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MEPS just published our most recent paper on sand lance CO2-sensitivity!

A potential ripple effect from carbon in the atmosphere could have severe impacts throughout the ocean ecosystem

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This photo shows sand lance embryos that have and have not hatched. Sand lance have trouble hatching at future ocean CO2 levels (photo courtesy of Emma Cross).


By Elaina Hancock. Reposted from UConn Today, 7 April 2022

When carbon is emitted into the atmosphere, about a quarter of it is absorbed by the earth’s oceans. As the oceans serve as a massive ‘sink’ for carbon, there are changes to the water’s pH – a measure of how acidic or basic water is. As oceans absorb carbon, their water becomes more acidic, a process called ocean acidification (OA). For years, researchers have worked to understand what effect this could have on marine life.

While most research so far shows that fish are fairly resilient to OA, new research from UConn, the University of Washington, the National Oceanic and Atmospheric Administration (NOAA), and Southern Connecticut State University, shows that an important forage fish for the Northwest Atlantic called sand lance is very sensitive to OA, and that this could have considerable ecosystem impacts by 2100. The team’s findings have just been published in Marine Ecology Progress Series 687.

Sand lance spawn in the winter months in offshore environments that tend to have stable, low levels of CO2, explains UConn Department of Marine Sciences researcher and lead author Hannes Baumann.

“Marine organisms are not living in a uniform ocean,” Baumann says. “In near shore environments, large CO2 fluctuations between day and night and between seasons are the norm, and the fish and other organisms are adapted to this variability. When we stumbled upon sand lances we suspected they are different. We thought that a fish that lives in a more open-ocean offshore environment might be more sensitive than the near-shore fish because there’s just much less variability.”

The project was a collaboration with physical oceanographers, including Assistant Professor of Marine Sciences Samantha Siedlecki and Michael Alexander from NOAA’s Physical Sciences Laboratory in Boulder, Colorado, who modeled CO2 levels in 2050 and 2100 for a specific part of the Gulf of Maine where sand lance spawn. Then Baumann and his team reared sand lance embryos in the lab under experimentally higher CO2 levels matching the projected levels.

There are instances of direct fish mortality as result of elevated CO2, but they are rare, says Baumann. However, sand lance embryos proved to be exceptionally sensitive, and fewer embryos hatched under future oceanic CO2 conditions. The researchers repeated the experiments three more times to avoid jumping to conclusions but each time they observed the same result.

“We found that embryo survival-to-hatch decreased sharply with increasing CO2 levels in the water, concluding that this is one of the most CO2-sensitive fish species studied thus far,” Baumann says.

Sand lances are surely one of the most important forage fish here on the Northwest Atlantic shelf… The humpback whales, sharks, tuna, cod, shearwaters, terns — you name it — they are all relying on sand lance.

With this interdisciplinary approach combining model forecasts and serial experimentation the researchers arrived at a picture that is much more specific.

“We consequently applied principles of serial experimentation, which is a most timely and important topic in ocean acidification research right now,” Baumann says. “Because our findings are backed up by repeated independent evidence, they are more robust than many published ocean acidification studies to date.”

In addition to preventing many sand lance embryos from developing normally, the researchers document a second negative, and novel, response to elevated CO2. Higher CO2 levels appear to make it harder for embryos to hatch.

Baumann explains the lowered pH likely renders enzymes needed for successful hatching less effective, leaving the embryos unable to break through their eggshell (chorion) to hatch.

The results show that by 2100, due to acidification, sand lance hatching success could be reduced to 71% of today’s levels. Since sand lance are such a critical component of the food web of the Northwest Atlantic, this marked decrease in sand lance would have profound impacts throughout the ecosystem.

“Sand lances are surely one of the most important forage fish here on the Northwest Atlantic shelf,” Baumann says. “Their range spans from the Mid Atlantic Bight all the way to Greenland. Where we studied them, on Stellwagen Bank, they are called the backbone of the ecosystem. The humpback whales, sharks, tuna, cod, shearwaters, terns — you name it — they are all relying on sand lance, and if sand lance productivity goes down, we will see ripple effects to all these higher trophic animals. Even though we humans don’t fish for sand lance, we need to take care of the species because it has such a huge effect on everything else.”

Baumann says this study supports the hypothesis that offshore, high latitude marine organisms like the sand lance may be among the most vulnerable to OA. As a result, these organisms and food webs will likely be impacted first and soon, and we must act now.

Previous research has focused on opportunistically chosen species when testing their sensitivity for ocean acidification, says Baumann, but this should change.

