Development of a colorimetric microfluidic pH sensor for autonomous seawater measurements

High quality carbonate chemistry measurements are required in order to fully understand the dynamics of the oceanic carbonate system. Seawater pH data with good spatial and temporal coverage are particularly critical to apprehend ocean acidification phenomena and their consequences. There is a growing need for autonomous in situ instruments that measure pH on remote platforms. Our aim is to develop an accurate and precise autonomous in situ pH sensor for long term deployment on remote platforms. The widely used spectrophotometric pH technique is capable of the required high-quality measurements. We report a key step toward the miniaturisation of a colorimetric pH sensor with the successful implementation of a simple microfluidic design with low reagent consumption. The system is particularly adapted to shipboard deployment: high quality data was obtained over a period of more than a month during a shipboard deployment in northwest European shelf waters, and less than 30 mL of indicator was consumed. The system featured a short term precision of 0.001 pH (n=20) and an accuracy within the range of a certified Tris buffer (0.004 pH). The quality of the pH system measurements have been checked using various approaches: measurements of certified Tris buffer, measurement of certified seawater for DIC and TA, comparison of measured pH against calculated pH from pCO₂, DIC and TA during the cruise in northwest European shelf waters. All showed that our measurements were of high quality. The measurements were made close to in situ temperature (+0.2 °C) in a sampling chamber which had a continuous flow of the ship’s underway seawater supply. The optical set up was robust and relatively small due to the use of an USB mini-spectrometer, a custom made polymeric flow cell and an LED light source. The use of a three wavelength LED with detection that integrated power across the whole of each LED output spectrum indicated that low wavelength resolution detectors can be used instead of the current USB mini spectrophotometer. Artefacts due to the polychromatic light source and inhomogeneity in the absorption cell are shown to have a negligible impact on the data quality. The next step in the miniaturisation of the sensor will be the incorporation of a photodiode as detector to replace the spectrophotometer.

Development and validation of an experimental life support system for assessing the effects of global climate change and environmental contamination on estuarine and coastal marine benthic communities

An experimental life support system (ELSS) was constructed to study, the interactive effects of multiple stressors on coastal and estuarine benthic communities, specifically perturbations driven by global climate change and anthropogenic environmental contamination. The ELSS allows researchers to control salinity, pH, temperature, ultraviolet radiation (UVR), tidal rhythms and exposure to selected contaminants. Unlike most microcosms previously described, our system enables true independent replication (including randomisation). In addition to this, it can be assembled using commercially available materials and equipment, thereby facilitating the replication of identical experimental setups in different geographical locations. Here, we validate the reproducibility and environmental quality of the system by comparing chemical and biological parameters recorded in our ELSS with those prevalent in the natural environment. Water, sediment microbial community and ragworm (the polychaete Hediste diversicolor) samples were obtained from four microcosms after 57 days of operation. In general, average concentrations of dissolved inorganic nutrients (NO₃^-; NH₄⁺ and PO₄ ⁻³) in the water-column of the ELSS experimental control units were within the range of concentrations recorded in the natural environment. While some shifts in bacterial community composition were observed between in situ and ELSS sediment samples, the relative abundance of most metabolically active bacterial taxa appeared to be stable. In addition, ELSS operation did not significantly affect survival, oxidative stress and neurological biomarkers of the model organism H. diversicolor. The validation data indicates that this system can be used to assess independent or interactive effects of climate change and environmental contamination on benthic communities. Researchers will be able to simulate the effects of these stressors on processes driven by microbial communities, sediment and seawater chemistry and to evaluate potential consequences to sediment toxicity using model organisms such as Hediste diversicolor.

The real limits to marine life: a further critique of the Respiration Index (update)

The recently proposed “Respiration Index” (RI = log PO₂/PCO₂) suggests that aerobic metabolism is limited by the ratio of reactants (oxygen) to products (carbon dioxide) according to the thermodynamics of cellular respiration. Here, we demonstrate further that, because of the large standard free energy change for organic carbon oxidation (ΔG° = −686 kcal mol⁻¹), carbon dioxide can never reach concentrations that would limit the thermodynamics of this reaction. A PCO₂ to PO₂ ratio of 10⁵⁰³ would be required to reach equilibrium (equilibrium constant, K_eq = 10⁵⁰³), where ΔG = 0. Thus, a Respiration Index of −503 would be the real thermodynamic limit to aerobic life. Such a Respiration Index is never reached, either in the cell or in the environment. Moreover, cellular respiration and oxygen provision are kinetically controlled such that, within limits, environmental oxygen and CO₂ concentrations have little to do with intracellular concentrations. The RI is fundamentally different from the aragonite saturation state, a thermodynamic index used to quantify the potential effect of CO₂ on calcification rates, because of its failure to incorporate the equilibrium constant of the reaction. Not only is the RI invalid, but its use leads to incorrect and misleading predictions of the threat of changing oxygen and carbon dioxide to marine life. We provide a physiological framework that identifies oxygen thresholds and allows for synergistic effects of ocean acidification and global warming.

