CO2 × temperature specific early life survival and growth of Menidia menidia assessed by 5 factorial experiments

Website: https://www.bco-dmo.org/dataset/732818
Data Type: Other Field Results, experimental
Version: 05 April 2018
Version Date: 2018-04-05

Project
» Collaborative research: Understanding the effects of acidification and hypoxia within and across generations in a coastal marine fish (HYPOA)
ContributorsAffiliationRole
Baumann, HannesUniversity of Connecticut (UConn)Principal Investigator, Contact
Murray, Christopher S.University of Connecticut (UConn - Avery Point)Co-Principal Investigator
Nye, JanetStony Brook University - SoMAS (SUNY-SB SoMAS)Co-Principal Investigator
Rauch, ShannonWoods Hole Oceanographic Institution (WHOI BCO-DMO)BCO-DMO Data Manager

Abstract
In five individual rearing experiments, wild-caught M. menidia adults were spawned to test offspring sensitivity to factorial combinations of pCO2 (nominal: 400, 2200, 4000, and 6000 µatm) and temperature (17, 20, 24, and 28 °C) through measurements of early-life survival and growth. For experiment 1, adults were collected from Poquot Beach (40.947376, -73.10258), and the experiment took place at Stony Brook University’s Flax Pond Marine Laboratory. For experiments 2–5, spawning adults were collected from Mumford Cove (41.321526, -72.015247), and experiments were conducted in the Rankin Seawater Facility at University of Connecticut’s Avery Point campus. The experiments quantified two survival and two growth traits for each replicate and CO2 × temperature treatment; embryo survival (fertilization to 1 dph), larval survival (1 dph to experiment termination), size (SL) at hatch (1 dph), and growth rate ((SL at end of experiment – SL 1dph)/number days reared post hatch). These data are published in: Murray, C.S., and Baumann, H. (2018) You Better Repeat It: Complex CO2 × Temperature Effects in Atlantic Silverside Offspring Revealed by Serial experimentation. Diversity. doi:10.3390/d10030069.


Coverage

Spatial Extent: N:41.321526 E:-72.015247 S:40.947376 W:-73.10258
Temporal Extent: 2014-05-05 - 2017-06-01

Dataset Description

CO2 x temperature specific early life survival and growth assessed by 5 factorial experiments.

These data are published in: Murray, C.S. & Baumann, H. 2018 You better repeat it: complex temperature × CO2 effects in Atlantic silverside offspring revealed by serial experimentation. Diversity 10, 1-19. doi:10.3390/d10030069


Acquisition Description

CO2 × temperature manipulations and measurements:
For 2 × 2 and 3 × 2 factorial designs, replicate rearing containers (20 L) were placed into large temperature-controlled water baths. Elevated CO2 levels were achieved via gas proportioners (ColeParmer®) mixing air with 100% CO2 (bone dry grade) that was delivered continuously to the bottom of each replicate rearing container via airstone. To counteract metabolic CO2 accumulation, control CO2 conditions were achieved by forcing compressed laboratory air through a series of CO2 stripping units containing granular soda lime (AirGas®), a particle filter (1 µm), and then to each replicate via airstone. Target pH levels were monitored daily using a handheld pH probes (Orion Ross Ultra pH/ATC Triode with Orion Star A121 pH Portable Meter; Intellical PHC281 pH Electrode with Hach® HQ11D Handheld pH/ORP Meter) calibrated bi-weekly with 2-point pHNBS references. Continuous bubbling maintained dissolved oxygen saturation (>8 mg/ DO) in rearing vessels. Target treatment temperatures were controlled by thermostats (Aqualogic®) which powered chillers (DeltaStar®) or glass submersible heaters to maintain water bath temperatures. For 3 × 3 factorial experiments, we developed an automated acidification system composed of nine discrete recirculation units designed for larval fish rearing. We designed a LabView (National Instruments®) based program to fully automate the control of seawater chemistry. The software interfaces with the recirculating units via a data-acquisition module (NI cDAQ-9184, National Instruments®), which controls nine sampling pumps (one per tank) and a series of gas and water solenoid valves, while receiving input from a central pH electrode (Hach pHD® digital electrode calibrated weekly using 2-point pHNBS references) and dissolved oxygen probe (Hach LDO® Model 2). The software sequentially assesses the pH conditions in each rearing unit (each tank once per hour) by pumping water for ~450 seconds through the housing of the central pH probe, comparing measured pH levels to set-points and then adjusting levels by bubbling standardized amounts 100% CO2 (bone dry grade, AirGas®) or CO2-stripped air into the sump of each tank. The software also maintains DO saturation (>8 mg/l) by bubbling in CO2-stripped air. LabView logs current pH, temperature, and DO conditions before cycling to the next unit. Temperatures were controlled by thermostats (Aqualogic®) that powered submersible heaters or in-line chillers (DeltaStar®).

