Effects of the diatom-Emiliana huxleyi succession on photosynthesis, calcification and carbon metabolism by size-fractioned phytoplankton

Changes in the species composition, photosynthesis, calcification and size-fractionated carbon metabolism by natural phytoplankton assemblages were monitored in three mesocosms under different nutrient conditions during May 1993. In the 3 enclosures, the decline of the diatom-dominated assemblages was followed by the development of a bloom of the coccolithoporid Emiliania huxleyi. Highest growth of E. huxleyi was observed in the mesocosm with a high N : P ratio, suggesting this species is a good competitor at low phosphate concentrations. The transition from diatom- to E. huxleyi-dominated assemblages brought about a sharp reduction of the phytoplankton standing stock and carbon-specific photosynthetic rate. The relative contribution of the smaller size fraction to total photosynthesis increased as the succession progressed. Calcification rate and E. huxleyi cell-specified calcite production were highest during the early stages of development of the E. huxleyi bloom. Distinct changes in the patterns of ¹⁴C allocation into biomolecules were noticed during the diatom-E. huxleyi succession. The diatom-dominated assemblage showed high relative ¹⁴C incorporation into low molecular weight metabolites (LMWM), whereas proteins and, specially, lipids accounted for the largest proportion of carbon incorporation in the E. huxleyi bloom. The patterns of photoassimilated carbon metabolism proved to be strongly dependent on cellular size, as protein relative synthesis was significantly higher in the smaller than in the larger size fraction, irrespective of the nutrient regime and the successional stage. These results are discussed in relation to the ecological and physiological features of small phytoplankton.     

Production and decomposition of new DOC by marine plankton communities: carbohydrates, refractory components and nutrient limitation

The accumulation and biodegradation of dissolved organic carbon (DOC) and dissolved and particulate combined neutral sugars (DCNS, PCNS) were followed during a period of 22 days in experimental marine phytoplankton incubations. Five different growth regimes were established in 11 m³ coastal mesocosms to test whether an alternate Redfield ratio with either N or P depletion and ±diatom dominance would induce accumulation of refractory DOC (RDOC) and DCNS. The highest accumulation of DOC, DCNS and PCNS was found in the diatom dominated mesocosms. Sixteen percent of the newly accumulated DOC in the mesocosms with diatoms dominating could be explained by DCNS, while only 6% was explained in the mesocosms with few diatoms. PCNS composition was similar in all mesocosms and with dominance of glucose and mannose, while DCNS were more evenly distributed with the following mole percentages fucose 15, rhamnose 14, arabinose 6, galactose 27, glucose 20 and mannose 18%. The DCNS composition did not reflect the PCNS composition at any time during the experiment. Accumulated DCNS were quickly degraded and only 1% of the new RDOC was explained by DCNS. RDOC accumulated after day # 17 in the two mesocosms driven towards the most severe P limitation both with and without silicate. This shows that RDOC can be produced directly by the phytoplankton or indirectly in food web processes during the later stages of a bloom where the phytoplankton is P limited.     

Community Dynamics of Free-living and Particle-associated Bacterial Assemblages during a Freshwater Phytoplankton Bloom

Bacterial community dynamics were followed in a 19-day period during an induced diatom bloom in two freshwater mesocosms. The main goal was to compare diversity and succession among free-living (<10 MM) AND PARTICLE-ASSOCIATED (>10 mm) bacteria. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified 16S rDNA showed the highest number of bands among free-living bacteria, but with a significant phylogenetic overlap in the two size fractions indicating that free-living bacteria were also important members of the particle-associated bacterial assemblage. Whereas the number of bands in the free-living fraction decreased during the course of the bloom, several phylotypes unique to particles appeared towards the end of the experiment. Besides the primer set targeting Bacteria, a primer set targeting most members of the Cytophaga-Flavobacterium (CF)-cluster of the Cytophaga-Flavobacterium-Bacteroides group and a primer set mainly targeting a-Proteobacteria were applied. PCR-DGGE analyses revealed that a number of phylotypes targeted by those primer sets were found solely on particles. Almost all sequenced bands from the bacterial DGGE gel were related to phylogenetic groups commonly found in freshwater: a-Proteobacteria, CF, and Firmicutes. Despite the use of primers intended to be specific mainly for a-Proteobacteria most bands sequenced from the a-proteobacterial DGGE gel formed a cluster within the Verrucomicrobiales subdivision of the Verrucomicrobia division and were not related to a-Proteobacteria. Bands sequenced from the CF DGGE gel were related to members of the CF cluster. From the present study, we suggest that free-living and particle-associated bacterial communities should not be perceived as separate entities, but rather as interacting assemblages, where the extent of phylogenetic overlap is dependent on the nature of the particulate matter.     