A potential tool for high-resolution monitoring of ocean acidification

Current anthropogenic carbon dioxide emissions generate besides global warming unprecedented acidification rates of the oceans. Recent evidence indicates the possibility that ocean acidification and low oceanic pH may be a major reason for several mass extinctions in the past. However, a major bottleneck for research on ocean acidification is long-term monitoring and the collection of consistent high-resolution pH measurements. This study presents a low-power (< 1 W) small sample volume (25 μl) semiconductor based fluorescence method for real-time ship-board pH measurements at high temporal and spatial resolution (approximately 15 s and 100 m between samples). A 405 nm light emitting diode and the blue and green channels from a digital camera was used for swift detection of fluorescence from the pH sensitive dye 6,8-Dihydroxypyrene-1,3-disulfonic acid in real-time. Main principles were demonstrated by automated continuous measurements of pH in the surface water across the Baltic Sea and the Kattegat region with a large range in salinity (∼ 3–30) and temperature (∼ 0–25 °C). Ship-board precision of salinity and temperature adjusted pH measurements were estimated as low as 0.0001 pH units.

Interactive effects of global climate change and pollution on marine microbes: the way ahead

Global climate change has the potential to seriously and adversely affect marine ecosystem functioning. Numerous experimental and modeling studies have demonstrated how predicted ocean acidification and increased ultraviolet radiation (UVR) can affect marine microbes. However, researchers have largely ignored interactions between ocean acidification, increased UVR and anthropogenic pollutants in marine environments. Such interactions can alter chemical speciation and the bioavailability of several organic and inorganic pollutants with potentially deleterious effects, such as modifying microbial-mediated detoxification processes. Microbes mediate major biogeochemical cycles, providing fundamental ecosystems services such as environmental detoxification and recovery. It is, therefore, important that we understand how predicted changes to oceanic pH, UVR, and temperature will affect microbial pollutant detoxification processes in marine ecosystems. The intrinsic characteristics of microbes, such as their short generation time, small size, and functional role in biogeochemical cycles combined with recent advances in molecular techniques (e.g., metagenomics and metatranscriptomics) make microbes excellent models to evaluate the consequences of various climate change scenarios on detoxification processes in marine ecosystems. In this review, we highlight the importance of microbial microcosm experiments, coupled with high-resolution molecular biology techniques, to provide a critical experimental framework to start understanding how climate change, anthropogenic pollution, and microbiological interactions may affect marine ecosystems in the future.

Progress in controlled in situ ocean acidification experiments

Ocean acidification is widely recognized as a significant climate-related oceanic threat, not only independently but also in connection with other oceanic stressors, including warming and deoxygenation. Recent work shows that ocean acidification will negatively affect processes such as calcification of most species, including reef-building corals, and could also cause diminished fish sensory ability and respiratory stress. However, almost all of these findings result from short-term experiments on organisms in laboratory aquaria. But how can scientists perform long-term in situ experiments that may confirm, or modify, conclusions drawn from laboratory experiments? With funding from the BNP Paribas Foundation, the xFOCE workshop brought together a group of 20 scientists and engineers to examine this.

Technical Note: the determination of enclosed water volume in large flexible-wall mesocosms “KOSMOS” (update)

The volume of water enclosed inside flexible-wall mesocosm bags is hard to estimate using geometrical calculations and can be strongly variable among bags of the same dimensions. Here we present a method for precise water volume determination in mesocosms using salinity as a tracer. Knowledge of the precise volume of water enclosed allows establishment of exactly planned treatment concentrations and calculation of elemental budgets.

