The effect of bacteria on planula-larvae settlement and metamorphosis in the octocoral Rhytisma fulvum fulvum

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Freire, I., Gutner-Hoch, E., Muras, A., Benayahu, Y., & Otero, A. The effect of bacteria on planula-larvae settlement and metamorphosis in the octocoral Rhytisma fulvum fulvum. Plos One, 14(9), (2019): e0223214, doi:10.1371/journal.pone.0223214.While increasing evidence supports a key role of bacteria in coral larvae settlement and development, the relative importance of environmentally-acquired versus vertically-transferred bacterial population is not clear. Here we have attempted to elucidate the role of post-brooding-acquired bacteria on the development of planula-larvae of the octocoral Rhytisma f. fulvum, in an in vitro cultivation system employing different types of filtered (FSW) and autoclaved (ASW) seawater and with the addition of native bacteria. A good development of larvae was obtained in polystyrene 6-well cell culture plates in the absence of natural reef substrata, achieving a 60–80% of larvae entering metamorphosis after 32 days, even in bacteria-free seawater, indicating that the bacteria acquired during the brooding period are sufficient to support planulae development. No significant difference in planulae attachment and development was observed when using 0.45 μm or 0.22 μm FSW, although autoclaving the 0.45 μm FSW negatively affected larval development, indicating the presence of beneficial bacteria. Autoclaving the different FSW homogenized the development of the larvae among the different treatments. The addition of bacterial strains isolated from the different FSW did not cause any significant effect on planulae development, although some specific strains of the genus Alteromonas seem to be beneficial for larvae development. Light was beneficial for planulae development after day 20, although no Symbiodinium cells could be observed, indicating either that light acts as a positive cue for larval development or the presence of beneficial phototrophic bacteria in the coral microbiome. The feasibility of obtaining advanced metamorphosed larvae in sterilized water provides an invaluable tool for studying the physiological role of the bacterial symbionts in the coral holobiont and the specificity of bacteria-coral interactions.This work was supported by: EU FP7-Research Infrastructure Initiative Assemble (Association of European marine biological laboratories); EU FP7 Project Byefouling (grant agreement no 612717); Xunta de Galicia, Consellería de Cultura, Educación e Ordenación Universitaria (grant number ED431D 2017/22). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Evidence for Rhythmicity Pacemaker in the Calcification Process of Scleractinian Coral

Reef-building scleractinian (stony) corals are among the most efficient bio-mineralizing organisms in nature. The calcification rate of scleractinian corals oscillates under ambient light conditions, with a cyclic, diurnal pattern. A fundamental question is whether this cyclic pattern is controlled by exogenous signals or by an endogenous 'biological-clock' mechanism, or both. To address this problem, we have studied calcification patterns of the Red Sea scleractinian coral Acropora eurystoma with frequent measurements of total alkalinity (AT) under different light conditions. Additionally, skeletal extension and ultra-structure of newly deposited calcium carbonate were elucidated with Sr-86 isotope labeling analysis, combined with NanoSIMS ion microprobe and scanning electron microscope imaging. Our results show that the calcification process persists with its cyclic pattern under constant light conditions while dissolution takes place within one day of constant dark conditions, indicating that an intrinsic, light-entrained mechanism may be involved in controlling the calcification process in photosymbiotic corals

A unique coral biomineralization pattern has resisted 40 million years of major ocean chemistry change

Today coral reefs are threatened by changes to seawater conditions associated with rapid anthropogenic global climate change. Yet, since the Cenozoic, these organisms have experienced major fluctuations in atmospheric CO2 levels (from greenhouse conditions of high pCO2 in the Eocene to low pCO2 ice-house conditions in the Oligocene-Miocene) and a dramatically changing ocean Mg/Ca ratio. Here we show that the most diverse, widespread, and abundant reef-building coral genus Acropora (20 morphological groups and 150 living species) has not only survived these environmental changes, but has maintained its distinct skeletal biomineralization pattern for at least 40 My: Well-preserved fossil Acropora skeletons from the Eocene, Oligocene, and Miocene show ultra-structures indistinguishable from those of extant representatives of the genus and their aragonitic skeleton Mg/Ca ratios trace the inferred ocean Mg/Ca ratio precisely since the Eocene. Therefore, among marine biogenic carbonate fossils, well-preserved acroporid skeletons represent material with very high potential for reconstruction of ancient ocean chemistry

The role of skeletal micro-architecture in diagenesis and dating of Acropora palmata

