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Heavy metal toxicity and bioavailability of dissolved nutrients to a bacterivorous flagellate are linked to suspended particle physical properties.

Many dissolved substances attach easily to sediment particles. In the presence of suspended sediments bioavailability of dissolved substances is therefore, usually reduced and clays are even applied to "wash" natural waters upon pollution. In organisms which feed on food organisms in the size range of these suspended sediment particles, however, bioavailability of such substances may even increase. For microorganisms the interaction with dissolved substances and suspended sediment particles so far has hardly been investigated. We specifically tested: (1) the importance of suspended particles as an uptake route for dissolved substances; and (2) the significance of particle surface properties, i.e. surface load and mineralogy. As a model system we used an axenically cultured strain of a widespread and often abundant flagellate ("Spumella-like" flagellate strain JBM10). We tested the toxicity of cadmium (II) and mercury (II) as well as availability of dissolved organic matter (DOM) in the absence as well as in the presence of different natural clays, i.e. a kaolinite, a montmorillonite, and a mixed clay, and of artificial silicate particles of different surface charge. When applied separately the presence of the heavy metals cadmium and mercury as well as of suspended particles negatively affected the investigated flagellate but nutritive organics supported growth of the investigated flagellate. Toxic stress response comprises behavioral changes including enhanced swimming activity and stress egestion of ingested particles and was generally similar for a variety of different flagellate species. In combination with suspended particles, the respective effect of trace metals and nutritive substances decreased. Regarding the particle quality, cadmium toxicity increased with increasingly negative surface charge, i.e. increasing surface density of silanol groups (Pearson's product moment, P = 0.005). For mercury particle mineralogy still had a significant effect (P < 0.001) but surface load seems to play a minor role and for nutritive organics no significant effect of the investigated particle properties was found. We conclude that: (i) flagellates are as sensitive as higher animals to heavy metal pollution; (ii) suspended particles decrease bioavailability of dissolved substances and ingestion of suspended particles probably play a minor role as uptake route for dissolved substances; and (iii) suspended sediment particle properties, i.e. surface charge and mineralogy, are key factors for the interaction between microorganisms and dissolved substances in the presence of suspended sediments.

Aluminum Silicates↗

A new dihydroxysterol from the marine phytoplankton Diacronema sp.

Diacronema sp. was cultured and its sterols were separated by column chromatography on silica gel. The new sterol 24-ethyl-4alpha-methyl-cholestane-3,20-diol (1) was characterised by NMR and MS spectrometry, as well as (22E)-24-ethyl-4alpha-methyl-5alpha-cholest-22-en-3beta-ol (2) and beta-sitosterol, the major components of the sterol fractions. Neither the biosynthetic origin of the new dihydroxysterol nor its role in the biochemistry of Diacronema is known.

Animals↗

Multiplex PCR allows simultaneous detection of pathogens in ships' ballast water.

There is enormous potential for global transfer of microorganisms, including pathogens, in ships' ballast water. We contend that a major advancement in the study of ballast-water microorganisms in particular, and of aquatic pathogens in general, will be expedited sample analysis, such as provided by the elegant technology of DNA microarrays. In order to use DNA microarrays, however, one must establish the appropriate conditions to bind target sequences in samples to multiple probes on the microarrays. We conducted proof-of-concept experiments to optimize simultaneous detection of multiple microorganisms using polymerase chain reaction (PCR) and Southern hybridization. We chose three target organisms, all potentially found in ballast water: a calicivirus, the bacterium Vibrio cholerae, and the photosynthetic protist Aureococcus anophagefferens. Here, we show simultaneous detection of multiple pathogens is possible, a result supporting the promising future use of microarrays for simultaneous detection of pathogens in ballast water.

Blotting, Southern↗

Energy--dependent bacterivory in Ochromonas minima--a strategy promoting the use of substitutable resources and survival at insufficient light supply.

