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).

Seawater carbonate chemistry, thickness and carbonate elemental composition of the test of juvenile sea urchins in a laboratory experiment

Continuous anthropogenic CO2 emissions to the atmosphere and uptake by the oceans will cause a reduction of seawater pH and saturation state (Omega) of CaCO3 minerals from which marine calcifiers build their shells and skeletons. Sea urchins use the most soluble form of calcium carbonate, high-magnesium calcite, to build their skeleton, spines and grazing apparatus. In order to highlight the effects of increased pCO2 on the test thickness and carbonate elemental composition of juvenile sea urchins and potential differences in their responses linked to the diet, we performed a laboratory experiment on juvenile Paracentrotus lividus, grazing on calcifying (Corallina elongata) and non-calcifying (Cystoseira amentacea, Dictyota dichotoma) macroalgae, under different pH (corresponding to pCO2 values of 390, 550, 750 and 1000 µatm). Results highlighted the importance of the diet in determining sea urchin size irrespectively of the pCO2 level, and the relevance of macroalgal diet in modulating urchin Mg/Ca ratio. The present study provides relevant clues both in terms of the mechanism of mineral incorporation and in terms of bottom-up processes (algal diet) affecting top-down ones (fish predation) in rocky subtidal communities

The Cenomanian/Turronian Boundary Event at ODP Hole 103-641A

Drilling at ODP Site 641 (on the western margin of Galicia Bank, off northwestern Spain) revealed a thin, but pronounced, interval of black shale and gray-green claystone. Our high-resolution study combines the sedimentology, micropaleontology (palynomorphs and others), organic and inorganic geochemistry, and isotopic values of this layer to demonstrate the distinct nature of the sediment and prove that the sequence represents the local sedimentary expression of the global Cenomanian/Turonian Oceanic Anoxic Event (OAE) of Schlanger and Jenkyns (1976), Arthur and Schlanger (1979), and Jenkyns (1980), also called the Cenomanian/Turonian Boundary Event (CTBE). The most striking evidence is that the strong positive d13C excursion characterizing the CTBE sequences in shallow areas can be traced into a pronounced deep-sea expression, thus providing a good stratigraphic marker for the CTBE in various paleosettings. The isotopic excursion at Site 641 coincides with an extremely enriched trace metal content, with values that were previously unknown for the Cretaceous Atlantic. Similar to other CTBE occurrences, the organic carbon content is high (up to 11%) and the organic matter is of dominantly marine origin (kerogen type II). The bulk mineralogy of the CTBE sediments does not differ significantly from the general trend of Cretaceous North Atlantic sediments (dominance of smectite and zeolite with minor amounts of illite and scattered palygorskite, kaolinite, and chlorite); thus, no evidence for either increased volcanic activity nor a drastic climatic change in the borderlands was found. Results from Site 641 are compared with the CTBE section found at Site 398, DSDP Leg 47B (Vigo Seamount at the southern end of the Galicia Bank).

Benthic foraminifera in Lower Cretaceous sediments from various DSDP samples

From the DSDP Legs 1, 11, 13, 17, 25, 27, 32, 36, 41, 43, 44, 50, and 62 the Lower Cretaceous foraminifers have been investigated for biostratigraphical, taxonomical, and palaeoecological purposes. An overview of the cored Lower Cretaceous sections of Leg 1-80 is given. In the Northern Atlantic Ocean characteristic foraminiferal faunas are missing from the Upper Tithonian to the Valanginian due to a marked regression which caused hiatuses. In areas without black shale conditions Valanginian to Barremian medium rich to poor microfaunas with Praedorothia ouachensis (Sigal) of the Praedorothia ouachensis Zone (Valanginian-Hauterivian). The Hauterivian-Aptian interval is characterized by zones of Gavelinella barrerniana, Gaudryina dividens, and Conorotalites aptiensis. During the Albian a world-wide fauna consisting of agglutinated and calcareous foraminifers of the Pseudoclavulina gaultina Zone is established in areas lacking the wide-spread black-shale conditions. The Upper Albian and the Cenomanian are represented by the Gavelinella eenomanica Zone. Some ornamented species of the nodosariids (Citharina, Lenticulina), Gavelinella, Conorotatites, Pleurostomella, Vatvulineria, and Osangularia are of some importance for the biostratigraphy of the Berriasian-Albian interval. The Berriasian to Albian zones introduced for the Tethys and the DSDP by Moullade (1984) could only be of some local importance due to the long stratigraphical range of the foraminiferal species used. In the Indian Ocean an exact stratigraphical age cannot be assigned to the few Neocomian foraminiferal faunas of a cooler sea water (Site 261). These faunas mainly contain primitive agglutinated foraminifers, because in most cases the calcareous tests are dissolved or redeposited. In the Pacific Ocean most of the Berriasian to Aptian microfaunas are of minor biostratigraphical and palaeoecological importance for reasons of poor core recoveries, contaminations or original foraminiferal poverty (black shales). Since the Albian there are somewhat higher-diverse faunas of calcareous and agglutinated foraminifers with index species of the Pseudoclavulina gaultina Zone. As a rule, the boundary Albian/Cenomanian is set by means of planktonic foraminifers because no other foraminifer has its first appearance datum during this interval, except Gavelinella cenornanica. During the Albian very uniform, world-wide foraminiferal faunas without a marked provincialism are obvious.

