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).

Absolute and relative abundances of zooplankton taxa in Kongsfjorden during 2002

Seasonal changes in the zooplankton composition of the glacially influenced Kongsfjorden, Svalbard (79°N, 12°E), and its adjacent shelf were studied in 2002. Samples were collected in the spring, summer and autumn in stratified hauls (according to hydrographic characteristics), by means of a 0.180-mm Multi Plankton Sampler. A strong front between the open sea and the fjord waters was observed during the spring, preventing water mass exchange, but was not observed later in the season. The considerable seasonal changes in zooplankton abundance were related to the seasonal variation in hydrographical regime. The total zooplankton abundance during the spring (40-2010 individuals/m**3) was much lower than in the summer and autumn (410-10,560 individuals/m**3). The main factors shaping the zooplankton community in the fjord include: the presence of a local front, advection, the flow pattern and the decreasing depth of the basin in the inner fjord. Presumably these factors regulate the gross pattern of zooplankton density and distribution, and override the importance of biological processes. This study increased our understanding of seasonal processes in fjords, particularly with regard to the strong seasonal variability in the Arctic.

Vertical distribution of relative numbers of the main groups of sound-scattering fishes in the Central Indian Ocean

Findings made in 31 catches with an Isaacs-Kidd midwater trawl in the light (09.00-16.00) and dark (21.00-04.00) periods of a day within a survey area of about 100 sq. miles with approximate center coordinates of 13°S and 78°E have been used to investigate vertical distribution of the main groups of sound-scattering fishes (35 species of the family Myctophidae and 16 species of other families). It has been shown that during daylight hours all fishes sink to depths deeper than 400 m. Data are presented concerning the fish population of night-time sound-scattering layers at depths of 70-150 m and about 400 m and of the daytime ones at depths of about 450 m.

(Table 4) Mesozoolpankton abundance and biomass and relative percentage of key zooplankton groups along the Bulgarian coast, Black Sea

Results of studies during Project of an international expedition onboard R/V Vladimir Parshin in September-October 2005 are presented. Intensive development of Bacillariophyceae and Dynophyceae was recorded in coastal waters of Bulgaria, Turkey, and in the Danube River delta during period of investigations. Increase in algae population was accompanied by rising of chlorophyll a concentration up to 2.0-5.5 µg/l. In the deep water region it did not exceed 0.5 µg/l. Phytoplankton growth rate in the surface water layer varied from 0.1 to 1.0 1/day. This parameter and NO2+NO3 concentration, as well as the silicon concentration were correlative, as was described by the Michaelis-Menten equation. Phytoplankton growth was affected by basic nutrients. Zooplankton grazing varied from 0.10 to 0.69 1/day and average values in different regions varied by 1.5 times. Microalgae size range is one of major factors of grazing regulation. Rate of phytoplankton consumption was decreasing with increasing the largest diatom Pseudosolenia calcar-avis impact on total biomass of nano- and microphytoplankton.

Relative contents of principal diatom groups in suspended matter and surface layer bottom sediments from the Indian Ocean

Data on relative contents of principal diatom groups in suspended matter collected by a separator on the way of the ship and in bottom sediments from different areas of the Indian Ocean are presented in the paper.