Microcystis spp. to the anthropogenic phosphorus (P) and N enrichment in hypereutrophic Lake Taihu, the third largest freshwater lake in China. Three nutrient enrichment experiments involving additions of N (as NO₃ ^?) and P (as PO₄ ^3?) with and without Fe were conducted during 2009?C2010 in Meiliang Bay, a region characterized by summer cyanobacterial (Microcystis spp.) blooms, and East Taihu, a region largely free of cyanobacterial blooms and dominated by macrophytes. In Meiliang Bay, Fe addition alone did not significantly increase Microcystis spp. biomass. However, Fe addition occasionally increased the stimulatory effect of N and P additions on Microcystis spp., indicating that Fe was not a primary limiting nutrient for Microcystis spp. growth. Occasionally Fe was co-limiting with N and P in this region. In East Taihu, the addition of Fe alone significantly stimulated Microcystis spp. growth, while addition of N and/or P had no effects on growth, indicating that Fe was a primary limiting nutrient in East Taihu. The combined addition of Fe and N resulted in a growth response similar to Fe alone, while combined addition of Fe and P yielded greater biomass increases than the addition of Fe alone. This indicated that in East Taihu, N was not limiting and Fe and P supplies facilitated Microcystis spp. growth. These results reflect differential availabilities and limitations of N, P, and Fe in distinct regions of Taihu. The potential role of Fe in eutrophication dynamics of large, regionally complex lakes like Taihu requires further attention.     

The role of environmental parameters in the structure of phytoplankton assemblages and cyanobacteria toxins in two hypereutrophic lakes

We evaluated the variability of cyanotoxins, water chemistry, and cyanobacteria communities in two hypereutrophic drowned river mouth lakes (Spring Lake and Mona Lake; summer 2006) in west Michigan, USA. Even with considerable geographical and watershed similarity, local variations in nutrient concentrations and environmental factors were found to influence the differences observed in cyanobacteria assemblages and cyanotoxins levels between the two lakes. Limnothrix sp. dominated the phytoplankton community in Spring Lake (82% of biovolume) and was negatively correlated with total phosphorus (TP) concentrations. Although Spring Lake was treated with alum during the previous year, Limnothrix sp. was able to bloom in the lower P environment. In contrast, the N₂-fixing cyanobacterium, Anabaena flos-aquae, dominated the phytoplankton in Mona Lake (64% of biovolume). N₂-fixing cyanobacteria dominance in Mona Lake was correlated with higher TP lower dissolved nitrogen levels. Cylindrospermopsis raciborskii was found in both systems; however, the toxin-producing polyketide synthetase gene was not present in either population. The higher TP in Mona Lake appeared to account for the 3-fold increase in cyanobacteria biovolume. Restoration plans for both lakes should include assessments of internal loading and continued phytoplankton monitoring to track the temporal distribution of cyanobacteria species and cyanotoxin concentrations.     

Iron-Stimulated N2 Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp

In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N₂ fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl₃ and EDTA-chelated FeCl₃, enhanced N₂ fixation (nitrogenase activity), photosynthesis (CO₂ fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N₂ fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms.     

The effect of nutrients on carbon and nitrogen fixation by the UCYN-A–haptophyte symbiosis

Symbiotic relationships between phytoplankton and N₂-fixing microorganisms play a crucial role in marine ecosystems. The abundant and widespread unicellular cyanobacteria group A (UCYN-A) has recently been found to live symbiotically with a haptophyte. Here, we investigated the effect of nitrogen (N), phosphorus (P), iron (Fe) and Saharan dust additions on nitrogen (N₂) fixation and primary production by the UCYN-A–haptophyte association in the subtropical eastern North Atlantic Ocean using nifH expression analysis and stable isotope incubations combined with single-cell measurements. N₂ fixation by UCYN-A was stimulated by the addition of Fe and Saharan dust, although this was not reflected in the nifH expression. CO₂ fixation by the haptophyte was stimulated by the addition of ammonium nitrate as well as Fe and Saharan dust. Intriguingly, the single-cell analysis using nanometer scale secondary ion mass spectrometry indicates that the increased CO₂ fixation by the haptophyte in treatments without added fixed N is likely an indirect result of the positive effect of Fe and/or P on UCYN-A N₂ fixation and the transfer of N₂-derived N to the haptophyte. Our results reveal a direct linkage between the marine carbon and nitrogen cycles that is fuelled by the atmospheric deposition of dust. The comparison of single-cell rates suggests a tight coupling of nitrogen and carbon transfer that stays balanced even under changing nutrient regimes. However, it appears that the transfer of carbon from the haptophyte to UCYN-A requires a transfer of nitrogen from UCYN-A. This tight coupling indicates an obligate symbiosis of this globally important diazotrophic association.     

