Bacterial behavior on coated porous titanium substrates for biomedical applications

In this work, bacterial behavior on dense and porous titanium substrates is discussed. Porous titanium was fabricated by a space holder technique (using 50 vol%, NH4HCO3 with particle sizes between 250 and 355 μm). These substrates were coated by sulfonated PEEK (termed SPEEK). Characterization of the porous substrate was carried out using the Archimedes Method, Image Analysis, and three-dimensional X-ray Micro-Computed Tomography (including total and interconnected porosity, equivalent diameter, and pore shape factor), as well as mechanical characterization (specifically stiffness and yield strength). A detailed study was performed here to investigate the influence of substrate porosity on the adhesion and proliferation of E. coli, MRSA, and P. aeruginosa (common causes of orthopedic device-associated infections). Bacterial colonization was examined in terms of the initial bacterial concentration, as well as bacterial adherence to and growth on the surface and inside the pores. Results suggest that fully dense titanium supported the least bacterial colonization, while the porous titanium promoted bacterial growth in the medium and inside the cavities. Furthermore, the SPEEK coating deposited onto the samples inhibited bacteria growth inside the porous materials. In this manner, this study showed for the first time that SPEEK could have potential antibacterial properties to offset the increase in bacteria growth commonly observed in porous materials.Junta de Andalucía-FEDER (Spain) Ref. P12-TEP-1401Ministry of Economy and Competitiveness of Spain, MAT2015-71284-

Temperature Induced Physiological Reaction Norms of the Coccolithophore Gephyrocapsa oceanica and Resulting Coccolith Sr/Ca and Mg/Ca Ratios

Coccolithophores are one of the major contributors to the pelagic production of calcium carbonate and their fossilized remains are a key component of the biogeochemical cycles of calcium (Ca), magnesium (Mg), and other divalent cations present in the intracellular precipitated calcitic structures (coccoliths). The geochemical signature of coccoliths (e.g., Sr/Ca and Mg/Ca ratios) is used as paleoproxy to reconstruct past environmental conditions and to understand the underlying physiological precipitation kinetics. Here, we present the elemental fractionation of Sr and Mg in calcite of the coccolithophore Gephyrocapsa oceanica from controlled laboratory experiments applying an extended temperature gradient (12 to 27°C). The physiological reaction norm of G. oceanica, in terms of growth rate, exhibited optimum behavior while the partition coefficient of Sr (DSr) was linearly correlated with temperature and DMg indicated no specific trend. Our results indicate: (1) a presumably secondary physiological control of DSr, and (2) the importance of calibrating coccolithophore-based proxies using experiments that include the full physiological reaction norms (i.e., a possible non-linear response) to environmental drivers (e.g., temperature, salinity, and pH, etc.). The presented results contribute to an improved understanding of the underlying physiological kinetics involved in regulating coccolith elemental fractionation and give additional implications for designing future laboratory experiments to calibrate and apply coccolithophore based paleoproxies on the fossil sediment record

Balancing porosity and mechanical properties of titanium samples to favor cellular growth against bacteria

Two main problems limit the success of titanium implants: bacterial infection, which restricts their osseointegration capacity; and the stiffness mismatch between the implant and the host cortical bone, which promotes bone resorption and risk of fracture. Porosity incorporation may reduce this difference in stiffness but compromise biomechanical behavior. In this work, the relationship between the microstructure (content, size, and shape of pores) and the antibacterial and cellular behavior of samples fabricated by the space-holder technique (50 vol % NH4HCO3 and three ranges of particle sizes) is established. Results are discussed in terms of the best biomechanical properties and biofunctional activity balance (cell biocompatibility and antibacterial behavior). All substrates achieved suitable cell biocompatibility of premioblast and osteoblast in adhesion and proliferation processes. It is worth to highlighting that samples fabricated with the 100–200 μm space-holder present better mechanical behavior—in terms of stiffness, microhardness, and yield strength—which make them a very suitable material to replace cortical bone tissues. Those results exposed the relationship between the surface properties and the race of bacteria and mammalian cells for the surface with the aim to promote cellular growth over bacteria.University of Seville (Spain) VI Plan Propio de Investigación y Transferencia—US 2018, I.3A

Physiological responses of coccolithophores to abrupt exposure of naturally low pH deep seawater

