Using Museum collections to assess the impact of industrialization on mussel (Mytilus edulis) calcification.

Mytilus edulis is a commercially and ecologically important species found along the east coast of the United States. Ecologically, M. edulis improves water quality through filtration feeding and provides habitat formation and coastal protection through reef formation. Like many marine calcifiers, ocean warming, and acidification are a growing threat to these organisms-impacting their morphology and function. Museum collections are useful in assessing long-term environmental impacts on organisms in a natural multi-stressor environment, where acclimation and adaptation can be considered. Using the American Museum of Natural History collections ranging from the early 1900s until now, we show that shell porosity changes through time. Shells collected today are significantly more porous than shells collected in the 1960s and, at some sites, than shells collected from the early 1900s. The disparity between porosity changes matches well with the warming that occurred over the last 130 years in the north Atlantic suggesting that warming is causing porosity changes. However, more work is required to discern local environmental impacts and to fully identify porosity drivers. Since, porosity is known to affect structural integrity, porosity increasing through time could have negative consequences for mussel reef structural integrity and hence habitat formation and storm defenses.

Crystal orientation mapping and microindentation reveal anisotropy in Porites skeletons.

Structures made by scleractinian corals support diverse ocean ecosystems. Despite the importance of coral skeletons and their predicted vulnerability to climate change, few studies have examined the mechanical and crystallographic properties of coral skeletons at the micro- and nano-scales. Here, we investigated the interplay of crystallographic and microarchitectural organization with mechanical anisotropy within Porites skeletons by measuring Young's modulus and hardness along surfaces transverse and longitudinal to the primary coral growth direction. We observed micro-scale anisotropy, where the transverse surface had greater Young's modulus and hardness by ∼ 6 GPa and 0.2 GPa, respectively. Electron backscatter diffraction (EBSD) revealed that this surface also had a higher percentage of crystals oriented with the a-axis between ± 30-60∘, relative to the longitudinal surface, and a broader grain size distribution. Within a region containing a sharp microscale gradient in Young's modulus, nanoscale indentation mapping, energy dispersive spectroscopy (EDS), EBSD, and Raman crystallography were performed. A correlative trend showed higher Young's modulus and hardness in regions with individual crystal bases (c-axis) facing upward, and in crystal fibers relative to centers of calcification. These relationships highlight the difference in mechanical properties between scales (i.e. crystals, crystal bundles, grains). Observations of crystal orientation and mechanical properties suggest that anisotropy is driven by microscale organization and crystal packing rather than intrinsic crystal anisotropy. In comparison with previous observations of nanoscale isotropy in corals, our results illustrate the role of hierarchical architecture in coral skeletons and the influence of biotic and abiotic factors on mechanical properties at different scales. STATEMENT OF SIGNIFICANCE: Coral biomineralization and the ability of corals' skeletal structure to withstand biotic and abiotic forces underpins the success of reef ecosystems. At the microscale, we show increased skeletal stiffness and hardness perpendicular to the coral growth direction. By comparing nano- and micro-scale indentation results, we also reveal an effect of hierarchical architecture on the mechanical properties of coral skeletons and hypothesize that crystal packing and orientation result in microscale anisotropy. In contrast to previous findings, we demonstrate that mechanical and crystallographic properties of coral skeletons can vary between surface planes, within surface planes, and at different analytical scales. These results improve our understanding of biomineralization and the effects of scale and direction on how biomineral structures respond to environmental stimuli.

Coral-associated nitrogen fixation rates and diazotrophic diversity on a nutrient-replete equatorial reef.

The role of diazotrophs in coral physiology and reef biogeochemistry remains poorly understood, in part because N2 fixation rates and diazotrophic community composition have only been jointly analyzed in the tissue of one tropical coral species. We performed field-based 15N2 tracer incubations during nutrient-replete conditions to measure diazotroph-derived nitrogen (DDN) assimilation into three species of scleractinian coral (Pocillopora acuta, Goniopora columna, Platygyra sinensis). Using multi-marker metabarcoding (16S rRNA, nifH, 18S rRNA), we analyzed DNA- and RNA-based communities in coral tissue and skeleton. Despite low N2 fixation rates, DDN assimilation supplied up to 6% of the holobiont's N demand. Active coral-associated diazotrophs were chiefly Cluster I (aerobes or facultative anaerobes), suggesting that oxygen may control coral-associated diazotrophy. Highest N2 fixation rates were observed in the endolithic community (0.20 µg N cm-2 per day). While the diazotrophic community was similar between the tissue and skeleton, RNA:DNA ratios indicate potential differences in relative diazotrophic activity between these compartments. In Pocillopora, DDN was found in endolithic, host, and symbiont compartments, while diazotrophic nifH sequences were only observed in the endolithic layer, suggesting a possible DDN exchange between the endolithic community and the overlying coral tissue. Our findings demonstrate that coral-associated diazotrophy is significant, even in nutrient-rich waters, and suggest that endolithic microbes are major contributors to coral nitrogen cycling on reefs.