Nutrient addition effects on chlorophyll a,phytoplankton biomass,and heterocyte formation in Lake Erie’s central basin during 2014–2017: Insights into diazotrophic blooms in high nitrogen water

1. Phosphorus (P) usually is the primary limiting nutrient of phytoplankton biomass, but attention towards nitrogen (N) and trace nutrients, such as iron (Fe), has surfaced. Additionally, N-fixing cyanobacterial blooms have been documented to occur in N-rich, P-poor waters, which is counterintuitive from the paradigm that low N and high P promotes blooms. For example, Lake Erie's central basin has Dolichospermum blooms when nitrate concentrations are high, which raises questions about which nutrient(s) are selecting for Dolichospermum over other phytoplankton and why an N-fixer is present in high N waters?
2. We conducted a 4-year (2014–2017) study in Lake Erie's central basin to determine which nutrient (P, N, or trace nutrients such as Fe, molybdenum [Mo], and boron [B]) constrained chlorophyll concentration, phytoplankton biovolume, and nitrate assimilation using nutrient enrichment bioassays. The enriched lake water was incubated in 1-L bottles in a growth chamber programmed at light and temperatures of in situ conditions for 4–7 days. We also quantified heterocytes when N-fixing cyanobacteria were present.
3. Compared to the non-enriched control, the P-enriched (+P) treatment had significantly higher chlorophyll and phytoplankton biovolume in c. 75% of experiments. Combination enrichments of P with ammonium-N, nitrate-N, Fe, Mo, and B were compared to the +P treatment to determine secondary limitations. +P and ammonium-N and +P nitrate-N resulted in higher chlorophyll in 50% of experiments but higher phytoplankton biovolume in only 25% of experiments. These results show that P was the primary limiting nutrient, but there were times when N was secondarily limiting.
4. Chlorophyll concentration indicated N secondary limitation in half of the experiments, but biovolume indicated only N secondary limitation in 25% of the experiments. To make robust conclusions from nutrient enrichment bioassays, both chlorophyll and phytoplankton biovolume should be measured.
5. The secondary effects of Fe, Mo, and B on chlorophyll were low (<26% of experiments), and no secondary effects were observed on phytoplankton biovolume and nitrate assimilation. However, +P and Fe resulted in more chlorophyll than +P in experiments conducted during Dolichospermum blooms, and +P and B significantly increased the number of heterocytes in Dolichospermum. These results indicate that low Fe availability might select for Dolichospermum, and low B constrains heterocyte formation in the central basin of Lake Erie. Furthermore, these results could apply to other lakes with high N and low P where diazotrophic cyanobacterial blooms occur.

     

Structural Diversity of Bacterial Communities Associated with Bloom-Forming Freshwater Cyanobacteria Differs According to the Cyanobacterial Genus

The factors and processes driving cyanobacterial blooms in eutrophic freshwater ecosystems have been extensively studied in the past decade. A growing number of these studies concern the direct or indirect interactions between cyanobacteria and heterotrophic bacteria. The presence of bacteria that are directly attached or immediately adjacent to cyanobacterial cells suggests that intense nutrient exchanges occur between these microorganisms. In order to determine if there is a specific association between cyanobacteria and bacteria, we compared the bacterial community composition during two cyanobacteria blooms of Anabaena (filamentous and N₂-fixing) and Microcystis (colonial and non-N₂ fixing) that occurred successively within the same lake. Using high-throughput sequencing, we revealed a clear distinction between associated and free-living communities and between cyanobacterial genera. The interactions between cyanobacteria and bacteria appeared to be based on dissolved organic matter degradation and on N recycling, both for N₂-fixing and non N₂-fixing cyanobacteria. Thus, the genus and potentially the species of cyanobacteria and its metabolic capacities appeared to select for the bacterial community in the phycosphere.     