“We need strategic thinking about what species we are testing next, because we cannot test every marine fish species, that’s an impossible task. We should concentrate on fish species that are likely the most vulnerable, and therefore the ones that are probably being affected first and this research makes a compelling argument that those are the fish species at higher latitudes and in more offshore than nearshore environments.”


Kelli Mosca presents Master thesis research on Atlantic Sturgeon

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21 March 2022. Today, Baumann lab graduate student Kelli Mosca presented her Masters thesis entitled "Atlantic sturgeon (Acipenser oxyrinchus) Growth and Habitat Use in the Connecticut River and Long Island Sound". She did a marvelous job summarizing the multifaceted findings on age and growth of Atlantic sturgeon in the eastern Long Island Sound and the Connecticut River, their movement patterns based on analyses of acoustic telemetry data, while evaluating the evidence for indications that Atlantic sturgeon may utilize the Connecticut River again for spawning.

Kelli was an inspirational and cherished member of the Baumann lab, who literally mastered the challenges of being a 'whole-pandemic' Master student. Her dedication and continued work for CTDEEP were awarded by an offer for a CTDEEP Fishery Biologist I position, which she has wholeheartedly accepted. Congratulations Kelli, and all the best for the next steps in your career!

The UConn Department of Marine Sciences
Presents a Master’s Thesis Presentation by

Kelli Mosca
B.S., University of New Haven, 2017

12:00 p.m., Monday, March 21, 2022
Lowell Weicker Building, Seminar Room 103 or Via WebEx

Atlantic sturgeon (Acipenser oxyrinchus) Growth and Habitat Use in the Connecticut River and Long Island Sound

Atlantic sturgeon (ATS, Acipenser oxyrinchus) are long-lived, anadromous, and endangered fish with a wide geographical distribution along the east coast of North America. Historically known to spawn in numerous rivers, many spawning runs ended due to intense fishing pressure and habitat obstruction in the 19th and 20th centuries. This was thought to be true for the longest river in the US Northeast, the Connecticut River, until pre-migratory ATS juveniles appeared in the river in 2014. Here, I use a long-term archive of fin spine samples and three years of acoustically tagged ATS to generally expand knowledge about the ATS using the CT River and LIS, and specifically examine these empirical data for potential evidence of re-emergent spawning behavior. I analyzed 301 sections of ATS fins spines collected from 1988-2021 to determine age, annuli widths, and thus population- and individual-based growth patterns. I found that the vast majority of ATS in my study area were juveniles and sub-adults with an average (± SD) age of 7.5 ± 3.1 years and an average (± SD) length of 101 ± 26 cm. The weighed, population-based Von Bertalanffy growth model estimated a K of 0.08 (95% CL, 0.01/0.17) and a L∞ of 171.2 cm (95% CI, 129/547 cm), the latter likely showing signs of missing large adults. K and L∞ distributions showed no sign of sex-specific multi-modality. Longitudinal length back-calculations revealed the selective disappearance of faster growing phenotypes (at ages 2-6) with increasing age at capture, which is clear evidence for Lee’s phenomenon. Acoustic detections of telemetered Atlantic sturgeon (2019-2021) revealed that most sturgeon in 2019 and 2020 utilized the Lower CT River (brackish water), whereas in 2021 detections were highest in LIS (salt water). Detections in the Upper CT River (freshwater) were common but much less dense across years, with 53%, 69% and 45% of ATS detected in the Upper CT River at some point in each season (2019-2021 respectively). I found a positive relationship of fish proportion in the CT River with temperature, but an inverse relationship of fish proportion in the CT River with river discharge. On average, the arrival of fish in the CT River occurred in June, when water temperatures were 17.5 - 24.9 ºC, while the departure from the CT River generally occurred in October, coinciding with river temperatures of 15.2 - 20.4 ºC. Some of the fish utilizing the Upper CT River made directed movements to a potential spawning ground at Portland, CT (river km 47). However, these movements occurred in mid- to late August (12th -23rd), which is inconsistent with the typical spring timing of ATS spawning runs in northern populations. Fall spawning runs are only known for southern ATS populations. In addition to timing, ATS sizes in the Upper CT River also do not support spawning behavior, because fish of all sizes (72 – 154 cm TL) and ages (3-15) visited the Portland area for 0.25 – 63.25 days. I conclude that neither age nor telemetry data support the re-emergence of the CT River as an ATS spawning ground. Future work will benefit from a more even sampling of gear sizes and should examine possible explanations for ATS freshwater utilization including feeding and individual preferences.

Major Advisor: Hannes Baumann
Associate Advisor: Eric Schultz
Associate Advisor: Tom Savoy
Associate Advisor: Jacque Benway
Associate Advisor: Catherine Matassa

Lucas Jones presents his Masters Thesis research!

