Decrease in volume and density of foraminiferal shells with progressing ocean acidification.

Rapid increases in anthropogenic atmospheric CO2 partial pressure have led to a decrease in the pH of seawater. Calcifying organisms generally respond negatively to ocean acidification. Foraminifera are one of the major carbonate producers in the ocean; however, whether calcification reduction by ocean acidification affects either foraminiferal shell volume or density, or both, has yet to be investigated. In this study, we cultured asexually reproducing specimens of Amphisorus kudakajimensis, a dinoflagellate endosymbiont-bearing large benthic foraminifera (LBF), under different pH conditions (pH 7.7-8.3, NBS scale). The results suggest that changes in seawater pH would affect not only the quantity (i.e., shell volume) but also the quality (i.e., shell density) of foraminiferal calcification. We proposed that pH and temperature affect these growth parameters differently because (1) they have differences in the contribution to the calcification process (e.g., Ca2+-ATPase and Ω) and (2) pH mainly affects calcification and temperature mainly affects photosynthesis. Our findings also suggest that, under the IPCC RCP8.5 scenario, both ocean acidification and warming will have a significant impact on reef foraminiferal carbonate production by the end of this century, even in the tropics.

A method for stable carbon isotope measurement of underivatized individual amino acids by multi-dimensional high-performance liquid chromatography and elemental analyzer/isotope ratio mass spectrometry.

RATIONALE: To achieve better precision and accuracy for δ13 C analysis of individual amino acids (AAs), we have developed a new analytical method based on multi-dimensional high-performance liquid chromatography (HPLC) and elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). Unlike conventional methods using gas chromatography, this approach omits pre-column chemical derivatization, thus reducing systematic errors associated with the isotopic measurement. METHODS: The separation and isolation of individual AAs in a standard mixture containing 15 AAs and a biological sample, spear squid (Heterololigo bleekeri) were performed. AAs were isolated using an HPLC system equipped with a reversed-phase column and a mixed-mode column and collected using a fraction collector. After the chromatographic separation and further post-HPLC purification, the δ13 C values of AAs were measured by EA/IRMS. RESULTS: The complete isolation of all 15 AAs in the standard mixture was achieved. The δ13 C values of these AAs before and after the experiment were in good agreement. Also, 15 AAs in the biological sample, H. bleekeri, were successfully measured. The δ13 C values of AAs in H. bleekeri varied by as much as 30‰ with glycine being most enriched in13 C. CONCLUSIONS: The consistency between the δ13 C values of reference and processed AAs demonstrates that the experimental procedure generates accurate δ13 C values unaffected by fractionation effects and contamination. This method is therefore suitable for δ13 C analysis of biological samples with higher precision than conventional approaches. We propose this new method as a tool to measure δ13 C values of AAs in biological, ecological and biogeochemical studies.

Lithium, magnesium and sulfur purification from seawater using an ion chromatograph with a fraction collector system for stable isotope measurements.

We describe the mass descrimination and validation of an offline method for purification of Li, Mg and S with an ion chromatograph coupled to an automated fraction collector for use prior to stable isotope measurements. Significant sub-fraction mass fractionation was observed for both the Li and the Mg stable isotope ratios. The lighter Li and heavier Mg isotopes were preferentially retained by the column, resulting in 7Li/6Li and 26Mg/24Mg biases up to 85.8‰ and 0.95‰, respectively. The isotopic compositions of Li, Mg, and S separated from seawater were δ7LiL-SVEC = +30.9‰, δ26MgDSM3 = -0.83 ± 0.10‰, and δ34SVCDT = +19.4 ± 0.6‰; each chromatographic peak was completely recovered, and the results were in good agreement with the published values regardless of whether or not chemical suppressor was used. The purification method enables multi-isotope analysis of a sample using various mass spectrometry techniques, such as multiple-collector inductively coupled plasma and thermal ionization mass spectrometry.

Identification of the chemical form of sulfur compounds in the Japanese pink coral (Corallium elatius) skeleton using µ-XRF/XAS speciation mapping.