Phaeocystis globosa and the phytoplankton succession in the East Frisian coastal waters

The phytoplankton succession and related abiotic variables were investigated within the frame of a monitoring programme. Results from 1985 to 1991 are presented. For the prymnesiophytePhaeocystis globosa, an increase in the maximum annual abundance was observed. The appearance of this species in the phytoplankton succession followed an annually recurrent pattern. Among the starting conditions for the bloom, a coincidence with the annual DIN/P maximum was most obvious. Quantitatively, temperature and salinity showed the lowest variability at the onset of the blooms. Nutrient uptake during the increasing phase of the bloom appeared to affect nitrate concentrations more significantly than those of ammonia and phosphate.     

Spring bloom succession,grazing impact and herbivore selectivity of ciliate communities in response to winter warming

This study aimed at simulating different degrees of winter warming and at assessing its potential effects on ciliate succession and grazing-related patterns. By using indoor mesocosms filled with unfiltered water from Kiel Bight, natural light and four different temperature regimes, phytoplankton spring blooms were induced and the thermal responses of ciliates were quantified. Two distinct ciliate assemblages, a pre-spring and a spring bloom assemblage, could be detected, while their formation was strongly temperature-dependent. Both assemblages were dominated by Strobilidiids; the pre-spring bloom phase was dominated by the small Strobilidiids Lohmaniella oviformis, and the spring bloom was mainly dominated by large Strobilidiids of the genus Strobilidium. The numerical response of ciliates to increasing food concentrations showed a strong acceleration by temperature. Grazing rates of ciliates and copepods were low during the pre-spring bloom period and high during the bloom ranging from 0.06 (Δ0°C) to 0.23 day⁻¹ (Δ4°C) for ciliates and 0.09 (Δ0°C) to 1.62 day⁻¹ (Δ4°C) for copepods. During the spring bloom ciliates and copepods showed a strong dietary overlap characterized by a wide food spectrum consisting mainly of Chrysochromulina sp., diatom chains and large, single-celled diatoms. Priority programme of the German Research Foundation—contribution 4.     

An indoor mesocosm system to study the effect of climate change on the late winter and spring succession of Baltic Sea phyto- and zooplankton

An indoor mesocosm system was set up to study the response of phytoplankton and zooplankton spring succession to winter and spring warming of sea surface temperatures. The experimental temperature regimes consisted of the decadal average of the Kiel Bight, Baltic Sea, and three elevated regimes with 2°C, 4°C, and 6°C temperature difference from that at baseline. While the peak of the phytoplankton spring bloom was accelerated only weakly by increasing temperatures (1.4 days per degree Celsius), the subsequent biomass minimum of phytoplankton was accelerated more strongly (4.25 days per degree Celsius). Phytoplankton size structure showed a pronounced response to warming, with large phytoplankton being more dominant in the cooler mesocosms. The first seasonal ciliate peak was accelerated by 2.1 days per degree Celsius and the second one by 2.0 days per degree Celsius. The over-wintering copepod populations declined faster in the warmer mesocosm, and the appearance of nauplii was strongly accelerated by temperature (9.2 days per degree Celsius). The strong difference between the acceleration of the phytoplankton peak and the acceleration of the nauplii could be one of the “Achilles heels” of pelagic systems subject to climate change, because nauplii are the most starvation-sensitive life cycle stage of copepods and the most important food item of first-feeding fish larvae. Priority programme of the German Research Foundation—contribution 3.     

Pigment signatures reveal temporal and regional differences in taxonomic phytoplankton composition off the west coast of Ireland

The composition of phytoplankton assemblages in April, 1998in Galway Bay and during a summer phytoplankton bloom occurringsouthwest of Ireland in August, 1998, was characterized by pigmentsmeasured by high-performance liquid chromatography. Pigmentdata reflecting phytoplankton assemblages dominated by diatomsin Galway Bay and dinoflagellates in the southwest of Irelandwere compared to phytoplankton cell counts. Significant relationshipswere found between the fucoxanthin concentrations and the diatomcell numbers (P < 0.0002 r² 0.63) during April, and betweenfucoxanthin (P < 0.0001 r² 0.79), 19'hexanoyloxyfucoxanthin(P < 0.0001 r² 0.77) concentrations and Gyrodinium aureolumcell numbers during the summer bloom.     