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Lucas and Hannes celebrating the successful thesis presentation

Monday, November 22nd 2021. Big and heartfelt congratulations to Lucas Jones, who presented his Master thesis to his peers at the institute and colleagues national and international. Well done, Lucas!

A link to his recorded presentation will be posted here soon.


The UConn Department of Marine Sciences

Presents a Master’s Thesis Presentation by

Lucas Jones

B.A., University of Connecticut, 2018

4:00 p.m., Monday, November 22, 2021

Marine Sciences Building, Seminar Room 103

 

Using Low-Coverage, Whole Genome Sequencing to Study Northern Sand Lance (Ammodytes dubius) Population Connectivity in the Northwest Atlantic

 Northern sand lance (Ammodytes dubius) are key forage fish in Northwest Atlantic (NWA) shelf ecosystems, where they exclusively occur on coarse-grain, offshore sand banks. This patchy occurrence may result in genetically more fragmented, less connected populations, but traditional morphological or genomic approaches have so far been unsuccessful in fully resolving the species’ population structure and connectivity. My study pursued an alternative genomic approach, using low-coverage, whole genome sequencing (LcWGS) to address these important questions. I extracted DNA from 273 A.dubius specimens collected by collaborators from sevenregions across the species geographical range, from Greenland to New Jersey, USA. From LcWGS data, I identified 11,558,126 single nucleotide polymorphisms (SNPs) that allowed quantifying genetic differentiation between populations (FST), thereby revealing the genetic structuring of populations throughout the NWA. Despite the potentially homogenizing influence of the general north to south ocean circulation, I found a clear genetic break around Nova Scotia that delineated a northern from a southern A. dubius supergroup. Only within the southern supergroup, genetic distances increased with the geographic distance between sample sites. At the focal site of Stellwagen Bank (southern Gulf of Maine), A. dubius samples collected over several years (2014 – 2019) revealed small but significant temporal genetic differences that imply varying occupation of this offshore habitat by genetically different sand lance contingents. Inclusion of samples from the inshore congener A. americanus confirmed the clear genetic separation between both species and further determined that all sand lance caught on Stellwagen Bank are exclusively A. dubius. Overall, my work suggests the existence of two spatially distinct A. dubius populations with little ‘realized’ connectivity, which is critical knowledge to aid protection and management of offshore marine resources.

 

Major Advisor:                   Hannes Baumann

Associate Advisor:            Nina Overgaard Therkildsen

Associate Advisor:            Senjie Lin

ICES Journal of Marine Science publishes long-term fecundity study!

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2 November 2021. We are happy report that the ICES Journal of Marine Science just published the last major experimental paper on Atlantic silverside CO2-sensitivity from our lab. Callie Concannon and co-authors report on two complementary, long-term rearing trials in 2015/16 and 2018/19, where silverside juveniles or newly fertilized embryos were reared under contrasting temperature and CO2 conditions to maturity. This revealed negative effects of high CO2 conditions on female fecundity, but only at the warm, not the cold temperature treatments (Fig. below). Our study and its data are novel, because they were generated by the first whole-life CO2 rearing experiment of a fish and are the first empirical fecundity effects shown for a broadcast-spawning fish species.

The paper is also special to us, because its publication marks the erstwhile conclusion of our yearlong, NSF-funded efforts (OCE#1536165) to understand the CO2 sensitivity and its mechanisms in this important forage fish and long-standing model in fish ecology and evolution. The project ran from 2015 - 2020, produced 15 publications, 2 book chapters, and over 40 presentations, while furthering the careers of a post-doc, a PhD student, 5 Master students and over 10 undergraduates.


A day on the water with CTDEEPs sturgeon researchers!

28 September 2021. The day started nice enough but soon turned into dark threatening clouds, which the sturgeon researchers that day eyed with concern. Tom Savoy, Deb Pacileo, and Jacque Benway from CTDEEP, along with Kelli Mosca, Steve, Jake, and Hannes started to cast off late morning and slowly motored up the Connecticut River. "We're trying to catch the slack tide to set our sturgeon gill nets" explained Tom, the veteran sturgeon researcher, who has accompanied and steered most monitoring and protection efforts of these iconic fish over the past decades. It was Tom and his colleagues, who in 2014 caught the first baby Atlantic sturgeon in Connecticut River - a potential sign for a long hoped for recovery and the starting point of our project funded by CT SeaGrant.

