蓝藻胞外多聚物生物合成、群体形成与微囊藻水华

有毒微囊藻水华在太湖、巢湖和滇池等饮用水源地频繁暴发,对居民健康和水产养殖等构成严重威胁,亟需开发新技术加以有效控制和利用。在水华暴发时,蓝藻大量分泌胞外多聚物而形成细胞群体,是蓝藻水华发生的关键和前提。蓝藻群体中胶质状胞外多聚物由胞外多糖、蛋白质和其他生物大分子组成,对其结构、功能和生物合成途径研究了解仍然有限。生物信息学和比较基因组学分析发现微囊藻和其他多种蓝藻中编码大量的具有称之为PEP-CTERM结构域的潜在胞外蛋白质,这些潜在的蛋白质可能通过特殊的分选系统分泌到细胞表面,与胞外多糖相互作用形成结构更复杂的胞外多聚物,介导细胞群体的形成和水华发生。亟需建立微囊藻遗传操作技术,深入揭示胞外多聚物生物合成和群体形成的分子机制,寻找控制蓝藻胞外多聚物的组装和分泌及群体形成的关键靶点,将有助于揭示蓝藻水华形成机理及开发新型控藻技术。     

Size-dependent growth rates in eukaryotic and prokaryotic algae exemplified by green algae and cyanobacteria: comparisons between unicells and colonial growth forms

The size dependency of maximum growth rates was investigatedin cyanobacteria and in green algae (Chlorophyta). Both unicellularand colony-forming species were included in the study. Significantallometric relationships were found between size and maximumgrowth rate for both cyanobacteria and green algae. The size-dependentgrowth could be described by the same scaling exponent in bothcyanobacteria and green algae, but in both cyanobacteria andgreen algae only unicells evinced size-dependent growth rates—therewas no relationship between colony size and growth rate in colonialforms of cyanobacteria and green algae. It is concluded thatthe colonial growth form represents an evolutionary adaptationto escape the negative effects of size-dependent growth, whileretaining the positive effects of increased size, e.g. a decreasedgrazing pressure.     

Measuring carbon and N2 fixation in field populations of colonial and free‐living unicellular cyanobacteria using nanometer‐scale secondary ion mass spectrometry1

Unicellular cyanobacteria are now recognized as important to the marine N and C cycles in open ocean gyres, yet there are few direct in situ measurements of their activities. Using a high‐resolution nanometer scale secondary ion mass spectrometer (nanoSIMS), single cell N₂ and C fixation rates were estimated for unicellular cyanobacteria resembling N₂ fixer Crocosphaera watsonii. Crocosphaera watsonii‐like cells were observed in the subtropical North Pacific gyre (22°45′ N, 158°0′ W) as 2 different phenotypes: colonial and free‐living. Colonies containing 3–242 cells per colony were observed and cell density in colonies increased with incubation time. Estimated C fixation rates were similarly high in both phenotypes and unexpectedly for unicellular cyanobacteria 85% of the colonial cells incubated during midday were also enriched in ¹⁵N above natural abundance. Highest ¹⁵N enrichment and N₂ fixation rates were found in cells incubated overnight where up to 64% of the total daily fixed N in the upper surface waters was attributed to both phenotypes. The colonial cells retained newly fixed C in a sulfur‐rich matrix surrounding the cells and often cells of both phenotypes possessed areas (<1 nm) of enriched ¹⁵N and ¹³C resembling storage granules. The nanoSIMS imaging of the colonial cells also showed evidence for a division of N₂ and C fixation activity across the colony where few individual cells (<34%) in a given colony were enriched in both ¹⁵N and ¹³C above the colony average. Our results provide new insights into the ecophysiology of unicellular cyanobacteria.     

