Sea butterflies in a pickle: reliable biomarkers and seasonal sensitivity of Limacina retroversa to ocean acidification in the Gulf of Maine.

The passive dissolution of anthropogenically produced CO2 into the ocean system is reducing ocean pH and changing a suite of chemical equilibria, with negative consequences for some marine organisms, in particular those that bear calcium carbonate shells. Although our monitoring of these chemical changes has improved, we have not developed effective tools to translate observations, which are typically of the pH and carbonate saturation state, into ecologically relevant predictions of biological risks. One potential solution is to develop bioindicators: biological variables with a clear relationship to environmental risk factors that can be used for assessment and management. Thecosomatous pteropods are a group of pelagic shelled marine gastropods, whose biological responses to CO2 have been suggested as potential bioindicators of ocean acidification owing to their sensitivity to acidification in both the laboratory and the natural environment. Using five CO2 exposure experiments, occurring across four seasons and running for up to 15 days, we describe a consistent relationship between saturation state, shell transparency and duration of exposure, as well as identify a suite of genes that could be used for biological monitoring with further study. We clarify variations in thecosome responses due to seasonality, resolving prior uncertainties and demonstrating the range of their phenotypic plasticity. These biomarkers of acidification stress can be implemented into ecosystem models and monitoring programmes in regions where pteropods are found, whilst the approach will serve as an example for other regions on how to bridge the gap between point-based chemical monitoring and biologically relevant assessments of ecosystem health.

Development of the Channelized Optical System II for In Situ, High-Frequency Measurements of Dissolved Inorganic Carbon in Seawater.

This study describes the development of the CHANnelized Optical System II (CHANOS II), an autonomous, in situ sensor capable of measuring seawater dissolved inorganic carbon (DIC) at high frequency (up to ∼1 Hz). In this sensor, CO2 from acidified seawater is dynamically equilibrated with a pH-sensitive indicator dye encapsulated in gas-permeable Teflon AF 2400 tubing. The pH in the CO2 equilibrated indicator is measured spectrophotometrically and can be quantitatively correlated to the sample DIC. Ground-truthed field data demonstrate the sensor's capabilities in both time-series measurements and surface mapping in two coastal sites across tidal cycles. CHANOS II achieved an accuracy and precision of ±5.9 and ±5.5 µmol kg-1. The mean difference between traditional bottle and sensor measurements was -3.7 ± 10.0 (1σ) µmol kg-1. The sensor can perform calibration in situ using Certified Reference Materials (CRMs) to ensure measurement quality. The coastal time-series measurements highlight high-frequency variability and episodic biogeochemical shifts that are difficult to capture by traditional methods. Surface DIC mapping shows multiple endmembers in an estuary and highlights fine-scale spatial variabilities of DIC. The development of CHANOS II demonstrates a significant technological advance in seawater CO2 system sensing, which enables high-resolution, subsurface time-series, and profiling deployments.

Carbonate chemistry and carbon sequestration driven by inorganic carbon outwelling from mangroves and saltmarshes.

Mangroves and saltmarshes are biogeochemical hotspots storing carbon in sediments and in the ocean following lateral carbon export (outwelling). Coastal seawater pH is modified by both uptake of anthropogenic carbon dioxide and natural biogeochemical processes, e.g., wetland inputs. Here, we investigate how mangroves and saltmarshes influence coastal carbonate chemistry and quantify the contribution of alkalinity and dissolved inorganic carbon (DIC) outwelling to blue carbon budgets. Observations from 45 mangroves and 16 saltmarshes worldwide revealed that >70% of intertidal wetlands export more DIC than alkalinity, potentially decreasing the pH of coastal waters. Porewater-derived DIC outwelling (81 ± 47 mmol m-2 d-1 in mangroves and 57 ± 104 mmol m-2 d-1 in saltmarshes) was the major term in blue carbon budgets. However, substantial amounts of fixed carbon remain unaccounted for. Concurrently, alkalinity outwelling was similar or higher than sediment carbon burial and is therefore a significant but often overlooked carbon sequestration mechanism.

Dynamic energy budget modeling of Atlantic surfclam, Spisula solidissima, under future ocean acidification and warming.

