Dynamics of autotrophic picoplankton in Lake Constance

The vertical distribution, biomass concentrations and growthrates of autotrophic picoplankton (APP) were investigated duringthe growing season (March-October) in Lake Constance in differentdepths. Cell numbers determined by epifluorescence microscopyvaried between 1.0 x 10³ and 1.6 times; 10⁵ cells ml^–1depending on season and water depth. Highest concentrationswere recorded above the thermodine in late summer. Numerically,APP consisted almost exclusively of chroococcoid cyanobactena.During lake stratification several peaks of biomass concentrationsoccurred in epilimsietic waters at intervals of 6–8 weeks.In-situ experiments using a dilution technique and dialysisbags revealed that during summer APP population dynamics wereprimarily driven by combined changes of their growth and grazingrates, whereas temperature was less important. Gross growthrates varied between 0.006 and 0.051 h^–1, grazing ratesbetween 0.002 and 0.053 h^–1. On average APP productionwas completely removed by grazing within the microbial community.Ciliates, heterotrophic nanoflagellates and rotifers have beenidentified as the major consumers of APP cells. APP biomassis small compared to larger phytoplankton, ranging from ito5% of total phytoplankton biovolume. Due to its high gross growthrates, which are on the same level as those of free-living pelagicbacteria, APP contributes slightly more to overall primary productionwith maximum percentages of {small tilde}15% in late summer.     

The water bloom of Cyanobacterial picoplankton in Lake Biwa,Japan

Unicellular autofluorescent picoplankton ranging from 0.4 to 1.5 µm in diameter were found to be a significant component of phytoplankton in the North Basin of Lake Biwa during early summer in 1989 and 1990. The abundance of these picoplankton varied seasonally by about three orders of magnitude with one maximum of up to 10⁶ cells ml^–1. Bloom-forming picoplankton were isolated by dilution and further cultivated in liquid medium. Three clones were found to be representative species of the bloom. Using epifluorescence and electron microscopy as well as absorption and fluorescence emission spectroscopy, we examined these clones according to shape and pigment composition. They have ringlike thylakoids, are photosynthetically active and have no nuclear envelope. The cyanobacterial clones isolated represent three types containing phycobilisomes with either phycocyanin or phycoerthrin as the dominant accessory pigment. They are described here as three new species, two phycoerythrin-rich types and one phycocyanin-rich type, all of them belonging to the Synechococcus group. The differences found by fluorescence emission of isolated clones are discussed with respect to in situ strain identification.     

Persistence and genetic diversity among strains of phycoerythrinrich cyanobacteria from the picoplankton of Lake Constance

Twelve strains of phycoerythrin (PE)-rich unicellular cyanobacteriaof the Synechococcus type were isolated from the pelagial ofLake Constance in different phases during the growth periodof the year 1994. By analysing the restriction fragment lengthpolymorphism (RFLP) of the DNA three new genotypes were distinguished.The persistence of one strain during the course of the yearwas demonstrated. The data set was compared with the RFLP observedin PE-rich Synechococcus strains isolated in former years fromthe same sampling site. The plurality in the picoplanktic isolatesand the possibility of cultivation of strains is discussed withrespect to the oligotrophication of Lake Constance.     

Growth characteristics of small and large free-living and attached bacteria in Lake Constance

The growth characteristics of small (0.2–1.0m) and large (1.0–3.0 (m) free-living and attached bacteria were studied in Lake Constance by comparing the spatial and seasonal dynamics of their biomass turnover time (ratio of biomass/production). The biomass of small free-living bacteria usually turned over significantly faster than that of large free-living bacteria throughout the water column. The turnover of attached bacterial biomass was characterized by large fluctuations. Occasionally, in aphotic water layers, it was as long as that of large free-living bacteria, but when large amounts of decaying organic particles were present, it was shorter than that of small free-living cells. Biomass turnover times of free-living bacteria were in the same range as their generation times, which were estimated from the increase in bacterial abundance in 3m prefiltered samples. The biomass turnover time of actively metabolizing bacteria was comparable to the generation time of actively metabolizing cells. These results indicate that the biomass turnover time is a useful indicator of the growth of different bacterial fractions, as it reflects their different amounts of participation in microbial processes of aquatic ecosystems.     