Seasonal Dynamics of Picocyanobacteria and Picoeukaryotes in a Large Shallow Lake (Lake Balaton,Hungary)

The abundance and composition of autotrophic picoplankton (APP) were studied between February 2003 and March 2004 in Lake Balaton. Water samples were taken fortnightly in the eutrophic western basin and mesotrophic eastern basin. Our study, which took more than one year, revealed pronounced seasonal pattern of the picoplankton abundance and composition. According to our results there were three types of picoplankton in Lake Balaton: 1. Phycoerythrin‐rich coccoid cyanobacteria (PE), dominant summer picoplankters in the mesotrophic lake area; 2. Phycocyanin‐rich cyanobacteria (PC), the most abundant summer picoplankters in the eutrophic lake area; 3. Picoeukaryotes, dominant winter picoplankters in the whole lake. The observed abundance of picoeukaryotes (3 × 10⁵ cells ml^–1) was one of the highest ever found. Our study confirms that in Lake Balaton the colonial autotrophic picoplankton (colonial APP) become dominant in summer in the nutrient limited period. We have found strong negative relationship between the concentrations of available nitrogen forms (NH₄–N, NO₃–N, urea‐N) and the colonial APP abundance. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Zooplankton grazing on bacteria and cyanobacteria in a eutrophic lake

During in situ experiments in a cyanobacteria- and copepod-dominatedeutrophic subtropical lake, all taxa of protozoa and metazoanzooplankton grazed fluorescently labelled bacteria and all metazoanzooplankton taxa grazed large filamentous and colonial cyanobacteria.     

SHIFT FROM CHLOROPHYTES TO CYANOBACTERIA IN BENTHIC MACROALGAE ALONG A GRADIENT OF NITRATE DEPLETION1

A survey of the spatial distribution of benthic macroalgae in a fluvial lake of the St. Lawrence River (Lake Saint‐Pierre, Quebec, Canada) revealed a shift in composition from chlorophytes to cyanobacteria along the flow path of nutrient‐rich waters originating from tributaries draining farmlands. The link between this shift and changes in water quality characteristics was investigated by sampling at 10 sites along a 15 km transect. Conductivity, current, light extinction, total phosphorus (TP; >25 μg P · L^?1), and ammonium (8–21 μg N · L^?1) remained fairly constant along the transect in contrast to nitrate concentrations, which fell sharply. Filamentous and colonial chlorophytes [Cladophora sp. and Hydrodictyon reticulatum (L.) Bory] dominated in the first 5 km where nitrate concentrations were >240 μg N · L^?1. A mixed assemblage of chlorophytes and cyanobacteria characterized a 1 km transition zone where nitrate decreased to 40–80 μg N · L^?1. In the last section of the transect, nitrate concentrations dropped below 10 μg N · L^?1, and cyanobacteria (benthic filamentous mats of Lyngbya wollei Farl. ex Gomont and epiphytic colonies of Gloeotrichia) dominated the benthic community. The predominance of nitrogen‐fixing, potentially toxic cyanobacteria likely resulted from excessive nutrient loads and may affect nutrient and trophic dynamics in the river.     

Algal and Cyanobacterial Biofilms on Calcareous Historic Buildings

Major microorganisms in biofilms on external surfaces of historic buildings are algae, cyanobacteria, bacteria, and fungi. Their growth causes discoloration and degradation. We compared the phototrophs on cement-based renderings and limestone substrates at 14 historic locations (47 sites sampled) in Europe and Latin America. Most biofilms contained both cyanobacteria and algae. Single-celled and colonial cyanobacteria frequently constituted the major phototroph biomass on limestone monuments (32 sites sampled). Greater numbers of phototrophs, and especially of algae and of filamentous morphotypes, were found on cement-based renderings (15 sites), probably owing to the porosity and small pore size of the latter substrates, allowing greater entry and retention of water. All phototrophic groups were more frequent on Latin American than on European buildings (20 and 27 sites, respectively), with cyanobacteria and filamentous phototrophs showing the greatest differences. The results confirm the influence of both climate and substrate on phototroph colonization of historic buildings. Received: 7 March 2002 / Accepted: 8 April 2002     

Cyanobacteria as indicators of organic contamination

In two Costa Rican rivers used as receptors for domestic sewage, treated by primary stabilization ponds, were taken a total of 28 samplings located at the pond exit and at three different sites in each river: 100 m before the ponds discharge, at the discharge and 100 m after the discharge. These sampling were done for a five and a half years including dry and rainy seasons. In each sampling site, samples were collected of five different substrates: stones, submerge and semi submerge vegetation, tree trunks or sticks, water and artificial substrates. For each sample were used two types of artificial cultures, WC and BG110. A total of 55 cyanobacteria species isolations were obtained, belonging to a 26 genera, between these the most common were Phormidium with nine species, Microcystis with five species, Leptolyngbya and Pseudanabaena with four species each and Oscillatoria with three species. More cyanobacteria species were isolated in water substrate and less isolations in tree trunks and submerge vegetation. Konvophoron, Cyanarcus and Pilgeria only were isolate from water samples inoculated in culture media WC and in few opportunities, while three Leptolyngbya species and four Phormidium species were isolated very often. At the stabilization ponds Phormidium sp4 was dominant in 25 of 28 sampling while in the last others were the chlorophycea I. In this study were observed an increase in the frequency of cyanobacteria at the higher contamination places, and a species substitution between different sampling points. There were no biomass studies, therefore is not possible to relate between different cyanobacteria species and some specific types of water quality.     