An update of the Pb isotope inventory in post leaded-petrol Singapore environments.

Pb is a trace metal that tracks anthropogenic pollution in natural environments. Despite recent leaded petrol phase out around Southeast Asia, the region's growth has resulted in continued exposure of Pb from a variety of sources. In this study, sources of Pb into Singapore, a highly urbanised city-state situated in the central axis of Southeast Asia, are investigated using isotopic ratios and concentrations. We compiled data from our previous analyses of aerosols, incineration fly ash and sediments, with new data from analyses of soil from gas stations, water from runoff and round-island coastal seawater to obtain a spatio-temporal overview of sources of Pb into the Singapore environment. Using 206Pb/207Pb ratio, we identified three main Pb source origins: natural Pb (1.215 ± 0.001), historic/remnant leaded petrol (1.123 ± 0.013), and present-day industrial and incinerated waste (1.148 ± 0.005). Deep reservoir sediments bore larger traces of Pb from leaded petrol, but present-day runoff waters and coastal seawater were a mix of industrial and natural sources with somewhat variable concentrations. We found temporal variability in Pb isotopic ratio in aerosols indicating alternating transboundary Pb sources to Singapore that correspond to seasonal changes in monsoon winds. By contrast, seasonal monsoon circulation did not significantly influence isotopic ratios of coastal seawater Pb. Instead, seawater Pb was driven more by location differences, suggesting stronger local-scale drivers of Pb such as point sources, water flushing, and isotope exchange. The combination of multiple historic and current sources of Pb shown in this study highlights the need for continued monitoring of Pb in Southeast Asia, especially in light of emerging industries and potential large sources of Pb such as coal combustion.

Two centuries of limited variability in subtropical North Atlantic thermocline ventilation.

Ventilation and mixing of oceanic gyres is important to ocean-atmosphere heat and gas transfer, and to mid-latitude nutrient supply. The rates of mode water formation are believed to impact climate and carbon exchange between the surface and mid-depth water over decadal periods. Here, a record of (14)C/(12)C (1780-1940), which is a proxy for vertical ocean mixing, from an annually banded coral from Bermuda, shows limited inter-annual variability and a substantial Suess Effect (the decrease in (14)C/(12)C since 1900). The Sargasso Sea mixing rates between the surface and thermocline varied minimally over the past two centuries, despite changes to mean-hemispheric climate, including the Little Ice Age and variability in the North Atlantic Oscillation. This result indicates that regional formation rates of sub-tropical mode water are stable over decades, and that anthropogenic carbon absorbed by the ocean does not return to the surface at a variable rate.

Temporal distributions of anthropogenic Al, Zn and Pb in Hong Kong Porites coral during the last two centuries.

A 182-year long record of trace metal concentrations of aluminum, zinc and lead was reconstructed from a massive Porites coral skeleton from southeastern Hong Kong to evaluate the impacts of anthropogenic activity on the marine environment. Zn/Ca and Pb/Ca ratios fluctuate synchronously from the early 19th century to the present, indicating that the marine environment has been anthropogenically influenced since industrialization. Additionally, land reclamation, mining, and ship building activities are recorded by elevated Al/Ca ratios from 1900 to 1950. The coral record indicates that high levels of Zn, Pb and Al occur coincidentally with local wars, and may have contributed to partial colony mortality. Pb/Ca does not correlate well with hemispheric proxy records after 1950, indicating that coastal corals may be recording local rather than hemispheric contamination. Pb/Ca levels in Hong Kong, Guangdong and Hainan corals imply a continuous supply of Pb-based contamination to southern China not reflected in hemispheric signals.