Dynamics of Baltic Sea phages driven by environmental changes

Phage predation constitutes a major mortality factor for bacteria in aquatic ecosystems, and thus, directly impacts nutrient cycling and microbial community dynamics. Yet, the population dynamics of specific phages across time scales from days to months remain largely unexplored, which limits our understanding of their influence on microbial succession. To investigate temporal changes in diversity and abundance of phages infecting particular host strains, we isolated 121 phage strains that infected three bacterial hosts during a Baltic Sea mesocosm experiment. Genome analysis revealed a novel Flavobacterium phage genus harboring gene sets putatively coding for synthesis of modified nucleotides and glycosylation of bacterial cell surface components. Another novel phage genus revealed a microdiversity of phage species that was largely maintained during the experiment and across mesocosms amended with different nutrients. In contrast to the newly described Flavobacterium phages, phages isolated from a Rheinheimera strain were highly similar to previously isolated genotypes, pointing to genomic consistency in this population. In the mesocosm experiment, the investigated phages were mainly detected after a phytoplankton bloom peak. This concurred with recurrent detection of the phages in the Baltic Proper during summer months, suggesting an influence on the succession of heterotrophic bacteria associated with phytoplankton blooms.     

Response of water column microbial communities to sudden exposure to deltamethrin in aquatic mesocosms

Sudden exposure of an aquatic system to an insecticide can have significant effects on populations other than susceptible organisms. Although this is intuitively obvious, little is actually known about how such exposure might affect bacterial communities and their relative metabolic activity in ecosystems. Here, we assessed small sub-unit (ssu)-RNA levels in open and shaded 9 m(3) aquatic mesocosms (16 units - 2 x 2 factorial design in quadruplicate) to examine the effects of sudden addition of deltamethrin to the units. When deltamethrin was added, a cascade of bacterial then phytoplankton "blooms" occurred over time. The bacterial bloom, which most likely included organisms from the plastid/cyanobacterial phylogenetic guild, was almost immediate (within hours), whereas the phytoplankton (algal) bloom lagged by about 4 days. This sequential response can be explained by an apparent sudden release of nutrients consequent to arthropod death that triggered a series of responses in the microbial loop. Interestingly, bacterial blooms were noted in both open and shaded mesocosms, whereas the algal bloom was only seen in open units, suggesting that both deltamethrin addition (and presumptive nutrient release) and an adequate light supply was required for the phytoplankton response. Overall, this work shows that microbial activities as reflected by ssu-rRNA levels can respond dramatically via apparently indirect effects following insecticide application.     

Stimulation of Alexandrium fundyense growth by bacterial assemblages from the Bay of Fundy

AIMS: To evaluate the influence of marine bacterial assemblages from the Bay of Fundy on Alexandrium fundyense str. CB301 growth. METHODS AND RESULTS: Bacterial assemblages were collected from the Bay of Fundy during an Alexandrium spp. bloom, serially diluted to extinction and inoculated into axenic CB301 cultures. Bacterial assemblages dramatically enhanced CB301 growth. Retrieval and analysis of 16S rDNA fragments revealed an Alteromonas sp. strain to be the only detectable bacterium in all assemblages that promoted A. fundyense and the sole bacterium found in the most dilute inoculum that promoted A. fundyense. While this bacterium has not yet been isolated, other isolates obtained from the assemblages did not stimulate A. fundyense, indicating that the observed stimulation was not a general effect of marine bacteria. CONCLUSIONS: Bay of Fundy marine bacterial assemblages dominated by a member of the family Alteromonadaceae were found to dramatically stimulate growth of A. fundyense. SIGNIFICANCE AND IMPACT OF THE STUDY: These results show that native bacteria have the potential to dramatically promote the growth of A. fundyense and may play an important role in influencing A. fundyense dynamics in the Bay of Fundy.     

Analysis of candidate gene expression patterns of adult male <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> morphotypes in response to a social dominance hierarchy

Because phytoplankton communities exhibit seasonal patterns driven by changes in physical factors, grazing pressure, and nutrient limitations, climate change, in combination with local phosphorus management policies are expected to impact phytoplankton annual dynamic. We used long-term monitoring data from Lake Geneva (from 1974 to 2010) to test if changes in phytoplankton seasonal succession across years is related to re-oligotrophication, inter-annual variability in thermal conditions, and Daphnia sp. density. We used a Bayesian method to identify species assemblages and wavelet analysis to detect transient dynamics in seasonal periodicity. A decrease in phosphorus concentrations appeared to play a major role in the inter-annual replacement of species assemblages. Furthermore, some species assemblages exhibited a change in their seasonal periodicity that was most likely induced by changes in Daphnia sp. density. Finally, we demonstrated that flexibility in the pattern of phytoplankton seasonal successions played a stabilizing role at the community level. The results suggest that phenology and inter-annual changes in seasonal dynamics of phytoplankton assemblages are important components to consider for explaining long-term variability in phytoplankton community.     

