Andreas Brutemark

Abstract: Bacterioplankton growth is often directly or indirectly controlled by external energy subsidies via organic matter inputs or solar radiation. We carried out a mesocosm experiment to assess how bacterioplankton communities responded to elevated levels of dissolved organic matter (DOM) and experimentally controlled stratification depth. The month-long experiment consisted of 2500 l mesocosms subjected to 4 experimental manipulations in triplicate: the stratification depth was set to either 1.5 m or 3.5 m with or without experimental addition of ambient levels of chromophoric DOM. DOM addition had a significant effect on bacterial community composition assessed by terminal restriction fragment length polymorphism of amplified 16S rRNA genes. In contrast, there were no effects of the DOM amendment on bacterial biomass or production. Mixing depth and the coupled effective light climate in the photic zone also had a significant effect on bacterial community composition. Shallow mixing depth was also associated with enhanced primary production whereas the DOM addition had a negative effect on phytoplankton biomass and productivity. Our results suggest that bacterial community composition is coupled to primary production under the studied coastal nutrient regime, and point to a key role of DOM quality in controlling bacterioplankton communities.

Abstract: Bioassay experiments were performed two times (with 2 years in between) in order to investigate if nitrogen (N, ammonium), phosphorus (P, phosphate) and carbon (C, glucose) additions would stimulate the growth of bacteria and phytoplankton differently in three different tropical aquatic environments. The water and their indigenous microbial communities were taken from a freshwater coastal lake (Cabiunas), a coastal (Anjos), and an offshore marine station (Sonar) in the Atlantic outside Cabo Frio, Rio de Janeiro State, Brazil. Ammonium, phosphate and glucose were added alone or in combination to triplicate bottles. In the lake, P seemed to be the primary limiting factor during the first experiment, since both bacterial production and phytoplankton growth was stimulated by the P addition. Two years later, however, addition of P inhibited phytoplankton growth. During both years, C was closely co-limiting for bacteria since CP additions increased the response considerably. For both the coastal and offshore seawater stations, phytoplankton growth was clearly stimulated by N addition in both years and the bacteria responded either to the P, N or C additions (alone or in combination). To conclude, the results from these tropical aquatic systems show that it is possible that phytoplankton and bacteria may compete for a common resource (P) in lakes, but can be limited by different inorganic nutrients in marine waters as well as lakes, suggesting that phytoplankton and bacteria do not necessarily compete for the same growth limiting nutrient in these environments.

Abstract: Monitoring data on water pH are presented for the period between 1972 and 2009 from the sampling stations Längden and Storgadden, at the entrance to the Gulf of Finland, Baltic Sea. The overall pH in the area ranged from 9.2 to 7.4, on average 8.1, and showed a significant decreasing trend during the winter period corresponding to a median annual decrease of 0.006. The data corroborate previous findings about a seasonal effect, where pH is higher during summer than winter.

Abstract: Mixotrophy in Prymnesium parvum was investigated using carbon (d13C) and nitrogen (d15N) stable isotopes. The experiment was performed in light and dark. In the dark treatment we expected that the mixotrophic P. parvum would rely solely on its prey and therefore reflect the prey isotopic signatures. In the light treatment P. parvum can perform photosynthesis as well as utilize its prey, thus we expect the isotopic signatures to be between the dark mixed cultures and the monocultures, depending on how much prey was utilized. In the light treatment, the addition of the ciliate Myrionecta rubra resulted in higher P. parvum cell numbers compared to monocultures. During the experiment, cell numbers in the dark monocultures and the mixed dark cultures did not increase. P. parvum had 2.5�3 times higher cellular phosphorus and nitrogen content in the dark compared to the cultures in the light whereas no difference in carbon content between treatments could be observed. This suggests that P. parvum can utilize nitrogen and phosphorus but not carbon in the dark. It thus seems as if P. parvum relies on photosynthesis to meet the carbon and energy demand required for growth. The expected isotopic signatures ��become what you eat. . . plus a few per mil�� were not observed. In the dark treatment, the d13C did not differ between monocultures and mixed cultures. In the light treatments P. parvum d13C became less negative then the corresponding dark treatments indicating that P. parvum used CO2 rather than carbon from the added prey. No difference in d15N betweenmonocultures and mixed cultures could be observed during the experiment. We argue that light is necessary for P. parvum growth and that the ability to utilize nutrients originating from their prey may be important in bloom formation.