La plasticité du genre Kobyastraea (Hexacorallia): Un bon marqueur paléoécologique

New forms of the genus KobyastraeaRoniewiczhave been found in the Bajocian from Jura and Lorraine (France). They are adapted to low light levels (colonial morphology, colonial structure, calicinal structure).     

Do fish regulate phytoplankton in shallow eutrophic Northeast Brazilian reservoirs?

SUMMARY 1. In a comparative study, we examined the potential for fish to structure planktonic food webs in shallow mesotrophic to hypereutrophic Northeast Brazilian reservoirs. The food webs were dominated by three guilds of fish (facultative piscivores, generalist planktivores and omnivores), small herbivorous zooplankton and bloom‐forming cyanobacteria, with few littoral macrophytes. 2. A principal component's analysis on data from 13 reservoirs (27 sampling dates in 1995–99) revealed that euphotic depth, the relative density of phytoplankton (i.e. the percentage of overall phytoplankton density) represented by cyanobacteria, and the relative biomass of fish (i.e. percentage of overall biomass) represented by omnivores and facultative piscivores, explained most of the variance in the data. Physico‐chemical conditions, lake morphometry and rainfall were secondary factors. 3. Phytoplankton was related to fish guild structure. Chlorophyll concentration increased with total phosphorus and the relative biomass of omnivorous fish, decreased with the relative biomass of facultative piscivores, but was unrelated to the biomass and mean body size of herbivorous zooplankton. Chlorophyll concentration and the densities of filamentous and colonial cyanobacteria decreased with the ratio of the biomass of facultative piscivores to that of omnivores (FP : OM). 4. We propose two complementary mechanisms for the observed relationships between fish and phytoplankton. At a low biomass of facultative piscivores, juvenile zooplanktivorous fishes may induce a trophic cascade on zooplankton in the littoral zone. Regardless of piscivore biomass, piscivores and omnivores may regulate phytoplankton via multichannel omnivory because of the predominance of omnivorous or detritivorous foraging behaviour. 5. Manipulative experiments are needed to explore further whether, depending on priorities in the use of the reservoir, fisheries management could alter the FP : OM ratio either to enhance fish yields or to reduce phytoplankton densities and cyanobacterial blooms.     

Grazing on colonial and filamentous, toxic and non-toxic cyanobacteria by the zebra mussel Dreissena polymorpha

Colony forming and toxic cyanobacteria form a problem in surfacewaters of shallow lakes, both for recreation and wildlife. Zebramussels, Dreissena polymorpha, have been employed to help torestore shallow lakes in the Netherlands, dominated by cyanobacteria,to their former clear state. Zebra mussels have been presentin these lakes since they were created in the 19th century bythe excavation of peat and are usually not considered to bean invasive species. Most grazing experiments using Dreissenahave been performed with uni-cellular phytoplankton laboratorystrains and information on grazing of larger phytoplankton taxahardly exists. To gain more insight in to whether D. polymorphais indeed able to decrease cyanobacteria in the phytoplankton,we therefore performed grazing experiments with zebra musselsand two species of cyanobacteria, that greatly differ in shape:colony forming strains of Microcystis aeruginosa and the filamentousspecies Planktothrix agardhii. For both species a toxic anda non-toxic strain was selected. We found that zebra musselscleared toxic Planktothrix at a higher rate than non-toxic Planktothrix,toxic or non-toxic Microcystis. Clearance rates between theother strains were not significantly different. Both phytoplanktonspecies, regardless of toxicity, size and shape, were foundin equal amounts (based on chlorophyll concentrations) in theexcreted products of the mussels (pseudofaeces). The resultsshow that zebra mussels are capable of removing colonial andfilamentous cyanobacteria from the water, regardless of whetherthe cyanobacteria are toxic or not. This implies that the musselsmay be used as a biofilter for the removal of harmful cyanobacterialblooms in shallow (Dutch) lakes where the mussels are alreadypresent and not a nuisance. Providing more suitable substratefor zebra mussel attachment may lead to appropriate mussel densitiescapable of filtering large quantities of cyanobacteria.     