The day trip was almost over, before it began. The downpour short but relentless and Jacque eyeing the lightening coming from the west with unease. But just before we could decide to fully head back, the radio call from the other boat that several sturgeon had indeed been caught in the gill nets! We therefore proceeded going through the routine measurement protocol that has been implemented for many years and is part of a federal permit to study this endangered species (No. 19641). Even a photograph is considered a sample, which is why we were glad to have our photographer Jacob Snyder (RedSkiesPhotography.com) fully accredited for the trip today. We saw with our own eyes, how a caught sturgeon was first being carefully pried loose from the gill-netting and placed in an aerated observation sump to prevent stress. All individuals are then checked for previously inserted tags using a specialized scanner (1) and if none is found, a new PIT-tag is inserted under the skin with the help of a syringe (3). All sturgeon are also measured for length (2), gape (5) and head width, and weighed (6) before being released back into the water. Tissue samples and in a limited number of fish also fin spine samples are taken for genetic information and growth data, the latter Kelli analyzed for her Masters Thesis.

The potential re-emergence of Atlantic sturgeon spawning in the Connecticut River is a success story of research and conservation, however, these efforts need to be sustained and widely communicated for the success to endure.

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1. On 28 September 2021, a juvenile Atlantic sturgeon caught in the Connecticut River is being checked for any previously implemented tags 2. Juvenile or sub-adult sturgeon are being measured for fork and total length using a measuring board 3. CTDEEP researcher Jacque Benway inserts a PIT tag into an sub-adult Atlantic sturgeon 4. Subadult sturgeon on a wet tarp before being released into wild again 5. All sturgeon are measured for mouth gape and head width 6. All sturgeon are being weighed with a gentle contraption 7. Master student Kelli Mosca with an adult sturgeon (all images by Jacob Snyder, RedSkiesPhotography)

44th Larval Fish Conference held virtually 24-26 June

Groton, CT 24-26 June 2021. The long awaited and anxiously prepared virtual 44th Larval Fish Conference was held, featuring more than 240 participants from 28 countries. 58 scientific talks, including 3 keynote lectures were given via Cisco’s WebEx platform, whereas networking activities such as poster presentations, ‘Meet the Speaker’ events, and Mentor hours used the innovative Gatherly platform. The technology was working out well, the preparation paid off, and delegates were overall enthusiastic about this virtual alternative, which was forced on us by Covid-19, but may have shown us new ways and concepts to broaden the societies reach and equality.

The post-conference website is housed at https://lfc44.marinesciences.uconn.edu

Special thanks go to the scientific steering committee Eric Schultz, Jacqueline Webb, and Paul Anderson. Lauren Schaller, Anne Hill, Harley Erickson, and Kate Copeland from UConn’s conference services did a great job as well preparing and running parts of the events. Support came from NOAA’s Northeast Fisheries Science Center.


Our sandlance work featured in the CapeCodFishermen

Reposted from TheCapeCodFishermen, April 28th 2021

By David N. Wiley

Bluefin tuna and striped bass crash through the waves. Seabirds wheel overhead and plunge into the water. Gape-mouthed whales rise from below. Schools of cod and dogfish hide below the surface.

While the convergence of such diverse sea life might seem accidental, those in the know thank a small, slender fish called a sand eel for the bonanza.

Also known as sand lance, these three-to-six inch forage fish are a main food source for many of the top predators in the Gulf of Maine and on Georges Bank, including some of the most commercially important species.

As their name implies, sand lance are tied to sand habitat, but not just any sand will do. To avoid predators, sand lance spend most of the night and parts of the day buried. When disturbed, they rocket out of the bottom, then dive head first and at full speed back into the sand.

As a result, their sand of choice has to be coarse enough to hold oxygen for the fish to “breathe” while buried, but soft enough to allow high-speed body penetration. One of the reasons Cape Cod is their Mecca is a band of perfect sand stretching from Stellwagen Bank along the backside of Cape Cod, past Chatham and up through Georges Bank. Whether you are a fisherman, whale watcher or seabird enthusiast, it’s this band of sand, and the sand lance that inhabit it, that makes the Cape special.

Sand and sand lance are the backbone of Stellwagen Bank National Marine Sanctuary, responsible for it being one of the top places in the United States for viewing marine life, and a centuries old, highly productive fishing ground. Yet while fishermen appreciate the importance of sand lance, little is known about their biology and most of the world does not know they exist.

To remedy the situation, a team of researchers led by scientists from Stellwagen Bank National Marine Sanctuary with partners from Boston University, Center for Coastal Studies, University of Connecticut, U.S. Geological Survey and Woods Hole Oceanographic Institution have been studying the forage fish to determine its importance and unlock some of its secrets.

