Electrospray ionization-quadrupole ion trap mass spectrometry (ESI-QITMS), either in positive- or in negative-ion mode, has been used to establish the chemical structures (chain length, degree of unsaturation, positional distribution) of the fatty acids attached to the primary (sn-1) and secondary (sn-2) hydroxyl groups of the glycerol moiety of natural monogalactosyl- (MGDG) and digalactosyldiacylglycerols (DGDG), isolated from the freshwater dinoflagellate Glenodinium sanguineum and from a marine diatom belonging to the genus Chaetoceros. Fragmentation by collision-induced dissociation of a single component in MGDG and DGDG mixtures, separated by high-performance liquid chromatography (HPLC) and detected on-line by tandem positive-ion ESI-MS, leads to a clear-cut determination of the positional distribution of the sn-glycerol-bound fatty acyl chains. Reversed-phase liquid chromatography allowed a partial resolution of the component mixture before ESI-MS/MS analysis. These results were validated by comparison with ESI-MS data obtained for the sn-2 lysoglyceroglycolipids synthesized via regiospecific enzymatic hydrolysis of the corresponding diacylglycerols by Rhizopus arrhizus lipase.

The depth-distribution of epilithic diatoms in a carbonate meromictic lake (south-eastern Alps), characterized by marked water-level fluctuations, was investigated. Fixed stations were placed along a depth-profile at intervals of 2-3 m and sampled throughout the year using scuba diving. Diatom analysis included quantification of living cells (biovolume calculations) and the estimation of fucoxanthin concentration, which, coupled with the analysis of digested material, permitted taxonomic determination at specific or subspecific level. Multivariate analyses indicated the following variables to be significant: water-level fluctuations, photosynthetically active radiation, silica and nitrates. Diatoms had distinct depth-distributions. Quantification of densities, biovolumes and Chl a, and physiological (senescence index, fucoxanthin), functional (photosynthetic efficiency), and community structure (diversity) parameters allowed the definition of three depth-distribution zones (shallow, mid-depth and deep). The different diatom communities in these zones were confirmed by ANOSIM (analysis of similarity). The shallow, mid-depth, and deep zone were characterized by disturbance due to water-level fluctuations, high stability with favourable growth conditions and severe light limitation, respectively. Community composition and diversity depth-distribution features remained relatively stable throughout the year, while marked changes in benthic diatom biovolumes were modulated by competition for light with phytoplankton. Fucoxanthin was strongly correlated with epilithic diatom biovolumes. The results indicate that depth-distribution patterns should be considered when performing lake diatom biodiversity inventories and integrity evaluations because water-level fluctuations are likely to increase in many lakes due to increased withdrawal of water and climate change in the future.

Ochrobactrum anthropi is a gram-negative bacillus recognized as a human opportunist pathogen isolated in clinical specimens and not of clinical significance. We report a new aspect of this bacterium, that it has been isolated from activated sludge. In fact, it is able to grow on atrazine (2-chloro-4-ethylamino-6-isopropyl-amines-triazine) by utilizing it as the only source of carbon. Our results show that atrazine (0.03 g/liter) causes a dramatical increase in the degree of saturation of membrane fatty acids. Analysis and identification of bacterial fatty acids were performed by gas chromatography and gas chromatography-mass spectrometry techniques.

In Lake Tovel, an oligotrophic and weakly stratified lake, the dinoflagellate Borghiella dodgei Moestrup, Hansen et Daugbjerg, showed a peculiar spatial-temporal pattern with highest abundances in the bottom of the shallow side bay (4 m) along with remarkable abundance variations fromyear to year. We investigated B. dodgei's growth in laboratory cultures and related results to their implication for bloom formation. B. dodgei was cultivated under different temperature, nutrient and light conditions. Growth rates, cell biovolume, cyst formation and pigment and mycosporine-like amino acids (MAAs) concentrations were determined. Experiments showed that this alga (i) had higher growth rates at low temperatures (\7°C) and high irradiance levels (*250 lmol m -2 s -1 ), (ii) produced higher yields with organic supplements such as peptone, (iii) did not grow in the dark even with organic supplements, (iv) survived for long periods without a light source, (v) synthesised MAAs, (vi) showed an increase in cell volume with nutrient shortage and increasing temperatures ([7°C) and (vii) had high encystment rates with temperatures [7°C. These laboratory fingerprints allowed us to construct a theoretical model defining the species' niche. Borghiella needed a mixture of low temperatures, high irradiance levels and sufficient quantities of dissolved organic nitrogen to form blooms. Such a strict combination was probably a transient situation and occurred in oligotrophic Lake Tovel only in early summers followed by heavy spring rains.

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Liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) has been employed to identify carotenoid esters present in raw organic extracts of pigmented freshwater microalgae and to gain structural information on these compounds. In particular, acyl carotenoid derivatives of Haematococcus pluvialis and Euglena sanguinea have been characterised by tandem mass spectrometry (MS/MS) in a quadrupole ion trap. ESI-MS/MS allows recognition of the presence of carotenoid esters in complicated mixtures without any initial chromatographic work-up and without the need to use UV-Vis photo-diode array (PDA) detectors. Product ion scans of the [M + Na]+ ion lead to known neutral losses of the C7H8 and C8H10 residues from the conjugated polyene moiety of the carotenoid unit, that permit the unambiguous identification of the carotenoid itself. These structurally relevant ions are not observed in positive or negative ion APCI (atmospheric pressure chemical ionisation) mass spectra. Moreover, the several product ions observed in positive and/or negative ion ESI-MS/MS not only are a diagnostic signature of the main structural features of the acyl chains such as length, position and unsaturation, but also display the nominal mass of the parent xanthophyll. Our methodology has been validated (i) by using esters of astaxanthin obtained from off-line purification of the H. pluvialis extracts and structurally elucidated through proton nuclear magnetic resonance (1H-NMR) spectroscopy and (ii) by product analysis of esters by alkaline hydrolysis. The characterisation of the unknown carotenoid esters of E. sanguinea is a demonstration of the capabilities of this methodology.

Hemivannusal (6), a bicyclic sesquiterpenoid with an unprecedented skeleton, has been isolated from the marine ciliate Euplotes vannus (strain TB6). The relative configuration of 6 has been established through an extended conformational search performed by molecular mechanics and refined through ab initio quantum chemical calculations. Another strain (CM1) of the same morphospecies has been found to produce the linear prevannusadial A (7) and the monocyclic prevannusadial B (8), which could represent key intermediates for the biosynthesis both of hemivannusal (6) and of the C30 terpenoids vannusals A and B (5a and 5b), the latter pair being metabolites isolated from conspecific strains (Sil21 and

New pigments with an unprecedented skeleton, named keronopsamides A-C, were isolated from the marine ciliate Pseudokeronopsis riccii. The structure of the most abundant secondary metabolite, keronopsamide A, was established through extended Nuclear Magnetic Resonance (NMR) analysis and mass spectrometric (MS) data obtained by using electrospray (ESI) and matrix assisted laser desorption (MALDI) ionizations. Structures of the minor analogues (keronopsamide B and C) were inferred from 1 H NMR, chemical correlation, and LC-MS measurements. The analysis of NOE dipolar couplings and quantum chemical calculations,