Fate of methane bubbles released by pockmarks in Lake Constance

In the eastern part of Lake Constance, the second largest pre-alpine lake in Europe, about five hundred pockmarks (morphological depressions on the lake floor) were recently discovered of which ~40 % release methane bubbles. The carbon isotopic composition of the escaping gas indicated that the methane is of biogenic origin. In our study, we investigated the fate of the released methane bubbles, i.e., the dissolution, oxidation or transport of the bubbles to the surface. At a littoral pockmark site (PM12, 12 m water depth) and a profundal pockmark (PM80, 80 m water depth), we analysed the dissolved methane concentrations and the methane isotopic carbon signature in the water column. At PM80, higher methane concentrations (up to 1,523 nM), compared to the control site and the surface waters (225 ± 72 nM), were recorded only on some occasions and only in the bottom water, despite the fact that the released bubbles were dissolving within the hypolimnion based on bubble modeling. The isotope data suggest that most of the dissolved methane is oxidized below 40 m water depth. The isotopic signature of the methane in the surface water at PM80, however, differed from that of the methane in the hypolimnion; therefore, the surface methane at this profundal site is most likely an export product from the littoral zone. Assuming an initial bubble diameter of 5 mm, we calculated that these small bubbles would reach the surface, but approximately 96 % of the methane would have dissolved from the bubble into the hypolimnion. At PM12, we observed higher concentrations of dissolved methane (312 ± 52 nM) with no significant differences between seasons or between control sites versus pockmark site. In the shallow water, divers estimated the bubble size to be 10–15 mm, which from a release depth of 12 m would barely dissolved into the water column. The isotopic signature also indicated that there had been almost no methane oxidation in the shallow water column. Thus, the water depth of bubble release as well as the initial bubble size determine whether the methane enters the atmosphere largely unhindered (shallow site) or if the released methane is incorporated into the profundal water column.     

Microcystins in Cyanobacterial Biofilms from the Littoral Zone of Lake Baikal

Some species of cyanobacteria synthesize toxins whose concentration during water bloom can reach values dangerous for human and animal health. Planktonic cyanobacteria are the most common and well-studied microcystins producers, hepatotoxic cyclic heptapeptides, whereas microcystin-producing benthic cyanobacteria are less known. In recent years, the mass development of benthic cyanobacteria forming extensive fouling on different substrates has been detected in the littoral zone of Lake Baikal. We found microcystins produced by benthic cyanobacteria in the biofouling on different natural and artificial substrates, including diseased and dead endemic sponges Lubomirskia baicalensis and Baikalospongia spp. collected from the littoral area of Lake Baikal. Microscopic analysis of the biofouling revealed prevalence of representatives of Nostocales and Oscillatoriales with predominance of Tolypothrix distorta that is likely the main microcystin producer in Lake Baikal. According to enzyme-linked immunosorbent assay (ELISA), microcystin concentrations in biofouling were 29.8–3050 μg/kg dry weight. We identified eight microcystin variants using MALDI-TOF/TOF; [Dha7]MC-YR was detected in most samples. The presence of microcystins in biofilms formed on the surface of the artificial substrate by Phormidium autumnale was also recorded. The data obtained demonstrated the necessity to monitor potentially toxic species and concentrations of cyanotoxins in plankton and benthos in the littoral zone of Lake Baikal, especially in the regions with intense tourist and recreational activities.     

Biosorption of Copper by Cyanobacterial Bloom-Derived Biomass Harvested from the Eutrophic Lake Dianchi in China

Biomass of cyanobacterial bloom from Lake Dianchi was used as a biosorbent for copper removal from aqueous solution. The maximum capacity was found at conditions of pH 4, initial concentration of copper was 10 mg/l and initial dose of biomass was 1.0 g/l. HNO₃ demonstrated the highest desorption efficiency compared with HCl, EDTA, and citric acid. Physical adsorption was assumed not to be the dominant mechanism of biosorption as revealed by scanning electron microscopy and surface area measurement of the biomass. Infrared ray spectra analysis of the biomass suggested that ion-exchange is the principal mechanism for biosorption. Considering the advantages—low cost, easy to collect, and huge in quantity—the Microcystis bloom biomass could be used as a sorbent for copper and other heavy metals removal.     