Response of cyanobacterial mats to nutrient and salinity changes

Cyanobacterial mats (CBM), complex assemblages of cyanobacteria, bacteria and algae, are important ecosystem components of oligotrophic marshes in limestone-based regions of the Caribbean. We conducted a mesocosm experiment and evaluated the response of CBM to factorial combinations of low, medium and high phosphorus, nitrogen and salinity. Changes in composition of the main species groups of cyanobacteria and algae, primary production, cellular nutrients and enzymatic activities were recorded as response variables. The redundancy analysis with concentrations of P, N and salinity as explanatory variables showed that the primary production of CBM and the amount of phytoplankton expressed as Chl a were best explained by concentration of P, with less significant positive effect of N and a negative effect of salinity. Abundance of green algae and Chroococcales was positively correlated with increasing concentrations of P and N and reached 27.6% and 21.9%, respectively, in high P and high N treatment at the end of experiment. N₂-fixation averaged 75 and 175 nmol C₂H₄ cm⁻² min⁻¹, at low nitrogen and medium or high P, respectively, and it was negatively correlated with nitrogen concentration and positively correlated with abundance of a group of heterocytous cyanobacteria from genus Nostoc. At low N concentrations, increasing P concentrations supported higher N₂-fixation. Activity of the alkaline phosphatase, APA, was negatively correlated with P and salinity and positively with N. We also found a significant negative correlation between the APA activity and the P content of the mat. At high P and N concentrations, the mats were impacted by grazing, had a tendency to disintegrate and become shaded out by a massive growth of phytoplankton. We confirmed an overall negative effect of nutrient increase on CBM.     

Impacts of nutrient enrichment and sediment on phytoplankton community structure in the northern Baltic Sea

A three-week mesocosm experiment was conducted in order to study the effects of bottom sediment and nutrient enrichment on phytoplankton and zooplankton community structure in the Archipelago Sea, northern Baltic Sea. The transparent polyethylene enclosures included the whole water column and varied in volume from 30 to 40 m³. There were two types of enclosures: some with natural sediment as a bottom and others with a plastic bottom. The experiment was a 2 × 2 factorial design with presence of sediment and nutrient enrichment as treatment factors. Both the sediment presence and nutrient enrichment significantly increased water nutrient concentrations and the rate of primary production. However, external nutrient enrichment and the presence of sediment stimulated the growth of different phytoplankton groups, indicating that the effect of sediment was not related to nutrient fluxes alone, but involved more complex interactions. External nutrient enrichment was primarily channelled to picoplanktonic cyanobacteria, the biomass of which increased four- to fivefold due to enrichment. The presence of sediment increased the biomass of cryptophytes, chrysophytes and prasinophytes, but decreased the biomass of N₂-fixing cyanobacteria. Zooplankton biomass increased during the experiment, but was not affected by the treatments. The study shows that sediment plays a significant role in phytoplankton dynamics, underlining the importance of including sediment in shallow-water mesocosm experiments. Handling editor: J. Padisak     

Species-specific differences in phytoplankton responses to N and P enrichments and the N:P ratio in the Archipelago Sea, northern Baltic Sea

A nutrient enrichment experiment was conducted in order to studythe role of nitrogen (N), phosphorus (P) and the N:P ratio onthe early summer phytoplankton community in the ArchipelagoSea, northern Baltic Sea. The phytoplankton community was, interms of chlorophyll a and total biomass, primarily N-limited,but the individual species varied in their responses to thenutrient supply. The recorded overall N limitation was due tofast growth responses of a few N-limited species such as thediatom Chaetoceros wighamii (Brightwell) and the mixotrophicchrysophyte Uroglena sp. Another dominating diatom, Skeletonemacostatum (Greville) Cleve was most clearly P-limited. The N:Pratio had the strongest effect on Uroglena sp., which grew exponentiallyin the enrichments with a high N:P ratio. This can be explainedby the ability of the species to feed on P-rich bacteria, whichgives it a competitive advantage in P-limited conditions. Thespecies-specific differences in the responses to the nutrientenrichments can generally be explained by differences in thespecies physiology and they were consistent with the theoryof resource competition.     