Algal Diet of Small-Bodied Crustacean Zooplankton in a Cyanobacteria-Dominated Eutrophic Lake

Small-bodied cladocerans and cyclopoid copepods are becoming increasingly dominant over large crustacean zooplankton in eutrophic waters where they often coexist with cyanobacterial blooms. However, relatively little is known about their algal diet preferences. We studied grazing selectivity of small crustaceans (the cyclopoid copepods Mesocyclops leuckarti, Thermocyclops oithonoides, Cyclops kolensis, and the cladocerans Daphnia cucullata, Chydorus sphaericus, Bosmina spp.) by liquid chromatographic analyses of phytoplankton marker pigments in the shallow, highly eutrophic Lake Võrtsjärv (Estonia) during a seasonal cycle. Copepods (mainly C. kolensis) preferably consumed cryptophytes (identified by the marker pigment alloxanthin in gut contents) during colder periods, while they preferred small non-filamentous diatoms and green algae (identified mainly by diatoxanthin and lutein, respectively) from May to September. All studied cladoceran species showed highest selectivity towards colonial cyanobacteria (identified by canthaxanthin). For small C. sphaericus, commonly occuring in the pelagic zone of eutrophic lakes, colonial cyanobacteria can be their major food source, supporting their coexistence with cyanobacterial blooms. Pigments characteristic of filamentous cyanobacteria and diatoms (zeaxanthin and fucoxanthin, respectively), algae dominating in Võrtsjärv, were also found in the grazers’ diet but were generally avoided by the crustaceans commonly dominating the zooplankton assemblage. Together these results suggest that the co-occurring small-bodied cyclopoid and cladoceran species have markedly different algal diets and that the cladocera represent the main trophic link transferring cyanobacterial carbon to the food web in a highly eutrophic lake.     

Diversity of phototrophic bacteria in microbial mats from Arctic hot springs (Greenland)

We investigated the genotypic diversity of oxygenic and anoxygenic phototrophic microorganisms in microbial mat samples collected from three hot spring localities on the east coast of Greenland. These hot springs harbour unique Arctic microbial ecosystems that have never been studied in detail before. Specific oligonucleotide primers for cyanobacteria, purple sulfur bacteria, green sulfur bacteria and Choroflexus/Roseiflexus-like green non-sulfur bacteria were used for the selective amplification of 16S rRNA gene fragments. Amplification products were separated by denaturing gradient gel electrophoresis (DGGE) and sequenced. In addition, several cyanobacteria were isolated from the mat samples, and classified morphologically and by 16S rRNA-based methods. The cyanobacterial 16S rRNA sequences obtained from DGGE represented a diverse, polyphyletic collection of cyanobacteria. The microbial mat communities were dominated by heterocystous and non-heterocystous filamentous cyanobacteria. Our results indicate that the cyanobacterial community composition in the samples were different for each sampling site. Different layers of the same heterogeneous mat often contained distinct and different communities of cyanobacteria. We observed a relationship between the cyanobacterial community composition and the in situ temperatures of different mat parts. The Greenland mats exhibited a low diversity of anoxygenic phototrophs as compared with other hot spring mats which is possibly related to the photochemical conditions within the mats resulting from the Arctic light regime.     

Photooxidation of cyanobacteria in natural conditions.

Photodynamic effects were demonstrated and assayed under field conditions in a number of different laboratory strains and pond isolates of cyanobacteria; parameters assayed for resistance to photooxidation were viable count, turbidity of the cyanobacterial suspension, and protein and pigment contents. The effects of density, colonial structure, and internal gas vacuoles on the lethal outcome were investigated. The stability and formation of superoxide dismutase under photooxidative conditions in the field and laboratory were studied in the different strains. An isolate of Microcystis from blooms in ponds exhibited extremely high resistance to photooxidation, which was abolished by exposure to chloramphenicol.     

Can tropical freshwater zooplankton graze efficiently on cyanobacteria?