Phagotrophy and competitive ability of the mixotrophic Ochromonas minima were investigated in a three-factorial experiment where light intensity (low: 1.0 micromol m(-2)s(-1) and high: 60 micromol m(-2)s(-1) PPFD), nutrient concentration (ambient: 7.0 micromolNl(-1), 0.11 micromol P l(-1) and enriched: 88 micromol N l(-1), 6.3 micro mol P l(-1)) and DOC supply (without and with enrichment, 250 micromol C l(-1)) were manipulated. Ochromonas minima and bacterial abundance were monitored for 12 days. We found significant and interacting effects of light and nutrients on Ochromonas minima growth rate and abundance. At high light intensity, nutrient enrichment resulted in increased growth rates and population sizes. In contrast, reduced growth rates and population sizes were observed for nutrient enrichment when light intensity was low. Although, Ochromonas minima was able to ingest bacteria under both high and low light conditions, it grew only when light intensity was high. At high light intensity, Ochromonas minima grew exponentially under nutrient conditions that would have been limiting for photoautotrophic microalgae. In non-enriched low light treatments, Ochromonas minima populations survived, probably by using background DOC as an energy source, indicating that this ability can be of relevance for natural systems even when DOC concentrations are relatively low. When competing with photoautotrophic microalgae, the ability to grow under severe nutrient limitation and to survive under light limitation should be advantageous for Ochromonas minima.

Bacteria↗

Diverse eukaryotes have retained mitochondrial homologues of the bacterial division protein FtsZ.

Mitochondrial fission requires the division of both the inner and outer mitochondrial membranes. Dynamin-related proteins operate in division of the outer membrane of probably all mitochondria, and also that of chloroplasts--organelles that have a bacterial origin like mitochondria. How the inner mitochondrial membrane divides is less well established. Homologues of the major bacterial division protein, FtsZ, are known to reside inside mitochondria of the chromophyte alga Mallomonas, a red alga, and the slime mould Dictyostelium discoideum, where these proteins are likely to act in division of the organelle. Mitochondrial FtsZ is, however, absent from the genomes of higher eukaryotes (animals, fungi, and plants), even though FtsZs are known to be essential for the division of probably all chloroplasts. To begin to understand why higher eukaryotes have lost mitochondrial FtsZ, we have sampled various diverse protists to determine which groups have retained the gene. Database searches and degenerate PCR uncovered genes for likely mitochondrial FtsZs from the glaucocystophyte Cyanophora paradoxa, the oomycete Phytophthora infestans, two haptophyte algae, and two diatoms--one being Thalassiosira pseudonana, the draft genome of which is now available. From Thalassiosira we also identified two chloroplast FtsZs, one of which appears to be undergoing a C-terminal shortening that may be common to many organellar FtsZs. Our data indicate that many protists still employ the FtsZ-based ancestral mitochondrial division mechanism, and that mitochondrial FtsZ has been lost numerous times in the evolution of eukaryotes.

Amino Acid Sequence↗

High diversity of the 'Spumella-like' flagellates: an investigation based on the SSU rRNA gene sequences of isolates from habitats located in six different geographic regions.

We isolated 28 strains of 'Spumella-like' flagellates from different freshwater and soil habitats in Austria, People's Republic of China, Nepal, New Zealand, Uganda, Kenya, Tanzania and Hawaii by use of a modified filtration-acclimatization method. 'Spumella-like' flagellates were found in all of the samples and were often among the dominant bacterivorous flagellates in the respective environments. The small subunit ribosomal RNA (SSU rRNA) gene sequence of the isolates was determined and aligned with previously published sequences of members belonging to the Chrysophyceae sensu stricto. Phylogenetic analysis of the 28 new sequences confirmed their position within the Chrysophyceae sensu stricto and positioned them within different clades. Most of the sequences grouped within clade C and formed several subclusters separated from each other by green taxa including flagellates belonging to Ochromonas, Dinobryon, Poterioochromonas and others. All soil isolates clustered together (subcluster C1) with the soil strain Spumella elongata and the undescribed soil strain 'Spumella danica'. Aquatic isolates were affiliated with at least two branches (C2 and C3). Sequence similarity to the closest related member of the Chrysophyceae ranged between 92% and 99.6%, sequence divergence among the 'Spumella-like' flagellates was as high as 10%. We conclude that (i) the 'Spumella-like' flagellates are a diverse group both in terms of sequence dissimilarity between isolates and in terms of the number of genotypes, (ii) Spumella and Ochromonas are polyphyletic, and (iii) based on the SSU rRNA gene no biogeographical restriction of certain branches could be observed even though different ecotypes may be represented by the same genotype.

Base Sequence↗

Evidence for geographic isolation and signs of endemism within a protistan morphospecies.

