Chemical characterization and quantification of the brown algal storage compound laminarin

In search of a meaningful stress indicator for Fucus vesiculosus we found that the often used quantitative determination procedures for the polysaccharide laminarin (beta-1,3-glucan) result in different kind of problems, uncertainties and limitations. This chemical long-term storage form of carbon enables perennial brown algae in seasonally fluctuating ecosystems to uncouple growth from photosynthesis. Because of this high ecological relevance a reliable and precise method for determination and quantification of laminarin is needed. Therefore, a simple, cold water extraction method coupled to a new quantitative liquid chromatography-mass spectrometrical method (LC-MS) was developed. Laminarin was determined in nine out of twelve brown algal species, and its expected typical molar mass distribution of 2000-7000 Da was confirmed. Furthermore, laminarin consisted of a complex mixture of different chemical forms, since fifteen chemical laminarin species with distinct molecular weights were measured in nine species of brown algae. Laminarin concentrations in the algal tissues ranged from 0.03 to 0.86% dry weight (DW). The direct chemical characterization and quantification of laminarin by LC-MS represents a powerful method to verify the biochemical and ecological importance of laminarin for brown algae. Single individuals of Laminaria hyperborea, L. digitata, Saccharina latissima, F. serratus, F. vesiculosus, F. spiralis, Himanthalia elongata, Cystoseira tamariscifolia, Pelvetia canaliculata, Ascophyllum nodosum, Halidrys siliquosa and Dictyota dichotoma were collected in fall (18.11.2013) during spring low tide from the shore of Finavarra, Co. Clare, west coast of Ireland (53° 09' 25'' N, 09° 06' 58'' W). After sampling, the different algae were immediately transported to the lab, lyophilized and sent to the University of Rostock. Laminarin was extracted with cold ultrapure water from the algal samples. Before extraction they were ground to < 1 mm grain size with an analytical mill (Ika MF 10 Basic). The algal material (approx. 1.5 g DW) was extracted in ultrapure water (8 mL) on a shaker (250 rpm) for 5 h. After the addition of surplus ultrapure water (4 mL) and shaking manually, 1 mL of the sample was filter centrifuged (45 µm) at 14,000 rpm (Hettich Mikro 22 R). The slightly viscous supernatant was free of suspended material and converted into a microvial (300 µL) for further analysis. The extracts were analyzed using liquid chromatography-mass spectrometry (LC-MS) analysis (LTQ Velos Pro ion trap spectrometer with Accela HPLC, Thermo Scientific). Laminarin species were separated on a KinetexTM column (2.6 µm C18, 150 x 3 mm). The mobile phase was 90 % ultrapure water and 10 % acetonitrile, run isocratically at a flow rate of 0.2 mL min-1. MS was working in ESI negative ion mode in a mass range of 100 - 4000 amu. Glucose contents were determined after extraction using high-performance liquid chromatography (HPLC). Extracted samples were analyzed in an HPLC (SmartLine, Knauer GmbH) equipped with a SUPELCOGELTM Ca column (30 x 7,8 mm without preColumn) and RI-detector (S2300 PDA S2800). Water was used as eluent at a flow rate of 0.8 mL min-1 at 75 °C. Glucose was quantified by comparison of the retention time and peak area with standard solutions using ChromGate software. Mannitol was extracted from three subsamples of 10-20 mg powdered alga material (L. hyperborea, L. digitata, S. latissima, F. serratus, F. vesiculosus, F. spiralis, H. elongata, P. canaliculata, A. nodosum, H. siliquosa) and quantified, following the HPLC method described by Karsten et al. (1991). For analyzing carbon and nitrogen contents, dried algal material was ground to powder and three subsamples of 2 mg from each alga thalli were loaded and packed into tin cartridges (6×6×12 mm). The packages were combusted at 950 °C and the absolute contents of C and N were automatically quantified in an elemental analyzer (Elementar Vario EL III, Germany) using acetanilide as standard according to Verardo et al. (1990).

Abundance and biomass of dinoflagellates and ciliates at time series station Helgoland Roads, North Sea, 2007-2009

A monitoring programme for microzooplankton was started at the long-term sampling station ''Kabeltonne'' at Helgoland Roads (54°11.30' N; 7°54.00' E) in January 2007 in order to provide more detailed knowledge on microzooplankton occurrence, composition and seasonality patterns at this site and to complement the existing plankton data series. Ciliate and dinoflagellate cell concentration and carbon biomass were recorded on a weekly basis. Heterotrophic dinoflagellates were considerably more important in terms of biomass than ciliates, especially during the summer months. However, in early spring, ciliates were the major group of microzooplankton grazers as they responded more quickly to phytoplankton food availability. Mixotrophic dinoflagellates played a secondary role in terms of biomass when compared to heterotrophic species; nevertheless, they made up an intense late summer bloom in 2007. The photosynthetic ciliate Myrionecta rubra bloomed at the end of the sampling period. Due to its high biomass when compared to crustacean plankton especially during the spring bloom, microzooplankton should be regarded as the more important phytoplankton grazer group at Helgoland Roads. Based on these results, analyses of biotic and abiotic factors driving microzooplankton composition and abundance are necessary for a full understanding of this important component of the plankton.