The hydropolyp fauna of the Red Sea area

46 hydropolyp species of 28 genera and 10 families were sampled during the "Meteor" passage 1964/65 (IIOE) through the Red Sea and its northern and southern exits and on the occasion of several ecological investigations of 29 selected coral reef sections of the central Red Sea and the Gulf of Aqaba. These collections comprise 128 single records of hydropolyp species. Three species and two genera each with one species are doubtful. 25 species, seven genera, one family and one subfamily, together from 49 records have not previously been found in the Red Sea and its exits. Including these newly reported species, the total list increases from 64 species and 112 records to 89 species and 240 single records and 51 additional ones. Scanning microscopical photos, made for the first time for the illustration of the hydropolyps, have been shown to be suitable for a better characterization and diagnosis of the species. Qualified results on the reasons for the horizontal distribution of the species known from the Red Sea area cannot be given because of the low number of samples sporadically distributed through the whole area. In contrast with this fact, the vertical spread of the species sampled seems primarily to be regulated by water exchange and light intensity. For example, four species of hydropolyps are excellent indicators of certain abiotic factors or combinations of them: Gymnangium eximium reacts extremely stenophote-photophobe-rheophil, Eudendrium ramosum moderately stenophote-photophobe-rheophobe, Lytocarpus philippinus moderately stenophote-photophil-rheophil, and Halocordyle disticha var. australis extremely stenophote-photophil but moderately rheophil. Other species have been found throughout all the light zones. Combined with the small size of their colonies their euryphotic behaviour does not allow their use as indicator species.

Dinoflagellate cysts of the Early Cretaceous North Atlantic Ocean

Early Cretaceous dinoflagellate cysts were reinvestigated from nine deep-sea sites of the North and Central Atlantic. In general the zonation scheme developed for the western Central Atlantic (Habib, 1977; Habib and Drugg, 1983 ) can also be applied to the eastern Central Atlantic. Comparison with the probabilistic zonation of Gradstein et al. (1992) show, however, that the first occurrences of the important marker species Druggidium apicopaucicure, Druggidium deflandrei, Druggidium rhabdoreticulatum and Odontochitina operculata appear to occur slightly later in the eastern Central Atlantic in respect to nannofossils and benthic foraminifers. Muderongia neocomica has a shorter stratigraphic range in the eastern Central Atlantic than in the western Central Atlantic.

CO2 and inorganic nutrient enrichment affect the performance of a calcifying green alga and its noncalcifying epiphyte

Ocean acidification studies in the past decade have greatly improved our knowledge of how calcifying organisms respond to increased surface ocean CO2 levels. It has become evident that, for many organisms, nutrient availability is an important factor that influences their physiological responses and competitive interactions with other species. Therefore, we tested how simulated ocean acidification and eutrophication (nitrate and phosphate enrichment) interact to affect the physiology and ecology of a calcifying chlorophyte macroalga (Halimeda opuntia (L.) J.V. Lamouroux) and its common noncalcifying epiphyte (Dictyota sp.) in a 4-week fully crossed multifactorial experiment. Inorganic nutrient enrichment (+NP) had a strong influence on all responses measured with the exception of net calcification. Elevated CO2 alone significantly decreased electron transport rates of the photosynthetic apparatus and resulted in phosphorus limitation in both species, but had no effect on oxygen production or respiration. The combination of CO2 and +NP significantly increased electron transport rates in both species. While +NP alone stimulated H. opuntia growth rates, Dictyota growth was significantly stimulated by nutrient enrichment only at elevated CO2, which led to the highest biomass ratios of Dictyota to Halimeda. Our results suggest that inorganic nutrient enrichment alone stimulates several aspects of H. opuntia physiology, but nutrient enrichment at a CO2 concentration predicted for the end of the century benefits Dictyota sp. and hinders its calcifying basibiont H. opuntia.