Upwelling is the process by which deep, cold, relatively high-CO2, nutrient-rich seawater rises to the sunlit surface of the ocean. This seasonal process has fueled geoengineering initiatives to fertilize the surface ocean with deep seawater to enhance productivity and thus promote the drawdown of CO2. Coccolithophores, which inhabit many upwelling regions naturally ‘fertilized’ by deep seawater, have been investigated in the laboratory in the context of ocean acidification to determine the extent to which nutrients and CO2 impact their physiology, but few data exist in the field except from mesocosms. Here, we used the Porcupine Abyssal Plain (north Atlantic Ocean) Observatory to retrieve seawater from depths with elevated CO2 and nutrients, mimicking geoengineering approaches. We tested the effects of abrupt natural deep seawater fertilization on the physiology and biogeochemistry of two strains of Emiliania huxleyi of known physiology. None of the strains tested underwent cell divisions when incubated in waters obtained from 1,000 m, artificial upwelling using shallower waters may not be a suitable approach for promoting carbon sequestration for some locations and assemblages, and should therefore be investigated on a site-by-site basis

Sinking Jelly-Carbon Unveils Potential Environmental Variability along a Continental Margin

Particulate matter export fuels benthic ecosystems in continental margins and the deep sea, removing carbon from the upper ocean. Gelatinous zooplankton biomass provides a fast carbon vector that has been poorly studied. Observational data of a large-scale benthic trawling survey from 1994 to 2005 provided a unique opportunity to quantify jelly-carbon along an entire continental margin in the Mediterranean Sea and to assess potential links with biological and physical variables. Biomass depositions were sampled in shelves, slopes and canyons with peaks above 1000 carcasses per trawl, translating to standing stock values between 0.3 and 1.4 mg C m2 after trawling and integrating between 30,000 and 175,000 m2 of seabed. The benthopelagic jelly-carbon spatial distribution from the shelf to the canyons may be explained by atmospheric forcing related with NAO events and dense shelf water cascading, which are both known from the open Mediterranean. Over the decadal scale, we show that the jelly-carbon depositions temporal variability paralleled hydroclimate modifications, and that the enhanced jelly-carbon deposits are connected to a temperature-driven system where chlorophyll plays a minor role. Our results highlight the importance of gelatinous groups as indicators of large-scale ecosystem change, where jelly-carbon depositions play an important role in carbon and energy transport to benthic systems

Phenotypic Variability in the Coccolithophore Emiliania huxleyi

Coccolithophores are a vital part of oceanic phytoplankton assemblages that produce organic matter and calcium carbonate (CaCO3) containing traces of other elements (i.e. Sr and Mg). Their associated carbon export from the euphotic zone to the oceans' interior plays a crucial role in CO2 feedback mechanisms and biogeochemical cycles. The coccolithophore Emiliania huxleyi has been widely studied as a model organism to understand physiological, biogeochemical, and ecological processes in marine sciences. Here, we show the inter-strain variability in physiological and biogeochemical traits in 13 strains of E. huxleyi from various biogeographical provinces obtained from culture collections commonly used in the literature. Our results demonstrate that inter-strain genetic variability has greater potential to induce larger phenotypic differences than the phenotypic plasticity of single strains cultured under a broad range of variable environmental conditions. The range of variation found in physiological parameters and calcite Sr:Ca highlights the need to reconsider phenotypic variability in paleoproxy calibrations and model parameterizations to adequately translate findings from single strain laboratory experiments to the real ocean

Jelly biomass sinking speed reveals a fast carbon export mechanism

Sinking of gelatinous zooplankton biomass is an important component of the biological pump removing carbon from the upper ocean. The export efficiency, e.g., how much biomass reaches the ocean interior sequestering carbon, is poorly known because of the absence of reliable sinking speed data. We measured sinking rates of gelatinous particulate organic matter (jelly-POM) from different species of scyphozoans, ctenophores, thaliaceans, and pteropods, both in the field and in the laboratory in vertical columns filled with seawater using high-quality video. Using these data, we determined taxon-specific jelly-POM export efficiencies using equations that integrate biomass decay rate, seawater temperature, and sinking speed. Two depth scenarios in several environments were considered, with jelly-POM sinking from 200 and 600 m in temperate, tropical, and polar regions. Jelly-POM sank on average between 850 and 1500 m d−1 (salps: 800–1200 m d−1; ctenophores: 1200–1500 m d−1; scyphozoans: 1000–1100 m d−1; pyrosomes: 1300 m d−1). High latitudes represent a fast-sinking and low-remineralization corridor, regardless of species. In tropical and temperate regions, significant decomposition takes place above 1500 m unless jelly-POM sinks below the permanent thermocline. Sinking jelly-POM sequesters carbon to the deep ocean faster than anticipated, and should be incorporated into biogeochemical and modeling studies to provide more realistic quantification of export via the biological carbon pump worldwide