The distributions and chemical forms of sulfur compounds in the skeleton of Japanese pink coral (Corallium elatius) were investigated using X-ray spectroscopic techniques combined with micro-focused soft X-ray radiation. Microscopic X-ray fluorescence/soft X-ray photoabsorption (µ-XRF/XAS) speciation mapping clarified that sulfate is the primary species in the coral skeleton, with minor amounts of organic sulfur, whereas both sulfate and organic sulfur coexist in coenenchyme. Analysis of the post-edge region of the XAS spectra confirmed that sulfate ions in the coral skeleton are mainly in the form of gypsum-like inorganic sulfate substituting for the carbonate ions in the calcite skeleton. The sulfate concentration was negatively correlated with the magnesium concentration and positively correlated with that of phosphorus. Speciation mapping of sulfate in the coral skeleton showed clear fluctuations with sulfate concentrations being higher at dark bands, whereas the small amount of organic sulfur had unclear dark/bright bands. These results suggest that the little organic sulfur that is present is contained in the organic matter embedded in the biocrystal of coral skeleton.

Magnesium K-edge XANES spectroscopy of geological standards.

Magnesium K-edge X-ray absorption near-edge structure (XANES) spectra have been investigated to develop a systematic understanding of a suite of Mg-bearing geological materials such as silicate and carbonate minerals, sediments, rocks and chemical reagents. For the model compounds the Mg XANES was found to vary widely between compounds and to provide a fingerprint for the form of Mg involved in geologic materials. The energy positions and resonance features obtained from these spectra can be used to specify the dominant molecular host site of Mg, thus shedding light on Mg partitioning and isotope fractionation in geologic materials and providing a valuable complement to existing knowledge of Mg geochemistry.

Microscopic observation of symbiotic and aposymbiotic juvenile corals in nutrient-enriched seawater.

Symbiotic and aposymbiotic juvenile corals, which were grown in the laboratory from the gametes of the scleractinian coral Acropora digitifera and had settled down onto plastic culture plates, were observed with a microscope under different nutrient conditions. The symbiotic corals successfully removed the surrounding benthic microalgae (BMA), whereas the aposymbiotic corals were in close physical contact with BMA. The areal growth rate of the symbiotic corals was significantly higher than that of the aposymbiotic corals. The addition of nutrients to the culture seawater increased the chlorophyll a content in the symbiotic coral polyps and enhanced the growth of some of the symbiotic corals, however the average growth rate was not significantly affected, most likely because of the competition with BMA. The comparison between the symbiotic and aposymbiotic juvenile corals showed that the establishment of a symbiotic association could be imperative for post-settlement juvenile corals to survive in high-nutrient seawater.

Effects of acidified seawater on coral calcification and symbiotic algae on the massive coral Porites australiensis.

We investigated the effect of acidified seawater on calcification and symbiotic algae (zooxanthellae density, chlorophyll content per single algal cell, fluorescence yield (Fv/Fm)) on a massive coral, Porites australiensis, a common species in the Ryukyu Archipelago of Japan. We found that acidified seawater significantly decreased the calcification and fluorescence yield, but did not affect zooxanthellae density and chlorophyll content per single algal cell. This indicates low levels of photoacclimation to acidified seawater in this species, and this is contrary to the findings of previous studies of Acropora species. A significant correlation between calcification and fluorescence yield was observed, indicating the presence of a strong relationship between calcification and algal photosynthesis. Our results indicate that endosymbiont photosynthetic dysfunction may enhance the decrease of coral calcification in future acidified ocean conditions. Calcification and fluorescence yield among colonies clearly differed, showing that the response to acidified seawater is highly variable among colonies in natural coral populations.

Mg isotope fractionation in biogenic carbonates of deep-sea coral, benthic foraminifera, and hermatypic coral.

High-precision Mg isotope measurements by multiple collector inductively coupled plasma mass spectrometry were applied for determinations of magnesium isotopic fractionation of biogenic calcium carbonates from seawater with a rapid Mg purification technique. The mean δ(26)Mg values of scleractinian corals, giant clam, benthic foraminifera, and calcite deep-sea corals were -0.87‰, -2.57‰, -2.34‰, and -2.43‰, suggesting preferential precipitation of light Mg isotopes to produce carbonate skeleton in biomineralization. Mg isotope fractionation in deep-sea coral, which has high Mg calcite skeleton, showed a clear temperature (T) dependence from 2.5 °C to 19.5 °C: 1,000 × ln(α) = -2.63 (±0.076) + 0.0138 (±0.0051) × T(R(2) = 0.82, p < 0.01). The δ(26)Mg values of large benthic foraminifera, which are also composed of a high-Mg calcite skeleton, can be plotted on the same regression line as that for deep-sea coral. Since the precipitation rates of deep-sea coral and benthic foraminifera are several orders of magnitude different, the results suggest that kinetic isotope fractionation may not be a major controlling factor for high-Mg calcite. The Mg isotope fractionation factors and the slope of temperature dependence from deep-sea corals and benthic foraminifera are similar to that for an inorganically precipitated calcite speleothem. Taking into account element partitioning and the calcification rate of biogenic CaCO(3), the similarity among inorganic minerals, deep-sea corals, and benthic foraminiferas may indicate a strong mineralogical control on Mg isotope fractionation for high-Mg calcite. On the other hand, δ(26)Mg in hermatypic corals composed of aragonite has been comparable with previous data on biogenic aragonite of coral, sclerosponges, and scaphopad, regardless of species differences of samples.