Critical N:P ratio for cyanobacteria and N2-fixing species in the large shallow temperate lakes Peipsi and Võrtsjärv, North-East Europe

In the 1990s a sharp decrease in nitrogen loading occurred in Estonian rivers, bringing about a reduction of the nitrogen-to-phosphorus ratio (N:P ratio) in the large shallow lakes, Peipsi (3,555 km², mean depth 7.1 m) and Võrtsjärv (270 km², 2.8 m). The average mass ratio of total nitrogen (TN) and total phosphorus (TP) in Võrtsjärv (45) was about twice as high as that in Peipsi (22). In Peipsi, the N₂-fixing Gloeotrichia echinulata, Aphanizomenon flos-aquae and Anabaena species prevailed in the summer phytoplankton, while in Võrtsjärv the dominant cyanobacteria were Limnothrix planktonica, L. redekei and Planktolyngbya limnetica, which cannot fix N₂; the main N₂-fixing taxa Aphanizomenon skujae and Anabaena sp. seldom gained dominance. In May–October the critical TN:TP mass ratio, below which N₂-fixing cyanobacteria (Nfix) achieved high biomasses, was ～40 in Võrtsjärv and ～30 in Peipsi. The percentages of both total cyanobacteria (CY) and Nfix (CY% and Nfix%) in Peipsi achieved their maximum values at an N:P mass ratio at or below 20 for both TN:TP and Nmin:SRP. In Võrtsjärv, the TN:TP supporting a high Nfix% was between 30 and 40 and the N_min:SRP supporting this high percentage was in the same range as that in Peipsi (<20), though the maximum Nfix% values in Võrtsjärv (69%) were much lower than in Peipsi (96%). The Nmin:SRP ratio explained 77% of the variability in Nfix% in May–October. The temperature dependence of Nfix% approximated to the maximum function type, with an upper limiting value at a certain water temperature, and this was most distinct in May–October. The critical TN:TP ratios obtained from our study (roughly 30 for Peipsi and 40 for Võrtsjärv) are much higher than the Redfield N:P mass ratio routinely considered (7). Our results represent valuable guidelines for creating effective management strategies for large shallow lakes. They provide a basis for stressing the urgent need to decrease phosphorus loading and to keep the in-lake P concentration low, and not to implement nitrogen reduction measures without a simultaneous decrease of phosphorus concentration.     

Assimilation of Diazotrophic Nitrogen into Pelagic Food Webs

The fate of diazotrophic nitrogen (N_D) fixed by planktonic cyanobacteria in pelagic food webs remains unresolved, particularly for toxic cyanophytes that are selectively avoided by most herbivorous zooplankton. Current theory suggests that N_D fixed during cyanobacterial blooms can enter planktonic food webs contemporaneously with peak bloom biomass via direct grazing of zooplankton on cyanobacteria or via the uptake of bioavailable N_D (exuded from viable cyanobacterial cells) by palatable phytoplankton or microbial consortia. Alternatively, N_D can enter planktonic food webs post-bloom following the remineralization of bloom detritus. Although the relative contribution of these processes to planktonic nutrient cycles is unknown, we hypothesized that assimilation of bioavailable N_D (e.g., nitrate, ammonium) by palatable phytoplankton and subsequent grazing by zooplankton (either during or after the cyanobacterial bloom) would be the primary pathway by which N_D was incorporated into the planktonic food web. Instead, in situ stable isotope measurements and grazing experiments clearly documented that the assimilation of N_D by zooplankton outpaced assimilation by palatable phytoplankton during a bloom of toxic Nodularia spumigena Mertens. We identified two distinct temporal phases in the trophic transfer of N_D from N. spumigena to the plankton community. The first phase was a highly dynamic transfer of N_D to zooplankton with rates that covaried with bloom biomass while bypassing other phytoplankton taxa; a trophic transfer that we infer was routed through bloom-associated bacteria. The second phase was a slowly accelerating assimilation of the dissolved-N_D pool by phytoplankton that was decoupled from contemporaneous variability in N. spumigena concentrations. These findings provide empirical evidence that N_D can be assimilated and transferred rapidly throughout natural plankton communities and yield insights into the specific processes underlying the propagation of N_D through pelagic food webs.     