Genetic diversity among Synechococcus spp. (cyanobacteria) isolated from the pelagial of Lake Constance

Abstract: Seven phycoerythrin (PE)-rich and six phycocyanin (PC)-rich unicellular cyanobacteria of the Synechococcus type were isolated from the pelagial of Lake Constance. The genetic diversity among the isolates was evaluated using restriction fragment length polymorphism (RFLP) within the psbA gene family. Nine out of 13 isolates exhibit different DNA structures in the probed areas and, furthermore, they differ from morphologically similar strains collected from other lakes. The data set does not support a principal distinction between PC-rich and PE-rich strains but it reveals less heterogeneity in the coding region of the psbA genes among PE-rich isolates than among PC-rich isolates. The isolation of distinct strains in different seasons suggests species diversity and seasonal occurrence of Synechococcus spp. in Lake Constance.     

Isolation of novel pelagic bacteria from the German bight and their seasonal contributions to surface picoplankton.

We tested new strategies for the isolation of abundant bacteria from coastal North Sea surface waters, which included reducing by several orders of magnitude the concentrations of inorganic N and P compounds in a synthetic seawater medium. Agar plates were resampled over 37 days, and slowly growing colonies were allowed to develop by repeatedly removing all newly formed colonies. A fivefold increase of colonies was observed on plates with reduced nutrient levels, and the phylogenetic composition of the culture collection changed over time, towards members of the Roseobacter lineage and other alpha-proteobacteria. Novel gamma-proteobacteria from a previously uncultured but cosmopolitan lineage (NOR5) formed colonies only after 12 days of plate incubation. A time series of German Bight surface waters (January to December 1998) was screened by fluorescence in situ hybridization (FISH) with isolate-specific and general probes. During spring and early summer, a prominent fraction of FISH-detectable bacteria (mean, 51%) were affiliated with the Cytophaga-Flavobacterium group (CF) of the Bacteroidetes. One Cytophaga sp. lineage with cultured representatives formed almost 20% of the CF group. Members of the Roseobacter cluster constituted approximately 50% of alpha-proteobacteria, but none of the Roseobacter-related isolates formed populations of >1% in the environment. Thus, the readily culturable members of this clade are probably not representative of Roseobacter species that are common in the water column. In contrast, members of NOR5 were found at high abundances (>10(5) cells ml(-1)) in the summer plankton. Some abundant pelagic bacteria are apparently able to form colonies on solid media, but appropriate isolation techniques for different species need to be developed.     

Diel Nitrogen Fixation by Cyanobacterial Surface Blooms in Sanctuary Lake, Pennsylvania

Diel nitrogen fixation studies were conducted with assemblages of cyanobacteria sampled from surface blooms on Sanctuary Lake, Pa. The studies were conducted between July and September of 1982 to 1985 by using the acetylene reduction technique. Assemblages with the lowest cell concentrations (0.9 × 10⁹ to 1.0 × 10⁹ cells per liter) exhibited nitrogen fixation activity throughout the day, with maximum fixation rates occurring in mid to late afternoon; fixation proceeded throughout the night at rates equivalent to 23 to 28% of the afternoon maximum. In studies conducted with the highest cell concentrations (3.7 × 10⁹ to 6.7 × 10⁹ cells per liter), fixation rates reached maximum values in mid to late morning. The rates declined rapidly throughout the midday period and subsequently ceased from late afternoon until sunrise on the following day. The afternoon decline and cessation of fixation exhibited by high cell concentrations correlated with photosynthetically induced low total CO₂ and supersaturating O₂ concentrations. The midday decline could be prevented and partially reversed by experimentally lowering O₂ and increasing total CO₂ concentrations. Under experimental conditions which simultaneously prevented supersaturating O₂ concentrations and maintained high total CO₂ availability, nitrogen fixation continued throughout the solar day, with maximum rates occurring at midday. These observations indicate that temporal changes in photosynthetic activity may affect diel fluctuations in nitrogen fixation.     