The possible existence of endemism among microorganisms resulting from and preserved by geographic isolation is one of the most controversial topics in microbial ecology. We isolated 31 strains of "Spumella-like" flagellates from remote sampling sites from all continents, including Antarctica. These and another 23 isolates from a former study were characterized morphologically and by small-subunit rRNA gene sequence analysis and tested for the maximum temperature tolerance. Only a minority of the Spumella morpho- and phylotypes from the geographically isolated Antarctic continent follow the worldwide trend of a linear correlation between ambient (air) temperature during strain isolation and heat tolerance of the isolates. A high percentage of the Antarctic isolates, but none of the isolates from locations on all other continents, were obligate psychrophilic, although some of the latter were isolated at low ambient temperatures. The drastic deviation of Antarctic representatives of Spumella from the global trend of temperature adaptation of this morphospecies provides strong evidence for geographic transport restriction of a microorganism; i.e., Antarctic protistan communities are less influenced by transport of protists to and from the Antarctic continent than by local adaptation, a subtle form of endemism.

Acclimatization↗

Succession and regulation factors of small eukaryote community composition in a lacustrine ecosystem (Lake Pavin).

The structure and dynamics of small eukaryotes (cells with a diameter less than 5 microm) were studied over two consecutive years in an oligomesotrophic lake (Lake Pavin in France). Water samples were collected at 5 and 30 m below the surface; when the lake was stratified, these depths corresponded to the epilimnion and hypolimnion. Changes in small-eukaryote structure were analyzed using terminal restriction fragment length polymorphism (T-RFLP) and cloning and sequencing of the 18S rRNA genes. Terminal restriction fragments from clones were used to reveal the dominant taxa in T-RFLP profiles of the environmental samples. Spumella-like cells (Chrysophyceae) did not dominate the small eukaryote community identified by molecular techniques in lacustrine ecosystems. Small eukaryotes appeared to be dominated by heterotrophic cells, particularly Cercozoa, which represented nearly half of the identified phylotypes, followed by the Fungi-LKM11 group (25%), choanoflagellates (10.3%) and Chrysophyceae (8.9%). Bicosoecida, Cryptophyta, and ciliates represented less than 9% of the community studied. No seasonal reproducibility in temporal evolution of the small-eukaryote community was observed from 1 year to the next. The T-RFLP patterns were related to bottom-up (resources) and top-down (grazing) variables using canonical correspondence analysis. The results showed a strong top-down regulation of small eukaryotes by zooplankton, more exactly, by cladocerans at 5 m and copepods at 30 m. Among bottom-up factors, temperature had a significant effect at both depths. The concentrations of nitrogenous nutrients and total phosphorus also had an effect on small-eukaryote dynamics at 5 m, whereas bacterial abundance and dissolved oxygen played a more important structuring role in the deeper zone.

Animals↗

Cyanobacterial blooms and the presence of cyanotoxins in small high altitude tropical headwater reservoirs in Kenya.

The phytoplankton community in three small (0.065-0.249 km2) reservoirs in the stepped plateau landscape in the Kinangop area above the Rift Valley floor in Kenya were studied between 1998 and 2000. Approximately 70 species of phytoplankton were identified. The community was dominated by chlorophytes, cyanobacteria and chrysophytes. Diatoms were rare. The phytoplankton assemblage was frequently dominated by cyanobacteria in the dry season. The phytoplankton assemblage transformed to a mixture of cyanobacteria, chlorophytes and chrysophytes at the onset of the long rains, and mixture of cyanobacteria and chlorophytes after the long rains. Thereafter the phytoplankton assemblage consisted mainly of a mix of cyanobacteria and chrysophytes until the onset of the short rains when cyanobacterial dominance re-emerged. The most common phytoplankton species included Microcystis spp., Botryococcus braunii, Ceratium hirundinella, Anabaena spp. and Euglena viridis. The dry season cyanobacterial blooms produced cyanotoxins that included microcystin and endotoxins. The concentrations were well above the recommended safe limits for drinking water. The patterns of cyanotoxin production showed that the growth of the toxin-producing cyanobacteria was regulated by water temperature, pH and nutrients. The appearance of cyanotoxins in the small reservoirs is a serious public health issue in rural Kenya because such reservoirs are key sources of water for humans, livestock and wildlife.

Altitude↗