An artificial neural network approach for studying phytoplankton succession

Artificial neural networks are used to model phytoplankton succession and gain insight into the relative strengths of bottom-up and top-down forces shaping seasonal patterns in phytoplankton biomass and community composition. Model comparisons indicate that patterns in chlorophyll aconcentrations response instantaneously to patterns in nutrient concentrations (phosphorous (P), nitrite and nitrate (NO₂/NO₃–N) and ammonium (NH₄–H) concentrations) and zooplankton biomass (daphnid cladocera and copepoda biomass); whereas lagged responses in an index of algal community composition are evident. A randomization approach to neural networks is employed to reveal individual and interacting contributions of nutrient concentrations and zooplankton biomass to predictions of phytoplankton biomass and community composition. The results show that patterns in chlorophyll aconcentrations are directly associated with P, NO₂/NO₃–N and daphnid cladocera biomass, as well as related to interactions between daphnid cladocera biomass, and NO₂/NO₃–N and P. Similarly, patterns in phytoplankton community composition are associated with NO₂/NO₃–N and daphnid cladocera biomass; however show contrasting patterns in nutrient– zooplankton and zooplankton–zooplankton interactions. Together, the results provide correlative evidence for the importance of nutrient limitation, zooplankton grazing and nutrient regeneration in shaping phytoplankton community dynamics. This study shows that artificial neural networks can provide a powerful tool for studying phytoplankton succession by aiding in the quantification and interpretation of the individual and interacting contributions of nutrient limitation and zooplankton herbivory on phytoplankton biomass and community composition under natural conditions.     

Grazing impact of microzooplankton on a diatom bloom in a mesocosm as estimated by pigment-specific dilution technique

To investigate the impact of microzooplankton grazing on phytoplankton bloom in coastal waters, an enclosure experiment was conducted in Saanich Inlet, Canada during the summer of 1996. Daily changes in the microzooplankton grazing rate on each phytoplankton group were investigated with the growth rates of each phytoplankton group from the beginning toward the end of bloom using the dilution technique with high-performance liquid chromatography (HPLC). On Day 1 when nitrate and iron were artificially added, chlorophyll a concentration was relatively low (4.3 μg l⁻¹) and 19′-hexanoyloxyfucoxanthin-containing prymnesiophytes were predominant in the chlorophyll biomass. However, both the synthetic rates and concentrations of 19′-hexanoyloxyfucoxanthin declined before bloom, suggesting that 19′-hexanoyloxyfucoxanthin-containing prymnesiophytes weakened. Chlorophyll a concentration peaked at 23 μg l⁻¹ on Day 4 and the bloom consisted of the small chain-forming diatoms Chaetoceros spp. (4 μm in cell diameter). Diatoms were secondary constituents in the chlorophyll biomass at the beginning of the experiment, and the growth rates of diatoms (fucoxanthin) were consistently high (>0.5 d⁻¹) until Day 3. Microzooplankton grazing rates on each phytoplankton group remarkably increased except on alloxanthin-containing cryptophytes after the nutrient enrichments, and peaked with >0.6 d⁻¹ on Day 3, indicating that >45% of the standing stock of each phytoplankton group was removed per day. Both the growth and mortality rates of alloxanthin-containing cryptophytes were relatively high (>1 and >0.5 d⁻¹, respectively) until the bloom, suggesting that a homeostatic mechanism might exist between predators and their prey. Overall, microzooplankton grazing showed a rapid response to the increase in phytoplankton abundance after the nutrient enrichments, and affected the magnitude of the bloom significantly. High grazing activity of microzooplankton contributed to an increase in the abundance of heterotrophic dinoflagellates with 7-24 μm in cell size, the fraction of large-sized (>10 μm) chlorophyll a, and stimulated the growth of larger-sized ciliates after the bloom.