Abstract: Harmful algal bloom species can persist in the environment, impacting aquatic life and human health. One of the mechanisms by which some harmful algal bloom species are able to persist is by consumption of organic particles. Methods to demonstrate and measure consumption can yield insight into how populations thrive. Here, we combine flow cytometry and real-time PCR to demonstrate consumption of a cryptophyte species (Rhodomonas sp.) by a toxic mixotrophic haptophyte (Prymnesium parvum). Using flow cytometry, the feeding frequency of a population of P. parvum cells was calculated using the phycoerythrin (PE) fluorescence signal from Rhodomonas sp. and the fluorescence of an acidotropic probe labeling the food vacuoles. Feeding frequency increased in the beginning of the experiment and then began to decline, reaching a maximum of 47.5% of the whole P. parvum population after 212 min. The maximum number of consumed Rhodomonas sp. cells was 0.8 per P. parvum cell, and occurred after 114 min corresponding to an ingestion rate of 0.4 Rhodomonas sp. cells/P. parvum/h. Cells from the feeding P. parvum population were sorted, washed, and subjected to a real-time PCR assay targeting the cryptophyte 18S locus. There was a correlation between cycle threshold (Ct) values and number of consumed prey cells calculated by fluorescence. Overall, this study shows that flow cytometric analysis, of the acidotropic probe and prey pigments, is an efficient and rapid tool in enumerating food vacuoles and the number of prey cells consumed. Furthermore, we suggest that real-time PCR can be applied to cells sorted by flow cytometry, thus allowing for the detection and potential quantification of the targeted prey cells.

Abstract: In this study we have investigated whether the carbon isotopic signature differs between different groups and species of marine phytoplankton depending on growth phase, nutrient conditions and salinity. The 15 investigated algal species, representing the Bacillariophyceae, Chlorophyceae, Cryptophyceae, Cyanophyceae, Dinophyceae and Haptophyceae classes were grown in batch monocultures and analysed for δ13C in both exponential and stationary phase. For all the cultured species, δ13C signatures ranged from � 23.5� (Imantonia sp.) to � 12.3� (Nodularia spumigena) in the exponential phase and from � 18.8� (Amphidinium carterae) to � 8.0� (Anabaena lemmermannii) in the stationary phase. Three species (Dunaliella tertiolecta, Rhodomonas sp., Heterocapsa triquetra) were also grown under nutrient sufficient and nitrogen or phosphorus deficient conditions. Nitrogen limitation resulted in a more negative δ13C signature, whereas no effect could be observed during phosphorus limitation compared to nutrient sufficient conditions. Growth of Prymnesium parvum in two different salinities resulted in a more negative δ13C signature in the 26�-media compared to growth in 7�-media. Our results show that the carbon isotopic signature of phytoplankton may be affected by salinity, differ among different phytoplankton species, between exponential and stationary phase, as well as between nutrient treatments.

Abstract: We investigated whether phytoplankton communities in two lakes in SW Greenland were phosphorus or nitrogen limited. The study lakes have contrasting water chemistry (mean conductivities differ ten fold) and are located near Kangerlussuaq, SW Greenland (~67°N, 51°W). A microcosm nutrient enrichment experiment was performed in June 2003 to determine whether nitrate or phosphate addition stimulated phytoplankton growth. Samples were analysed for species composition, biomass, and alkaline phosphatase activity (APA). Initially, both lakes had extremely low total phosphorus but high total nitrogen concentrations and high APA, suggesting that the phytoplankton were phosphorus limited prior to the start of the experiment. The phytoplankton composition and biomass (mainly Ochromonas spp.) responded to phosphate but not to nitrate addition. In both lakes, chlorophyll a increased significantly when phosphate was added. Furthermore, APA was significantly lower in the two lakes when phosphate was added compared to the control and the nitrogen addition treatment. The dominance of mixotrophic phytoplankton and high DOC values suggest that these lakes may be regulated by microbial loop processes.