Morphological and phylogenetic characterizations of freshwater Thioploca species from Lake Biwa,Japan, and Lake Constance,Germany

Filamentous, gliding, sulfide-oxidizing bacteria of the genus Thioploca were found on sediments in profundal areas of Lake Biwa, a Japanese freshwater mesotrophic lake, and were characterized morphologically and phylogenetically. The Lake Biwa Thioploca resembled morphologically Thioploca ingrica, a brackish water species from a Danish fjord. The diameters of individual trichomes were 3 to 5.6 microm; the diameters of complete Thioploca filaments ranged from 18 to 75 micro m. The cell lengths ranged from 1.2 to 3.8 micro m. In transmission electron microscope specimens stained with uranyl acetate, dense intracellular particles were found, which did not show any positive signals for phosphorus and sulfur in an X-ray analysis. The 16S rRNA gene of the Thioploca from Lake Biwa was amplified by using newly designed Thioploca-specific primers (706-Thioploca, Biwa160F, and Biwa829R) in combination with general bacterial primers in order to avoid nonspecific amplification of contaminating bacterial DNA. Denaturing gradient gel electrophoresis (DGGE) analysis of the three overlapping PCR products resulted in single DGGE bands, indicating that a single 16S rRNA gene had been amplified. With the same method, the Thioploca from Lake Constance was examined. The 16S rRNA sequence was verified by performing fluorescence in situ hybridization targeted at specific motifs of the Lake Biwa THIOPLOCA: Positive signals were obtained with the bacterial probe EUB-338, the gamma-proteobacterial probe GAM42a, and probe Biwa829 targeting the Lake Biwa THIOPLOCA: Based on the nearly complete 16S rRNA sequence and on morphological similarities, the Thioploca from Lake Biwa and the Thioploca from Lake Constance are closely related to T. ingrica and to each other.     

Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy.

Accurate measurement of the biomass and size distribution of picoplankton cells (0.2 to 2.0 microns) is paramount in characterizing their contribution to the oceanic food web and global biogeochemical cycling. Image-analyzed fluorescence microscopy, usually based on video camera technology, allows detailed measurements of individual cells to be taken. The application of an imaging system employing a cooled, slow-scan charge-coupled device (CCD) camera to automated counting and sizing of individual picoplankton cells from natural marine samples is described. A slow-scan CCD-based camera was compared to a video camera and was superior for detecting and sizing very small, dim particles such as fluorochrome-stained bacteria. Several edge detection methods for accurately measuring picoplankton cells were evaluated. Standard fluorescent microspheres and a Sargasso Sea surface water picoplankton population were used in the evaluation. Global thresholding was inappropriate for these samples. Methods used previously in image analysis of nanoplankton cells (2 to 20 microns) also did not work well with the smaller picoplankton cells. A method combining an edge detector and an adaptive edge strength operator worked best for rapidly generating accurate cell sizes. A complete sample analysis of more than 1,000 cells averages about 50 min and yields size, shape, and fluorescence data for each cell. With this system, the entire size range of picoplankton can be counted and measured.     

Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy.

Accurate measurement of the biomass and size distribution of picoplankton cells (0.2 to 2.0 microns) is paramount in characterizing their contribution to the oceanic food web and global biogeochemical cycling. Image-analyzed fluorescence microscopy, usually based on video camera technology, allows detailed measurements of individual cells to be taken. The application of an imaging system employing a cooled, slow-scan charge-coupled device (CCD) camera to automated counting and sizing of individual picoplankton cells from natural marine samples is described. A slow-scan CCD-based camera was compared to a video camera and was superior for detecting and sizing very small, dim particles such as fluorochrome-stained bacteria. Several edge detection methods for accurately measuring picoplankton cells were evaluated. Standard fluorescent microspheres and a Sargasso Sea surface water picoplankton population were used in the evaluation. Global thresholding was inappropriate for these samples. Methods used previously in image analysis of nanoplankton cells (2 to 20 microns) also did not work well with the smaller picoplankton cells. A method combining an edge detector and an adaptive edge strength operator worked best for rapidly generating accurate cell sizes. A complete sample analysis of more than 1,000 cells averages about 50 min and yields size, shape, and fluorescence data for each cell. With this system, the entire size range of picoplankton can be counted and measured.