Actual treatment CO2 levels were determined based on measurements of pH, temperature, salinity, and total alkalinity (AT). Treatment tanks were sampled three times per experiment for measurements of AT (μmol kg-1). Seawater was siphoned and filtered (to 10 µm) into 300 ml borosilicate bottles. Salinity was measured at the time of sampling using a refractometer. Bottles were stored at 3˚C and measured for AT within two weeks of sampling using an endpoint titration (Mettler Toledo® G20 Potentiometric Titrator). Methodological accuracy (within ±1%) of alkalinity titrations were verified and calibrated using Dr. Andrew Dickson’s (University of California San Diego, Scripps Institution of Oceanography) certified reference material for AT in seawater. The partial pressure of CO2 (pCO2,; μatm) was calculated in CO2SYS (V2.1, http://cdiac.ornl.gov/ftp/co2sys) based on measured AT, pHNBS, temperature, and salinity using K1 and K2 constants from Mehrbach et al. (1973)  refit by Dickson and Millero (1987)  and Dickson (1990) for KHSO4.

Field sampling and experimental designs:
Collections of wild, spawning ripe Atlantic silversides were made during high tide 1-3 days prior to full or new moons during the species spawning season. Adults were caught with a 30 m × 2 m beach seine from local salt marshes and transported live to our laboratory facilities. Ripe adults were held overnight at 20°C in well aerated tanks at low densities with no food and strip spawned the next day.

For each experiment, eggs from 20+ running-ripe females were gently mixed into shallow plastic dishes lined with 1 mm plastic window screening. 20+ males were stripped-spawned together into 500 ml glass beakers, mixed with seawater, stirred, then gently poured into spawning dishes and mixed with eggs for ~15 minutes. Screens were rinsed with seawater to remove unfertilized eggs and then soaked in a 100 ppm buffered iodine (Ovadine®) solution for 15 minutes to prevent fungal infection. Experiments were initiated within two hours of fertilization when replicate rearing vessels received precisely 100 embryos. Vessels were filled with clean seawater (filtered to 1 µm and UV sterilized). Optimal salinity (27-31) and light conditions (15 h light:9 h dark) for rearing M. menidia were maintained across experiments. Upon hatching larvae were immediately provided ad libitum rations of newly hatched brine shrimp nauplii (Artemia salina, San Francisco strain, brineshrimpdirect.com) and equal rations of powdered weaning diet (Otohime Marine Fish Diet, size A1, Reed Mariculture®). To quantify hatching survival, one day post first hatch larvae were counted by gently scooping small groups into replacement rearing vessels. For initial hatch measurements, random sub-samples (N = 10) from each replicate were preserved in 5% formaldehyde/freshwater solution buffered with saturated sodium tetraborate. All experiments were terminated when larvae reached ~10 mm standard length (SL). At termination, all survivors were counted and measured for standard length (SL, nearest 0.01 mm) via calibrated digital images (Image Pro Premier® V9.0).


Processing Description

BCO-DMO Processing:
- modified parameter names to conform with BCO-DMO naming conventions (replaced spaces with underscores);
- changed date format from mm/dd/yyyy to yyyy/mm/dd;
- replaced "n/a" with "nd";
- replaced spaces with underscores in columns: species, adult_collection_site;
- removed commas from adult_collection_site field;
- replaced original lat/lon values with decimal degree values provided by PI.


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Related Publications

Dickson, A. G. (1990). Standard potential of the reaction: , and and the standard acidity constant of the ion HSO4− in synthetic sea water from 273.15 to 318.15 K. The Journal of Chemical Thermodynamics, 22(2), 113–127. doi:10.1016/0021-9614(90)90074-z https://doi.org/10.1016/0021-9614(90)90074-Z
Methods
Dickson, A. G., & Millero, F. J. (1987). A comparison of the equilibrium constants for the dissociation of carbonic acid in seawater media. Deep Sea Research Part A. Oceanographic Research Papers, 34(10), 1733–1743. doi:10.1016/0198-0149(87)90021-5
Methods
Mehrbach, C., Culberson, C. H., Hawley, J. E., & Pytkowicx, R. M. (1973). MEASUREMENT OF THE APPARENT DISSOCIATION CONSTANTS OF CARBONIC ACID IN SEAWATER AT ATMOSPHERIC PRESSURE1. Limnology and Oceanography, 18(6), 897–907. doi:10.4319/lo.1973.18.6.0897
Methods
Murray, C., & Baumann, H. (2018). You Better Repeat It: Complex CO2 × Temperature Effects in Atlantic Silverside Offspring Revealed by Serial Experimentation. Diversity, 10(3), 69. doi:10.3390/d10030069
Results

