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.

Respiration rates and abundances of dominant copepods of the northern Benguela Current System

Respiration rates of 16 calanoid copepod species from the northern Benguela upwelling system were measured on board RRS Discovery in September/October 2010 to determine their energy requirements and assess their significance in the carbon cycle. Individual respiration rates were standardised to a mean copepod body mass and a temperature regime typical of the northern Benguela Current. These adjusted respiration rates revealed two different activity levels (active and resting) in copepodids C5 of Calanoides carinatus and females of Rhincalanus nasutus, which reduced their metabolism during dormancy by 82% and 62%, respectively. An allometric function (Imax) and an energy budget approach were performed to calculate ingestion rates. Imax generally overestimated the ingestion rates derived from the energy budget approach by >75%. We suggest that the energy budget approach is the more reliable approximation with a total calanoid copepod (mainly females) consumption of 78 mg C m-2 d-1 in neritic regions and 21 mg C m-2 d-1 in oceanic regions. The two primarily herbivorous copepods C. carinatus (neritic) and Nannocalanus minor (oceanic) contributed 83% and 5%, respectively, to total consumption by calanoid copepods. Locally, C. carinatus can remove up to 90% of the diatom biomass daily. In contrast, the maximum daily removal of dinoflagellate biomass by N. minor was 9%. These estimates imply that C. carinatus is an important primary consumers in the neritic province of the northern Benguela system, while N. minor has little grazing impact on phytoplankton populations further offshore. Data on energy requirements and total consumption rates of dominant calanoid copepods of this study are essential for the development of realistic carbon budgets and food-web models for the northern Benguela upwelling system.

Lupane triterpenes from the leaves of the tropical rain forest tree Hopea odorata Roxb. and their cytotoxic activities, supplementary information

Chromatographic fractionation of the cytotoxic n-hexane extract of Hopea odorata Roxb. leaves led to the isolation of eight lupane triterpenes, which constitutes the first report of lupane-type triterpenes from this plant source. Furthermore, 3,30-dioxolup-20(29)-en-28-oic acid (6) was isolated for the first time from a natural source. Their structures were determined on the basis of spectroscopic methods, including 2D NMR analysis, and by comparison of their spectral data with literature values. Complete NMR assignments of the 1H and 13C NMR data were achieved for all compounds. Finally, the cytotoxic activities of the isolated compounds against four human cell lines (PC3, MDA-MB-231, HT-29 and HCT116) was also reported.

pH and calcium change in the microenvironment of a benthic foraminifer (Ammonia sp.) and its size during experiments

Calcareous foraminifera are well known for their CaCO3 shells. Yet, CaCO3 precipitation acidifies the calcifying fluid. Calcification without pH regulation would therefore rapidly create a negative feedback for CaCO3 precipitation. In unicellular organisms, like foraminifera, an effective mechanism to counteract this acidification could be the externalization of H+ from the site of calcification. In this study we show that a benthic symbiont-free foraminifer Ammonia sp. actively decreases pH within its extracellular microenvironment only while precipitating calcite. During chamber formation events the strongest pH decreases occurred in the vicinity of a newly forming chamber (range of gradient about 100 µm) with a recorded minimum of 6.31 (< 10 µm from the shell) and a maximum duration of 7 h. The acidification was actively regulated by the foraminifera and correlated with shell diameters, indicating that the amount of protons removed during calcification is directly related to the volume of calcite precipitated. The here presented findings imply that H+ expulsion as a result of calcification may be a wider strategy for maintaining pH homeostasis in unicellular calcifying organisms.

Ichthyoplankton density and zooplankton biomass in the northern Benguela upwelling system during D356 September 2010

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in September 2010 were determined for 10 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the RRS Discovery D356 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 10 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta(depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during AFR258 in December 2009

IIchthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in December 2009 were determined for 22 stations in the Benguela upwelling system, based on oblique Multinet hauls during the FRS Africana cruise AFR258. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 22 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. Densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).