The mathematics of the total alkalinity–pH equation: pathway to robust and universal solution algorithms

The total alkalinity–pH equation, which relates total alkalinity and pH for a given set of total concentrations of the acid-base systems that contribute to total alkalinity in a given water sample, is reviewed and its mathematical properties established. We prove that the equation function is strictly monotone and always has exactly one positive root. Different commonly used approximations are discussed and compared. An original method to derive appropriate initial values for the iterative solution of the cubic polynomial equation based upon carbonate-borate-alkalinity is presented. We then review different methods that have been used to solve the total alkalinity–pH equation, with a main focus on biogeochemical models. The shortcomings and limitations of these methods are made out and discussed. We then present two variants of a new, robust and universally convergent algorithm to solve the total alkalinity–pH equation. This algorithm does not require any a priori knowledge of the solution. The iterative procedure is shown to converge from any starting value to the physical solution. The extra computational cost for the convergence security is only 10–15% compared to the fastest algorithm in our test series.

Technical Note: A mobile sea-going mesocosm system – new opportunities for ocean change research (update)

One of the great challenges in ocean change research is to understand and forecast the effects of environmental changes on pelagic communities and the associated impacts on biogeochemical cycling. Mesocosms, experimental enclosures designed to approximate natural conditions, and in which environmental factors can be manipulated and closely monitored, provide a powerful tool to close the gap between small-scale laboratory experiments and observational and correlative approaches applied in field surveys. Existing pelagic mesocosm systems are stationary and/or restricted to well-protected waters. To allow mesocosm experimentation in a range of hydrographic conditions and in areas considered most sensitive to ocean change, we developed a mobile sea-going mesocosm facility, the Kiel Off-Shore Mesocosms for Future Ocean Simulations (KOSMOS). The KOSMOS platform, which can be transported and deployed by mid-sized research vessels, is designed for operation in moored and free-floating mode under low to moderate wave conditions (up to 2.5 m wave heights). It encloses a water column 2 m in diameter and 15 to 25 m deep (∼50–75 m³ in volume) without disrupting the vertical structure or disturbing the enclosed plankton community. Several new developments in mesocosm design and operation were implemented to (i) minimize differences in starting conditions between mesocosms, (ii) allow for extended experimental duration, (iii) precisely determine the mesocosm volume, (iv) determine air–sea gas exchange, and (v) perform mass balance calculations. After multiple test runs in the Baltic Sea, which resulted in continuous improvement of the design and handling, the KOSMOS platform successfully completed its first full-scale experiment in the high Arctic off Svalbard (78°56.2′ N, 11°53.6′ E) in June/July 2010. The study, which was conducted in the framework of the European Project on Ocean Acidification (EPOCA), focused on the effects of ocean acidification on a natural plankton community and its impacts on biogeochemical cycling and air–sea exchange of climate-relevant gases. This manuscript describes the mesocosm hardware, its deployment and handling, CO₂ manipulation, sampling and cleaning, including some further modifications conducted based on the experiences gained during this study.

Experience with moored observations in the western Gulf of Maine from 2006 to 2012

The University of New Hampshire is studying CO₂ gas exchange, ocean acidification, air-sea dynamics, and associated biological processes in the western Gulf of Maine. Two buoys provide data supporting these studies. The UNH CO₂ buoy has been deployed jointly with the National Oceanic and Atmospheric Administration (NOAA)’s Pacific Marine Environmental Laboratory northeast of the Isles of Shoals since 2006. The Jeffreys Ledge Moored Observatory is a development mooring testing new techniques and is deployed east of Gloucester, MA. This mooring is testing the direct covariance measurement of wind stress using a 3-D sonic anemometer with a motion package to remove buoy motion effects. A fast-rate atmospheric CO₂ sensor is mounted by the anemometer to evaluate its potential for direct covariance gas flux measurements. Both buoys have additional meteorological and oceanographic sensors to provide supporting measurements. Six years of CO₂ buoy data have helped quantify the seasonal air-sea flux cycle of CO₂ in the Western Gulf of Maine. The buoy is now a node in near-term ocean carbon cycle process control experiments and longer-term ocean acidification monitoring. The Jeffreys Ledge buoy momentum flux measurements using wind and motion measurements indicate reasonable first-order buoy motion corrections can be made. Also, buoy-induced flow disturbance requires postmeasurement corrections. Rapid buoy azimuthal rotations were corrected with the addition of a steering vane. A vertical array of oxygen sensors captures phytoplankton bloom signatures and provides net community production estimates that augment in-water SAMI-CO₂ measurements and add to a robust system to support process studies and improved biophysical modeling within this region.