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Parameters

ParameterDescriptionUnits
experimentExperiment number unitless
speciesScientific name of the speices (Menidia menidia, the Atlantic silverside) unitless
adult_collection_siteSite of the collection of wild adult spawners unitless
latitudeLatitude of adult collection site decimal degrees
longitudeLongitude of adult collection site decimal degrees
tankTank number unitless
replicateWithin experiment replicate number unitless
temperatureAverage temperature experienced by fish in degrees celsius degrees Celsius
pHAverage pH level experienced by the fish NBS pH NBS
pCO2Calculated average pCO2 levels in µatm calculated by CO2SYS based on alkalinity, pH, and temperature atm
fertilization_dateDate offspring were fertilized by stripspawning 20+ adults of each sex; formatted as yyyy/mm/dd unitless
hatch_sample_dateDate offspring were sampled for hatch survival and measurements; formatted as yyyy/mm/dd unitless
hatch_sample_ageAge in days post fertilization (dpf) on the date of hatch sampling days
final_sample_dateDate offspring were sampled for final survival and measurements; formatted as yyyy/mm/dd unitless
final_sample_ageAge in days post fertilization (dpf) on the date of final sampling days
rel_embryo_survivalrelative survival of embryos (0-1) unitless (fraction)
rel_larval_survivalrelative survival from hatch to experiment termination (0-1) unitless (fraction)
hatch_lengthAverage replicate hatch length (mm) millimeters (mm)
final_lengthAverage replicate final length (mm) millimeters (mm)
growth_rateaverage replicate growth rate (mm d-1) of offspring from hatch to experiment termination millimeters per day (mm d-1)


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Instruments

Dataset-specific Instrument Name
handheld pH probe
Generic Instrument Name
pH Sensor
Dataset-specific Description
Target pH levels were monitored daily using a handheld pH probes (Orion Ross Ultra pH/ATC Triode with Orion Star A121 pH Portable Meter; Intellical PHC281 pH Electrode with Hach HQ11D Handheld pH/ORP Meter) calibrated bi-weekly with 2-point pHNBS references.
Generic Instrument Description
General term for an instrument that measures the pH or how acidic or basic a solution is.

Dataset-specific Instrument Name
refractometer
Generic Instrument Name
Refractometer
Dataset-specific Description
Salinity was measured at the time of sampling using a refractometer.
Generic Instrument Description
A refractometer is a laboratory or field device for the measurement of an index of refraction (refractometry). The index of refraction is calculated from Snell's law and can be calculated from the composition of the material using the Gladstone-Dale relation. In optics the refractive index (or index of refraction) n of a substance (optical medium) is a dimensionless number that describes how light, or any other radiation, propagates through that medium.

Dataset-specific Instrument Name
Mettler Toledo G20 Potentiometric Titrator
Generic Instrument Name
Automatic titrator
Dataset-specific Description
Bottles were stored at 3˚C and measured for AT within two weeks of sampling using an endpoint titration (Mettler Toledo G20 Potentiometric Titrator). 
Generic Instrument Description
Instruments that incrementally add quantified aliquots of a reagent to a sample until the end-point of a chemical reaction is reached.

Dataset-specific Instrument Name
dissolved oxygen probe
Generic Instrument Name
Dissolved Oxygen Sensor
Generic Instrument Description
An electronic device that measures the proportion of oxygen (O2) in the gas or liquid being analyzed

Dataset-specific Instrument Name
airstone
Generic Instrument Name
Airstone
Dataset-specific Description
Elevated CO2 levels were achieved via gas proportioners (ColeParmer) mixing air with 100% CO2 (bone dry grade) that was delivered continuously to the bottom of each replicate rearing container via airstone.
Generic Instrument Description
Airstone - Also called an aquarium bubbler, is a piece of aquarium furniture, traditionally a piece of limewood or porous stone, whose purpose is to gradually diffuse air into the tank, eliminating the noise and large bubbles of conventional air filtration systems

Dataset-specific Instrument Name
glass submersible heaters
Generic Instrument Name
Immersion heater
Dataset-specific Description
Target treatment temperatures were controlled by thermostats (Aqualogic) which powered chillers (DeltaStar) or glass submersible heaters to maintain water bath temperatures.
Generic Instrument Description
Submersible heating element for water tanks and aquaria.

Dataset-specific Instrument Name
DeltaStar chiller
Generic Instrument Name
Aquarium chiller
Dataset-specific Description
Target treatment temperatures were controlled by thermostats (Aqualogic) which powered chillers (DeltaStar) or glass submersible heaters to maintain water bath temperatures.
Generic Instrument Description
Immersible or in-line liquid cooling device, usually with temperature control.