Past variations in global sea-level reflect continental ice volume, a crucial factor for understanding the Earth's climate system. The Caribbean coral Acropora palmata typically forms dense stands in very shallow water and therefore fossil samples mark past sea-level. Uranium-series methods are commonly used to establish a chronology for fossil coral reefs, but are compromised by post mortem diagenetic changes to coral skeleton. Current screening approaches are unable to identify all altered samples, whilst models that attempt to correct for 'open-system' behaviour are not applicable across all diagenetic scenarios. In order to better understand how U-series geochemistry varies spatially with respect to diagenetic textures, we examine these aspects in relation to skeletal micro-structure and intra-crystalline amino acids, comparing an unaltered modern coral with a fossil A. palmata colony containing zones of diagenetic alteration (secondary overgrowth of aragonite, calcite cement and dissolution features). We demonstrate that the process of skeletogenesis in A. palmata causes heterogeneity in porosity, which can account for the observed spatial distribution of diagenetic features; this in turn explains the spatially-systematic trends in U-series geochemistry and consequently, U-series age. We propose a scenario that emphasises the importance of through-flow of meteoric waters, invoking both U-loss and absorption of mobilised U and Th daughter isotopes. We recommend selective sampling of low porosity A. palmata skeleton to obtain the most reliable U-series ages. We demonstrate that intra-crystalline amino acid racemisation (AAR) can be applied as a relative dating tool in Pleistocene A. palmata samples that have suffered heavy dissolution and are therefore unsuitable for U-series analyses. Based on relatively high intra-crystalline concentrations and appropriate racemisation rates, glutamic acid and valine are most suited to dating mid-late Pleistocene A. palmata. Significantly, the best-preserved material in the fossil specimen yields a U-series age of 165 ± 8 ka, recording a paleo sea-level of -35 ± 7 msl during the MIS 6.5 interstadial on Barbados

Simultaneous extension of both basic microstructural components in scleractinian coral skeleton during night and daytime, visualized by in situ Sr-86 pulse labeling

Using in situ (12 h) pulse-labeling of scleractinian coral aragonitic skeleton with stable 86Sr isotope, the diel pattern of skeletal extension was investigated in the massive Porites lobata species, grown at 5 m depth in the Gulf of Eilat Several microstructural aspects of coral biomineralization were elucidated, among which the most significant is simultaneous extension of the two basic microstructural components Rapid Accretion Deposits (RAD; also called Centers of Calcification) and Thickening Deposits (TD; also called fibers), both at night and during daytime. Increased thickness of the 86Sr-labeled growth-front in the RADs compared to the adjacent TDs revealed that in this species RADs extend on average twice as fast as TDs. At the level of the individual corallite, skeletal extension is spatially highly heterogeneous, with sporadic slowing or cessation depending on growth directions and skeletal structure morphology. Daytime photosynthesis by symbiotic dinoflagellates is widely acknowledged to substantially increase calcification rates at the colony and the corallite level in reef-building corals. However, in our study, the average night-time extension rate (visualized in three successive 12 h pulses) was similar to the average daytime extension (visualized in the initial 12 h pulse), in all growth directions and skeletal structures. This research provides a platform for further investigations into the temporal calibration of coral skeletal extension via cyclic growth increment deposition, which is a hallmark of coral biomineralization

Simultaneous extension of both basic microstructural components in scleractinian coral skeleton during night and daytime, visualized by in situ ⁸⁶Sr pulse labeling

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Antimacrofouling Efficacy of Innovative Inorganic Nanomaterials Loaded with Booster Biocides

The application of nano-structured compounds has been increasing rapidly in recent years, in several fields. The use of engineered nano-materials as carriers of antifouling compounds is just beginning and already reveals clear advantages compared to bulk active compounds, such as slowed and controlled release, novel functionality, and high loading capacity. This present study assesses the antifouling efficacy of two nanostructured materials, spherical mesoporous silica nanocapsules (SiNC) and Zn-Al layered double hydroxides (LDH), loaded with two commercial biocides, zinc prithione (ZnPT) and copper pyrithione (CuPT). The study used adult mussels from three geographical regions, the Atlantic Ocean, Mediterranean Sea, and the Red Sea, to examine the efficacy of the innovative compounds. The efficacy of these compounds on larvae of the bryozoan Bugula neritina from the Mediterranean Sea and the Red Sea was also examined. The results of this study demonstrated the environmentally friendly properties of unloaded LDH against the two-model systems, adult mussels or bryozoan larvae. ZnPT entrapped in LDH demonstrated the most effective antifouling compound against the two model systems. A comparison of the impact of the two compounds on macrofouling organisms from the different marine habitats examined in this study indicates a distinction associated with the organisms’ different ecosystems. The Red Sea mussels and bryozoans, representing a tropical marine ecosystem, yielded the highest efficacy values among tested Atlantic Ocean and Mediterranean Sea mussels and bryozoans