A dynamic energy budget (DEB) model integrating pCO2 was used to describe ocean acidification (OA) effects on Atlantic surfclam, Spisula solidissima, bioenergetics. Effects of elevated pCO2 on ingestion and somatic maintenance costs were simulated, validated, and adapted in the DEB model based upon growth and biological rates acquired during a 12-week laboratory experiment. Temperature and pCO2 were projected for the next 100 years following the intergovernmental panel on climate change representative concentration pathways scenarios (2.6, 6.0, and 8.5) and used as forcing variables to project surfclam growth and reproduction. End-of-century water warming and acidification conditions resulted in simulated faster growth for young surfclams and more energy allocated to reproduction until the beginning of the 22nd century when a reduction in maximum shell length and energy allocated to reproduction was observed for the RCP 8.5 scenario.

In situ sensor technology for simultaneous spectrophotometric measurements of seawater total dissolved inorganic carbon and pH.

A new, in situ sensing system, Channelized Optical System (CHANOS), was recently developed to make high-resolution, simultaneous measurements of total dissolved inorganic carbon (DIC) and pH in seawater. Measurements made by this single, compact sensor can fully characterize the marine carbonate system. The system has a modular design to accommodate two independent, but similar measurement channels for DIC and pH. Both are based on spectrophotometric detection of hydrogen ion concentrations. The pH channel uses a flow-through, sample-indicator mixing design to achieve near instantaneous measurements. The DIC channel adapts a recently developed spectrophotometric method to achieve flow-through CO2 equilibration between an acidified sample and an indicator solution with a response time of only ∼ 90 s. During laboratory and in situ testing, CHANOS achieved a precision of ±0.0010 and ± 2.5 µmol kg(-1) for pH and DIC, respectively. In situ comparison tests indicated that the accuracies of the pH and DIC channels over a three-week time-series deployment were ± 0.0024 and ± 4.1 µmol kg(-1), respectively. This study demonstrates that CHANOS can make in situ, climatology-quality measurements by measuring two desirable CO2 parameters, and is capable of resolving the CO2 system in dynamic marine environments.

Automated spectrophotometric analyzer for rapid single-point titration of seawater total alkalinity.

An automated analyzer was developed to achieve fast, precise, and accurate measurements of seawater total alkalinity (AT) based on single-point titration and spectrophotometric pH detection. The single-point titration was carried out in a circulating loop, which allowed the titrant (hydrochloric acid and bromocresol green solution) and a seawater sample to mix at a constant volume ratio. The dissolved CO2 in the sample-titrant mixture was efficiently removed by an inline CO2 remover, which consists of a gas-permeable tubing (Teflon AF2400) submerged in a sodium hydroxide (NaOH) solution. The pH of the mixture was then measured with a custom-made spectrophotometric detection system. The analyzer was calibrated against multiple certified reference materials (CRMs) with different AT values. The analyzer features a sample throughput time of 6.5 min with high precision (±0.33-0.36 µmol kg(-1); n = 48) and accuracy (-0.33 ± 0.99 µmol kg(-1); n = 10). Intercomparison to a traditional open-cell AT titrator showed overall good agreement of 0.88 ± 2.03 µmol kg(-1) (n = 22). The analyzer achieved excellent stability without recalibration over 11 days, during which time 320 measurements were made with a total running time of over 40 h. Because of its small size, low power consumption requirements, and its ability to be automated, the new analyzer can be adapted for underway and in situ measurements.

High-frequency spectrophotometric measurements of total dissolved inorganic carbon in seawater.

A new spectrophotometric method was developed to achieve continuous measurements of total dissolved inorganic carbon (DIC) in seawater. It uses a countercurrent flow design and a highly CO2-permeable membrane (Teflon AF 2400) to achieve flow-through CO2 equilibration between an acidified sample and an indicator solution with a fast response time of ~22 s. This method improves the spatiotemporal resolution by more than 1 order of magnitude compared to the existing spectrophotometric method. The flow-through equilibration allows for continuous (~1 Hz) detection and real-time data smoothing. The method had a short-term precision of ± 2.0 µmol kg(-1) for a given flow-through sample. It achieved a field precision of ± 3.6 µmol kg(-1) and successfully captured high DIC variability down to minute scales. Measurements by the new method over the typical range of oceanic DIC showed good agreement with measurements made by an established method (mean differences -1.6 to 0.3 µmol kg(-1) with 1σ ± 6.0-6.7 µmol kg(-1)). This level of precision and accuracy is comparable to that of the existing spectrophotometric method. The characteristics of the new method make it particularly suitable for high-frequency, submerged measurements required for mobile observing platforms in the ocean. It can also be adapted for high-frequency, spectrophotometric measurements of seawater CO2 fugacity.

Simultaneous spectrophotometric flow-through measurements of pH, carbon dioxide fugacity, and total inorganic carbon in seawater.