Dataset-specific Instrument Name
beach seine
Generic Instrument Name
Seine Net
Dataset-specific Description
Adults were caught with a 30 m × 2 m beach seine from local salt marshes and transported live to our laboratory facilities. 
Generic Instrument Description
A seine net is a very long net, with or without a bag in the centre, which is set either from the shore or from a boat for surrounding a certain area and is operated with two (long) ropes fixed to its ends (for hauling and herding the fish). Seine nets are operated both in inland and in marine waters. The surrounded and catching area depends on the length of the seine and of the hauling lines. (definition from: fao.org)


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Deployments

Mumford_Cove_Subsurface_Buoy

Website
Platform
Avery_Point
Start Date
2015-04-04
Description
Local subsurface buoy in Mumford Cove, CT, a shallow, coastal embayment in outer Long Island Sound, US Atlantic coast.

Poquot_Beach

Website
Platform
Avery_Point


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Project Information

Collaborative research: Understanding the effects of acidification and hypoxia within and across generations in a coastal marine fish (HYPOA)

Coverage: Eastern Long Island Sound, CT, USA


Description from NSF award abstract:
Coastal marine ecosystems provide a number of important services and resources for humans, and at the same time, coastal waters are subject to environmental stressors such as increases in ocean acidification and reductions in dissolved oxygen. The effects of these stressors on coastal marine organisms remain poorly understood because most research to date has examined the sensitivity of species to one factor, but not to more than one in combination. This project will determine how a model fish species, the Atlantic silverside, will respond to observed and predicted levels of dissolved carbon dioxide (CO2) and oxygen (O2). Shorter-term experiments will measure embryo and larval survival, growth, and metabolism, and determine whether parents experiencing stressful conditions produce more robust offspring. Longer-term experiments will study the consequences of ocean acidification over the entire life span by quantifying the effects of high-CO2 conditions on the ratio of males to females, lifetime growth, and reproductive investment. These studies will provide a more comprehensive view of how multiple stressors may impact populations of Atlantic silversides and potentially other important forage fish species. This collaborative project will support and train three graduate students at the University of Connecticut and the Stony Brook University (NY), two institutions that attract students from minority groups. It will also provide a variety of opportunities for undergraduates to participate in research and the public to learn about the study, through summer research projects, incorporation in the "Women in Science and Engineering" program, and interactive displays of environmental data from monitoring buoys. The two early-career investigators are committed to increasing ocean literacy and awareness of NSF-funded research through public talks and presentations.

This project responds to the recognized need for multi-stressor assessments of species sensitivities to anthropogenic environmental change. It will combine environmental monitoring with advanced experimental approaches to characterize early and whole life consequences of acidification and hypoxia in the Atlantic silverside (Menidia menidia), a valued model species and important forage fish along most of the US east coast. Experiments will employ a newly constructed, computer-controlled fish rearing system to allow independent and combined manipulation of seawater pCO2 and dissolved oxygen (DO) content and the application of static and fluctuating pCO2 and DO levels that were chosen to represent contemporary and potential future scenarios in productive coastal habitats. First CO2, DO, and CO2 × DO dependent reaction norms will be quantified for fitness-relevant early life history (ELH) traits including pre- and post-hatch survival, time to hatch, post-hatch growth, by rearing offspring collected from wild adults from fertilization to 20 days post hatch (dph) using a full factorial design of 3 CO2 × 3 DO levels. Second, the effects of tidal and diel CO2 × DO fluctuations of different amplitudes on silverside ELH traits will be quantified. To address knowledge gaps regarding the CO2-sensitivity in this species, laboratory manipulations of adult spawner environments and reciprocal offspring exposure experiments will elucidate the role of transgenerational plasticity as a potential short-term mechanism to cope with changing environments. To better understand the mechanisms of fish early life CO2-sensitivity, the effects of temperature × CO2 on pre- and post-hatch metabolism will be robustly quantified. The final objective is to rear silversides from fertilization to maturity under different CO2 levels and assess potential CO2-effects on sex ratio and whole life growth and fecundity.

Related references:
Gobler, C.J. and Baumann, H. (2016) Hypoxia and acidification in ocean ecosystems: Coupled dynamics and effects on marine life. Biology Letters 12:20150976. doi:10.1098/rsbl.2015.0976

Baumann, H. (2016) Combined effects of ocean acidification, warming, and hypoxia on marine organisms. Limnology and Oceanography e-Lectures 6:1-43. doi:10.1002/loe2.10002

Depasquale, E., Baumann, H., and Gobler, C.J. (2015) Variation in early life stage vulnerability among Northwest Atlantic estuarine forage fish to ocean acidification and low oxygen Marine Ecology Progress Series 523: 145–156.doi:10.3354/meps11142



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Funding

Funding SourceAward
NSF Division of Ocean Sciences (NSF OCE)

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