Zooplankton may at times graze cyanobacteria. However, their top-down effects are considered to be low, particularly in tropical regions dominated by small-size grazers that may be unable to consume efficiently filamentous or colonial species. Recently, cyanobacteria blooms were reported in the Senegal River hydrosystem. We conducted feeding experiments to assess the ability of copepods (Pseudodiaptomus hessei and Mesocyclops ogunnus), cladocerans (Moina micrura and Ceriodaphnia cornuta), and rotifers (Brachionus angularis, B. falcatus, and Keratella sp.) to control different cyanobacteria (Cylindrospermopsis raciborskii, Anabaena solitaria, A. flos-aquae, and Microcystis aeruginosa). None of the zooplankton species ingested M. aeruginosa. Mesocyclops ogunnus did not consume any of the cyanobacteria. Both cladocerans consumed the smallest filaments of cyanobacteria, whereas all the rotifers and P. hessei consumed a broader food-size spectrum. The functional feeding responses suggest that the concentration and size of the filaments are not the sole criteria for food consumption. The high zooplankton community grazing rates, estimated by applying the clearance rates measured in the laboratory to the in situ zooplankton abundance, indicate that grazing by zooplankton potentially constitutes an important controlling factor for the filamentous cyanobacteria in the tropics.     

Flow cytometry sorting of freshwater phytoplankton

We used the flow sorting capacities of a benchtop FACSCalibur flow cytometer to analyze the phytoplankton community of four different aquatic ecosystems. We show that despite the high optical, mechanistic, and hydrodynamic stress for the cells while sorted, most of the targeted populations could be isolated and grew in mixed culture media subsequent to sorting. Forty-five phytoplankton taxa were isolated, including green algae (29 species), cyanobacteria (eight), diatoms (seven), and cryptomonads (one). The isolation success average was high since 80% of the total sorted populations grew successfully and 47% constituted monocultures. It is noteworthy, however, that some groups could not be isolated, as for example colonial cyanobacteria, chrysophytes, euglenophytes, desmids, or dinoflagellates, and some species such as Cryptomonas sp. were very sensitive to the sorting process. It is proposed that flow cytometric analysis of freshwater phytoplankton might be a relevant tool for water managers and could be applied in some specific cases, such as early monitoring of blooming taxa or basic bio-monitorings of key species. The higher isolation average obtained from the flow sorting can also be powerful for the physiological or molecular study of some taxa after their cultivation.     

Testing methods to estimate abundance of Magellanic Penguins Spheniscus magellanicus

Capsule Using simulations in a geographical information system our results showed that systematic sampling with quadrats was the most accurate, precise and cost-effective method to survey Magellanic Penguin colonies.

Aims To determine which sampling method gives better estimations of penguin abundance.

Methods A virtual colony was generated deriving spatial parameters from a real survey and applying Kriging interpolation. Three sampling methods were then applied on this virtual colony: random sampling with quadrats; systematic sampling with quadrats; systematic sampling with fixed-width transects. The estimated abundance for each trial was compared to the abundance of the virtual colony to have a measure of accuracy and precision.

Results Systematic sampling with quadrats estimated penguin abundance better than random or systematic sampling with transects since it achieved 100% accuracy and great precision after sampling only 2.1% of the virtual colony.

Conclusion The use of a simulated colony allowed the comparison of several sampling methods traditionally used in Magellanic Penguin surveys. The results of this study are important in order to standardize sampling protocols for Magellanic Penguins and to have more comparable estimations to detect trends over time. Also, the methodological approach used here could be used to assess sampling methods for other colonial bird species.     

Improved Method for Determining Bacterial Filtration Rates in Zooplankton

Filtration rates were determined for a natural population of zooplankton grazers (Bosmina longirostris [Müll.], Cyclops vicinus vicinus [Ulianine], Acanthodiaptomus denticornis [Wierz.], and Daphnia longispina [Müll.]) by using ³H-labeled bacteria as food for these organisms. There was a relationship between filtration rates of the major zooplankton grazers and the prevailing algal and bacterial composition in the lake water. Low filtration rates were obtained in the presence of colonial and filamentous cyanobacteria. The rapid process of bacterial adhesion to the external organs of grazers can result in an overestimation of filtration rates. By using the simple method presented here, filtration rates, with simultaneous correction for bacterial adhesion, can be quickly determined.