Permanent El Niño during the Pliocene warm period not supported by coral evidence.

The El Niño/Southern Oscillation (ENSO) system during the Pliocene warm period (PWP; 3-5 million years ago) may have existed in a permanent El Niño state with a sharply reduced zonal sea surface temperature (SST) gradient in the equatorial Pacific Ocean. This suggests that during the PWP, when global mean temperatures and atmospheric carbon dioxide concentrations were similar to those projected for near-term climate change, ENSO variability--and related global climate teleconnections-could have been radically different from that today. Yet, owing to a lack of observational evidence on seasonal and interannual SST variability from crucial low-latitude sites, this fundamental climate characteristic of the PWP remains controversial. Here we show that permanent El Niño conditions did not exist during the PWP. Our spectral analysis of the δ(18)O SST and salinity proxy, extracted from two 35-year, monthly resolved PWP Porites corals in the Philippines, reveals variability that is similar to present ENSO variation. Although our fossil corals cannot be directly compared with modern ENSO records, two lines of evidence suggest that Philippine corals are appropriate ENSO proxies. First, δ(18)O anomalies from a nearby live Porites coral are correlated with modern records of ENSO variability. Second, negative-δ(18)O events in the fossil corals closely resemble the decreases in δ(18)O seen in the live coral during El Niño events. Prior research advocating a permanent El Niño state may have been limited by the coarse resolution of many SST proxies, whereas our coral-based analysis identifies climate variability at the temporal scale required to resolve ENSO structure firmly.

Chemistry of fly ash and cyclone ash leachate from waste materials and effects of ash leachates on bacterial growth, nitrogen-transformation activity, and metal accumulation.

The effects of waste ash leachates on soil microorganism were evaluated along with a chemical characterization of ash leachates. Thirty fly ash samples and cyclone ash samples obtained from the incineration of municipal solid waste, plastic waste, and construction waste were used. Twenty-one and 22 samples inhibited N transformation activity of soil microorganism and growth of Bacillus subtilis, respectively. On the other hand, 11 and 18 samples stimulated bacterial activity and growth, respectively, at low concentrations. Generally, cyclone ash contained a smaller amount of toxic metals than fly ash. Our results suggest that cyclone ash can be further studied for reuse, perhaps as a soil amendment. Pb was found to be highly accumulated in B. subtilis cells, and should be carefully monitored when waste ash is reused in the environment.

Distribution of pesticides and bisphenol A in sediments collected from rivers adjacent to coral reefs.

To investigate the deteriorating health of coral reefs in Okinawa, Japan, natural sediment samples were analyzed for diuron, Irgarol 1,051, chlorpyrifos, and bisphenol A (BPA) which are hazardous to corals. Samples were analyzed by solid-phase extraction (SPE) followed by high-performance liquid chromatography with tandem mass spectrometry (LC-MS-MS). Although diuron and chlorpyrifos usage is only well recorded for farms and not for cities, these chemicals were detected in both rural and urban areas. Additionally, diuron concentration in urban areas was in some cases higher than in rural areas, which might be caused by greater consumption of these chemicals in home gardens in city areas. Irgarol 1051 was detected in downstream river areas, which are situated far from the source sites such as pier or fishery harbor (0.6-3.2 km). This result suggested that Irgarol 1051 could be transported from the river mouths to the sampling sites during flood tides. High BPA concentrations were associated with urban areas (<1.2-22.0 microg kg(-1)), while low concentrations were associated with rural areas (nd-6.8 microg kg(-1)). The river sediments under study are delivered to coral reefs in large quantity through runoff caused by typhoons and other heavy rains. The highly hazardous chemicals are carried into coral reefs on these sediments. Therefore, these hazardous chemical substances may already be influencing the coral reefs.