An autonomous multi-parameter flow-through CO2 system has been developed to simultaneously measure surface seawater pH, carbon dioxide fugacity (fCO2), and total dissolved inorganic carbon (DIC). All three measurements are based on spectrophotometric determinations of solution pH at multiple wavelengths using sulfonephthalein indicators. The pH optical cell is machined from a PEEK polymer rod bearing a bore-hole with an optical pathlength of approximately 15 cm. The fCO2 optical cell consists of Teflon AF 2400 (DuPont) capillary tubing sealed within the bore-hole of a PEEK rod. This Teflon AF tubing is filled with a standard indicator solution with a fixed total alkalinity, and forms a liquid core waveguide (LCW). The LCW functions as both a long pathlength (approximately 15 cm) optical cell and a membrane that equilibrates the internal standard solution with external seawater. fCO2 is then determined by measuring the pH of the internal solution. DIC is measured by determining the pH of standard internal solutions in equilibrium with seawater that has been acidified to convert all forms of DIC to CO2. The system runs repetitive measurement cycles with a sampling frequency of approximately 7 samples (21 measurements) per hour. The system was used for underway measurements of sea surface pH, fCO2, and DIC during the CLIVAR/CO2 A16S cruise in the South Atlantic Ocean in 2005. The field precisions were evaluated to be 0.0008 units for pH, 0.9 microatm for fCO2, and 2.4 micromol kg(-1) for DIC. These field precisions are close to those obtained in the laboratory. Direct comparison of our measurements and measurements obtained using established standard methods revealed that the system achieved field agreements of 0.0012+/-0.0042 units for pH, 1.0+/-2.5 microatm for fCO2, and 2.2+/-6.0 micromol kg(-1) for DIC. This system integrates spectrophotometric measurements of multiple CO2 parameters into a single package suitable for observations of both seawater and freshwater.

Spectrophotometric measurements of pH in-situ: laboratory and field evaluations of instrumental performance.

Automated in-situ instrumentation has been developed for precise and accurate measurements of a variety of analytes in natural waters. In this work we describe the use of 'SEAS' (spectrophotometric elemental analysis system) instrumentation for measurements of solution pH. SEAS-pH incorporates a CCD-based spectrophotometer, an incandescent light source, and dual pumps for mixing natural water samples with a sulfonephthalein indicator. The SEAS-pH optical cell consists of either a liquid core waveguide (LCW, Teflon AF-2400) or a custom-made PEEK cell. Long optical path lengths allow use of indicators at low concentrations, thereby precluding large indicator-induced pH perturbations. Laboratory experiments show that pH measurements obtained using LCW and PEEK optical cells are indistinguishable from measurements obtained using conventional spectrophotometric cells and high-performance spectrophotometers. Deployments in the Equatorial Pacific and the Gulf of Mexico demonstrate that SEAS-pH instruments are capable of obtaining vertical pH profiles with high spatial resolution. SEAS-pH deployments at a fixed river-site (Hillsborough River, FL) demonstrate the capability of SEAS for observations of diel pH cycles with high temporal resolution. The in-situ precision of SEAS-pH is assessed as 0.0014 pH units, and the system's measurement frequency is approximately 0.5 Hz. This work indicates that in-situ instrumentation can be used to provide accurate, precise, and highly resolved observations of carbon-system transformations in the natural environment.

A long pathlength spectrophotometric pCO(2) sensor using a gas-permeable liquid-core waveguide.

The first long pathlength fiber optic-based sensor system to measure pCO(2) in natural waters and the atmosphere is described. The sensor is based on a liquid-core (an indicator-HCO(3)(-)/CO(3)(2-) buffer solution) waveguide made of a low refractive index amorphous fluoropolymer tubing, the wall of which serves as a gas-permeable membrane to sense pCO(2) changes. The system detects the indicator absorbance changes when the liquid-core reaches CO(2) equilibrium with the surrounding sample. Theoretical calculations demonstrate that due to indicator buffer effects, increasing the optical pathlength is a more efficient way to obtain higher sensitivity than increasing the indicator concentration. Using an 18-cm cell with low indicator concentrations (10 muM), this system achieves a precision and an accuracy of +/-2-3 muatm in the pCO(2) range of 200-500 muatm. The sensor also features a response time (99%) of only 2 min for low-level (<1000 muatm) pCO(2) measurements as a result of high CO(2) permeability of the amorphous fluoropolymer membrane. Field tests indicate that this new sensor is capable of handling both atmospheric and aquatic pCO(2) monitoring.