Rapid successions affect microbial N-acetyl-glucosamine uptake patterns during a lacustrine spring phytoplankton bloom

The vernal successions of phytoplankton, heterotrophic nanoflagellates (HNF) and viruses in temperate lakes result in alternating dominance of top-down and bottom-up factors on the bacterial community. This may lead to asynchronous blooms of bacteria with different life strategies and affect the channelling of particular components of the dissolved organic matter (DOM) through microbial food webs. We followed the dynamics of several bacterial populations and of other components of the microbial food web throughout the spring phytoplankton bloom period in a pre-alpine lake, and we assessed bacterial uptake patterns of two constituents of the labile DOM pool (N-acetyl-glucosamine [NAG] and leucine). There was a clear genotypic shift within the bacterial assemblage, from fast growing Cytophaga-Flavobacteria (CF) affiliated with Fluviicola and from Betaproteobacteria (BET) of the Limnohabitans cluster to more grazing resistant AcI Actinobacteria (ACT) and to filamentous morphotypes. This was paralleled by successive blooms of viruses and HNF. We also noted the transient rise of other CF (related to Cyclobacteriaceae and Sphingobacteriaceae) that are not detected by fluorescence in situ hybridization with the general CF probe. Both, the average uptake rates of leucine and the fractions of leucine incorporating bacteria were approximately five to sixfold higher than of NAG. However, the composition of the NAG-active community was much more prone to genotypic successions, in particular of bacteria with different life strategies: While 'opportunistically' growing BET and CF dominated NAG uptake in the initial period ruled by bottom-up factors, ACT constituted the major fraction of NAG active cells during the subsequent phase of high predation pressure. This indicates that some ACT could profit from a substrate that might in parts have originated from the grazing of protists on their bacterial competitors.     

Roseobacter and SAR11 dominate microbial glucose uptake in coastal North Sea waters

Bacterial assemblages in coastal pelagic environments are exposed to pronounced temporal and spatial fluctuations in the availability of monomeric substrates. Little is known about the response of particular bacterial groups to such variability. We studied glucose incorporation at various concentrations (0.1-100 nM) by bacteria related to Roseobacter, SAR11, Gammaproteobacteria and Cytophaga-Flavobacteria in coastal North Sea waters in late winter and during a spring phytoplankton bloom dominated by Phaeocystis sp. (March and May 2004 respectively). Both the fraction of glucose-assimilating bacterial cells and the rate of substrate incorporation per active cell were higher in May. The respective contributions of the studied groups to all glucose-assimilating Bacteria were related to substrate concentration. The majority of glucose-incorporating bacterial cells at the lower concentrations were members of the Roseobacter and SAR11 clades. At both time-points the two groups formed approximately equal fractions of all glucose-incorporating bacteria. This was due to a small population of highly active Roseobacter cells and high abundances of SAR11 bacteria with low proportions of glucose-incorporating cells. By contrast, the proportion of active cells from the Cytophaga-Flavobacteria lineage substantially increased at higher levels of available substrate. The determination of concentration-dependent substrate incorporation patterns may help to better understand the different ecophysiological niches of bacterioplankton populations.     

VARIATION IN DCMU-ENHANCED FLUORESCENCE RELATIVE TO CHLOROPHYLL A: CORRELATION WITH THE BROWN TIDE BLOOM1

DCMU–enhanced fluorescence and extracted chlorophyll a were simultaneously measured in Narragansett Bay, Rhode Island and the MERL (Marine Ecosystems Research Laboratory) mesocosms during the 1985 brown tide bloom. Marked differences in the relationship between these variables were observed as the phytoplankton community shifted from dominance by picoalgae to diatoms. The fluorescence to chlorophyll a ratio was significantly (P< 0.05) higher in the mesocosms and the bay when the brown tide species (Aureococcus anophagefferens Hargraves et Sieburth) dominated the phytoplankton community compared with other taxa. Although several factors could have affected the relationship we believe the high ratios are related to the pigment composition and/or small size of the brown tide organism.     

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.     