Arsenic resistance and removal by marine and non-marine bacteria.

Arsenic resistance and removal was evaluated in nine bacterial strains of marine and non-marine origins. Of the strains tested, Marinomonas communis exhibited the second-highest arsenic resistance with median effective concentration (EC(50)) value of 510 mg As l(-1), and was capable of removing arsenic from culture medium amended with arsenate. Arsenic accumulation in cells amounted to 2290 microg As g(-1) (dry weight) when incubated on medium containing 5 mg As l(-1) of arsenate. More than half of the arsenic removed was related to metabolic activity: 45% of the arsenic was incorporated into the cytosol fraction and 10% was found in the lipid-bound fraction of the membrane, with the remaining arsenic considered to be adsorbed onto the cell surface. Potential arsenic resistance and removal were also examined in six marine and non-marine environmental water samples. Of the total bacterial colony counts, 28-100% of bacteria showed arsenic resistance. Some of the bacterial consortia, especially those from seawater enriched with arsenate, exhibited higher accumulated levels of arsenic than M. communis under the same condition. These results showed that arsenic resistant and/or accumulating bacteria are widespread in the aquatic environment, and that arsenic-accumulating bacteria such as M. communis are potential candidates for bioremediation of arsenic contaminated water.

Distribution and temporal changes of lead in the surface seawater in the western Pacific and adjacent seas derived from coral skeletons.

We determined Pb contents together with Pb isotopic compositions in coral skeletons (Porites spp.) collected from the western Pacific and adjacent seas to examine distribution of Pb in sea surfaces. Temporal records of Pb were also investigated using coral skeletons from Hainan and Ogasawara Island, located in China and Japan, respectively. The spatial distribution of Pb showed a clear dilution pattern of Pb from Asian continent to the open ocean. Also the similar trend was found in the Java Sea from Jakarta to the offshore. In addition to the spatial distribution, Pb contents in Ogasawara coral have gradually increased during last 108 years. It may be attributed to Pb emission mainly from industrial activities in Asian countries. Hainan coral, which provided 10 years record of Pb, showed a remarkable decline around 1997 probably due to the partial introduction of unleaded gasoline in China.

Endocrine disrupter nonylphenol and bisphenol A contamination in Okinawa and Ishigaki Islands, Japan--within coral reefs and adjacent river mouths.

Certain chemicals possess the potential to modulate endocrine systems, and thereby interfere with reproduction and developmental processes in the wild. We analyzed endocrine disrupters nonylphenol (NP) and bisphenol A (BPA) levels at various sites in Okinawa and Ishigaki Islands, Japan. River-water samples showed undetectable to low concentrations of NP and BPA at most of the sites investigated. However, an appreciable amount of BPA was detected in sediments at one coral reef site. In addition, significant numbers of river sediment samples showed appreciable amounts of NP and BPA. Most of the sampling sites for this study are located within a distance of 1 km from the coral reefs, which are under influence of river-waters to a variable extent. Therefore, influence of endocrine disrupters may have already begun on adjacent coral reefs. Both endocrine disrupters were positively correlated with human population densities, but not with the contents of red soil generated by farm land reformation. Therefore, it is concluded that NP and BPA pollution is a consequence of human waste discharge, both domestic and industrial, and not by agricultural activities.

Coral skeletal tin and copper concentrations at Pohnpei, Micronesia: possible index for marine pollution by toxic anti-biofouling paints.

We present 40 year-long skeletal chronologies of tin (Sn) and copper (Cu) from an annually-banded coral (Porites sp.) collected from Pohnpei Island, Micronesia (western equatorial Pacific). Both the elements are present in antifouling marine paints and are released inadvertently into ambient seawater. Especially, Sn has often been used in the form of tributyltin (TBT). Based on a stepwise pretreatment examination, Sn and Cu both inside and outside the aragonite lattice of the coral skeleton show a potential for providing marine pollution indicators. High values of extra-skeletal Cu/Ca and Sn/Ca atomic ratios were found between late 1960s and late 1980s during a period of active use of TBT-based antifouling paints worldwide. However, a significant decrease in both the ratios in the beginning of 1990s can be attributed to regulation of the use of TBT on cargo ships by countries such as the USA, Japan and Australia.