Late Cretaceous to Eocene calcareous nannofossil distribution near Gubbio, Italy

Using a modified sample preparation technique, we have been able to establish a detailed lower Campanian to upper Eocene nannofossil stratigraphy in the Bottaccione and Contessa Highway sections near Gubbio. Appearance and extinction levels of virtually all the commonly used calcareous nannofossil zonal markers have been recognized and can now be closely correlated with the planktonic foraminifera zonation and the magnetic reversal stratigraphy previously established in these sections. Comparisons with the nannofossil calibrations of the oceanic magnetic anomaly sequence in Deep Sea Drilling Project (DSDP) sites suggest that magmetic Subchrons C17N and C25N are missing in the Bottaccione section. The observed variability of the relative stratigraphic position of most plankton events is confirmed to less than one magnetic subchron. Absolute abundance, paleobiogeographic restriction, and differential preservation render some of the traditionally used biostratigraphic events less reliable than others.

Abundance of mesozooplankton in the SESAME Italian cruise S-IT2 in the Ionian Sea in March 2008

The "SESAME_IT2_ZooAbundance_0-50-100m_SZN" dataset contains data of mesozooplankton species composition and abundance (ind. m-3) from samples collected in the Ionian Sea in the late winter (2-8 March) of 2008 during the SESAME-WP2 cruise IT2. Samples were collected by vertical tows with a closing WP2 net (56 cm diameter, 200 ?m mesh size) in the following depth layers: 100-200 m, 50-100 m, 0-50 m. Sampling was always performed in light hours. A flowmeter was applied to the mouth of the net, however, due to its malfunctioning, the volume of filtered seawater was calculated by multiplying the the area by the height of the sampled layer from winch readings. After collection, each sample was split in two halves (1/2) after careful mixing with graduated beakers. Half sample was immediately fixed and preserved in a formaldehyde-seawater solution (4% final concentration) for species composition and abundance. The other half sample was kept fresh for biomass measurements (data already submitted to SESAME database in different files).Here, only the zooplankton abundance of samples in the upper layers 0-50 m and 50-100 m are presented. The abundance data of the samples in the layer 50-100 m will be submitted later in a separate file. The volume of filtered seawater was estimated by multiplying the the area by the height of the sampled layer from winch readings. Identification and counts of specimens were performed on aliquots (1/20-1/5) of the fixed sample or on the total sample (half of the original sample) by using a graduate large-bore pipette. Copepods were identified to the species level and separated into females, males and juveniles (copepodites). All other taxa were identified at the species level when possible, or at higher taxonomic levels. Taxonomic identification was done according to the most relevant and updated taxonomic literature. Total mesozooplankton abundance was computed as sum of all specific abundances determined as explained above.

Abundance of calcareous nannofossils in ODP Hole 122-761C (Table 1)

A biostratigraphically complete (all nannofossil biozones present) Cretaceous/Tertiary boundary was recovered at Site 761 on the Wombat Plateau, northwest Australian margin, during Ocean Drilling Program Leg 122. A quantitative study of calcareous nannofossils on closely spaced samples across the boundary reveals a rapid change in assemblages in a similar fashion to other Cretaceous/Tertiary boundary sites. Nannofossil species were placed into three categories: Tertiary, Cretaceous, and 'survivors'. The rapid sequential turnover in these assemblages is as follows: Cretaceous species are abruptly replaced at the boundary by opportunistic survivor species, which in turn are abruptly replaced by newly evolved Tertiary taxa. The uppermost Maestrichtian assemblage is distinctly mid-latitudinal with few Micula murus and rare to few Nephrolithus frequens. The nannofossil assemblage immediately above the boundary is dominated by an abundance bloom of Cyclagelosphaera spp. No Thoracosphaera or Braarudosphaera abundance blooms are present as at many other localities. The change from a survivor- to Tertiary-dominated assemblage is coincident with the CPla/CPlb nannofossil subzonal boundary, which is marked by the simultaneous first occurrence of several species including Cruciplacolithus tenuis and C. primus. The latter is found to first occur below C. tenuis in the most complete Cretaceous/Tertiary sections. A hiatus between Subzones CPla and CPlb is interpreted to explain this discrepancy.