Response of phytoplankton and bacteria to nutrients and zooplankton: a mesocosm experiment

Although both nutrient inputs and zooplankton grazing are importantto phytoplankton and bacteria in lakes, controversy surroundsthe relative importance of grazing pressure for these two groupsof organisms. For phytoplankton, the controversy revolves aroundwhether zooplankton grazers, especially large cladocerans likeDaphnia, can effectively reduce phytoplankton populations regardlessof nutrient conditions. For bacteria, little is known aboutthe balance between possible direct and indirect effects ofboth nutrients and zooplankton grazing. However, there is evidencethat bacteria may affect phytoplankton responses to nutrientsor zooplankton grazing through direct or apparent competition.We performed a mesocosm experiment to evaluate the relativeimportance of the effects of nutrients and zooplankton grazingfor phytoplankton and bacteria, and to determine whether bacteriamediate phytoplankton responses to these factors. The factorialdesign crossed two zooplankton treatments (unsieved and sieved)with four nutrient treatments (0, 0.5, 1.0 and 2.0 µgphosphorus (P) l^–1 day^–1 together with nitrogen(N) at a N:P ratio of 20:1 by weight). Weekly sieving with 300µm mesh reduced the average size of crustacean zooplanktonin the mesocosms, decreased the numbers and biomass of Daphnia,and increased the biomass of adult copepods. Nutrient enrichmentcaused significant increases in phytoplankton chlorophyll a(4–5x), bacterial abundance and production (1.3x and 1.6x,respectively), Daphnia (3x) and total zooplankton biomass (2x).Although both total phytoplankton chlorophyll a and chlorophylla in the <35 µm size fraction were significantly lowerin unsieved mesocosms than in sieved mesocosms, sieving hadno significant effect on bacterial abundance or production.There was no statistical interaction between nutrient and zooplanktontreatments for total phytoplankton biomass or bacterial abundance,although there were marginally significant interactions forphytoplankton biomass <35 µm and bacterial production.Our results do not support the hypothesis that large cladoceransbecome less effective grazers with enrichment; rather, the differencebetween phytoplankton biomass in sieved versus unsieved zooplanktontreatments increased across the gradient of nutrient additions.Furthermore, there was no evidence that bacteria buffered phytoplanktonresponses to enrichment by either sequestering P or affectingthe growth of zooplankton.     

Succession of sea-ice bacterial communities in the Baltic Sea fast ice

Coastal fast ice and underlying water of the northern Baltic Sea were sampled throughout the entire ice winter from January to late March in 2002 to study the succession of bacterial biomass, secondary production and community structure. Temperature gradient gel electrophoresis (TGGE) and sequencing of TGGE fragments were applied in the community structure analysis. Chlorophyll-a and composition of autotrophic and heterotrophic assemblages were also examined. Overall succession of ice organism assemblages consisted of a low-productive stage, the main algal bloom, and a heterotrophic post-bloom situation, as typical for the study area. The most important groups of organisms in ice in terms of biomass were dinoflagellates, plasticidic flagellates, rotifers and ciliates. Ice bacteria showed a specific succession not directly dependent on the overall succession events of ice organisms. Sequenced 16S rDNA fragments were mainly affiliated to α-, β-, and γ-proteobacterial phyla and Cytophaga–Flavobacterium–Bacteroides-group, and related to sequences from cold environments, also from the Baltic Sea. Temporal clustering of the TGGE fingerprints was stronger than spatial, although lower ice and underlying water communities always clustered together, pointing to the importance of ice maturity and ice–water interactions in shaping the bacterial communities.     

Bacterial community composition in an Arctic phytoplankton mesocosm bloom: the impact of silicate and glucose

In order to study interactions between microorganisms at different nutrient conditions in an arctic environment, a mesocosm experiment was performed in Kongsfjorden, Svalbard (79°N). A phytoplankton bloom was initiated by daily additions of mineral nutrients (ammonium and phosphate) to all mesocosm units. The addition of silicate and glucose, forming a factorial design (+Si/+C, +Si/−C, −Si/+C, −Si/−C), was intended to produce different types of growth rate limitation for the bacterial community. We here focus on the response in bacterial community composition to different nutrient situations. Phytoplankton, bacteria and viruses were enumerated by flow cytometry, while denaturing gradient gel electrophoresis (DGGE) was used to track changes in the bacterial community composition. Our results showed that both glucose and silicate addition affected the bacterial community composition, with the largest effect from glucose. The initial increase in bacterial abundance was most pronounced in the glucose units. After silicate addition, highest bacterial abundance was observed in the silicate treatments where mineral nutrient competition by diatoms was expected to be highest. The major effect of glucose was expressed by the significant separation of the +C and the −C samples at the end of the experiment, while silicate addition resulted in a more stable bacterial community structure. In the unit, given both silicate and glucose, the diatoms were totally outcompeted by the bacterial community. The competitive success of the heterotrophic bacteria in C-replete situations allows the conclusion that the bacteria were not more negatively affected by low temperatures than phytoplankton.