Phenotypic variability in the coccolithophore Emiliania huxleyi

Coccolithophores are a vital part of oceanic phytoplankton assemblages that produce organic matter and calcium carbonate (CaCO3) containing traces of other elements (i.e. Sr and Mg). Their associated carbon export from the euphotic zone to the oceans' interior plays a crucial role in CO2 feedback mechanisms and biogeochemical cycles. The coccolithophore Emiliania huxleyi has been widely studied as a model organism to understand physiological, biogeochemical, and ecological processes in marine sciences. Here, we show the inter-strain variability in physiological and biogeochemical traits in 13 strains of E. huxleyi from various biogeographical provinces obtained from culture collections commonly used in the literature. Our results demonstrate that inter-strain genetic variability has greater potential to induce larger phenotypic differences than the phenotypic plasticity of single strains cultured under a broad range of variable environmental conditions. The range of variation found in physiological parameters and calcite Sr:Ca highlights the need to reconsider phenotypic variability in paleoproxy calibrations and model parameterizations to adequately translate findings from single strain laboratory experiments to the real ocean

The effect of nitrate and phosphate availability on Emiliania huxleyi (NZEH) physiology under different CO2 scenarios

Growth and calcification of the marine coccolithophorid Emiliania huxleyi is affected by ocean acidification and macronutrients limitation and its response varies between strains. Here we investigated the physiological performance of a highly calcified E. huxleyi strain, NZEH, in a multiparametric experiment. Cells were exposed to different CO2 levels (ranging from 250 to 1314 μatm) under three nutrient conditions nutrient replete (R), nitrate limited (-N), and phosphate limited (-P). We focused on calcite and organic carbon quotas and on nitrate and phosphate utilization by analyzing the activity of nitrate reductase (NRase) and alkaline phosphatase (APase), respectively. Particulate inorganic (PIC) and organic (POC) carbon quotas increased with increasing CO2 under R conditions but a different pattern was observed under nutrient limitation. The PIC:POC ratio decreased with increasing CO2 in nutrient limited cultures. Coccolith length increased with CO2 under all nutrient conditions but the coccosphere volume varied depending on the nutrient treatment. Maximum APase activity was found at 561 μatm of CO2 (pH 7.92) in -P cultures and in R conditions, NRase activity increased linearly with CO2. These results suggest that E. huxleyi's competitive ability for nutrient uptake might be altered in future high-CO2 oceans. The combined dataset will be useful in model parameterizations of the carbon cycle and ocean acidification

Benthic marine calcifiers coexist with CaCO3-undersaturated seawater worldwide

Ocean acidification and decreasing seawater saturation state with respect to calcium carbonate (CaCO3) minerals have raised concerns about the consequences to marine organisms, especially those building structures made of CaCO3. A large proportion of benthic marine calcifiers incorporate Mg2+ into their calcareous structures (i.e., Mg-calcite) which, in general, reduces mineral stability. The vulnerability of some marine calcifiers to ocean acidification is related to the solubility of their calcareous structures, but not all marine organisms conform to this because of sophisticated biological and physiological mechanisms to construct and maintain CaCO3 structures. Few studies have considered seawater saturation state with respect to species-specific mineralogy in evaluating the effect of ocean acidification on marine organisms. Here, a global dataset of skeletal mol % MgCO3 of benthic calcifiers and in situ environmental conditions (temperature, salinity, pressure, and [CO32-]) spanning a depth range of 0 m (subtidal/neritic) to 5500 m (abyssal) was assembled to calculate in situ seawater saturation states with respect to species-specific Mg-calcite mineral compositions (?Mg-x). Up to 20% of all studied calcifiers at depths 1200 m currently experience seawater mineral undersaturation with respect to their skeletal mineral phase (?Mg-x1200 m) of all studied calcifying species to seawater undersaturation. These observations underscore concerns over the ability of marine benthic calcifiers to continue to construct and maintain their calcareous structures under these conditions. We advocate that ocean acidification tipping points can only be understood by assessing species-specific responses, and because of different seawater ?Mg-x present in all marine ecosystems