One of the project’s first goals was to identify the sand lance spawning season. Using a specially designed and permitted small-mesh trawl, fished from Steve Welch’s F/V Mystic or NOAA’s R/V Auk, the team captured and examined sand lance. Thought to spawn from late fall through winter, several years of work demonstrated that sand lance on Stellwagen Bank spawn in a very narrow window at the end of November. Eggs are deposited on the seafloor and hatch after approximately six weeks.

Then things get interesting. Once hatched, sand lance are tiny, free-floating larvae for two to three months. Given this long free-floating period and the currents flowing over Stellwagen Bank, many sand lance born on the bank cannot stay there. So where do they come from and where do their offspring go?

To answer this question, the team used hydrographic modeling to backtrack to where free floating particles (like larval sand lance) would have originated prior to their sand settlement in March or April, and where drifting particles would end up two or three months after hatching.

It appears that larval sand lance settling on Stellwagen originate off the coast of Maine; years of highest sand lance abundance correspond to conditions that would have transported additional larval sand lance from as far north as Nova Scotia. The same modeling indicated that larval sand lance originating on Stellwagen Bank transport south to the Great South Channel and Nantucket Shoals (but not Georges Bank). In some years, currents moved them as far as New Jersey.

This is just another example of the interconnected world that creates a productive marine environment. Since few sand lance in the study lived past three years, the dependence on shifting currents to populate the bank could be one thing responsible for boom and bust years typical of sand lance abundance. The team is currently examining genetics of sand lance taken from throughout the Gulf of Maine, the mid-Atlantic, and eastern Canada, to gain additional insight into population structure.

Do boom-bust years influence the distribution and abundance of predators? The team investigated the association of sand lance with humpback whales and great shearwater seabirds by placing satellite tags on both species to track their movements.

Throughout the Gulf of Maine, tracking revealed that both species spend the vast majority of their time over sand lance habitat, and DNA from fecal shearwater samples showed sand lance to be the bird’s main prey. Surveys in Stellwagen also demonstrated a high co-occurrence of sand lance, humpback whales and great shearwaters.

Sand lance feed primarily from February to July, mostly on Calanus finmarchicus copepods. They stop feeding from August through October, with low levels of feeding from the end of November to January. Body growth and fat content show similar trends, with length and fat stores increasing from February to July. After July, the fish retreat to bury in the sandy bottom, conserving energy for spawning.

The team then turned its attention to the future of the valuable fish, something of extreme importance to fishermen. Ripe fish captured in November were strip-spawned on board the boats and transported to Connecticut, where eggs and larvae were raised in special tanks that allowed temperature and acidity to be manipulated to mimic future ocean conditions under climate change. Increased temperature and acidity had a dramatic negative impact on larval survival. According to Dr. Hannes Baumann, whose lab led the work, sand lance may be unusually sensitive to ocean acidification.

The future of sand lance was also a focus of team members Joel Llopiz and Justin Suca from Woods Hole Oceanographic Institution. They came to some worrisome conclusions.

The abundance of tiny C. finmarchicus copepods directly influences sand lance health: Abundant C. finmarchicus led to good parental condition and high reproductive success, while low numbers resulted in poor parental condition and poor reproductive success. Scientists have suggested climate change scenarios in the Gulf of Maine will lead to reduced abundance of this critical copepod resource. Adding to the problem was their finding that warm slope water coming through the Northeast Channel north of Georges Bank led to the death of overwintering reproductive adults.

With the Gulf of Maine warming faster than 99 percent of the world’s oceans, there is concern about the future of sand lance and its potential impact to the productivity of the Gulf of Maine, Georges Bank and other areas. While states with fisheries and other marine resources supported by sand lance cannot solve climate change issues, they can work to make sand lance more resilient to climate change. One way is to eliminate as many non-climate stressors as possible.

For example, in 2020 Massachusetts promulgated a rule limiting daily sand lance landings to 200 pounds. Rhode Island followed suit in 2021. These rules were designed to discourage the development of a commercial fishery for the species, such as the huge industrial fishery in Europe’s North Sea.

Since a commercial sand lance fishery does not currently exist here, adopting this rule by other states would be an easy, proactive way to make our waters, and the people who depend on them, more resistant to climate change disruption.

(Dr. David N. Wiley is the Research Ecologist for Stellwagen Bank National Marine Santuary. Funding for the project was provided by the Bureau of Ocean Energy Management, The Volgenau Foundation, Northeast and Woods Hole Sea Grant, International Fund for Animal Welfare, Stellwagen Bank National Marine Sanctuary and the National Marine Sanctuary Foundation. Dan Blackwood, Dr. Gavin Fay, Peter Hong, Dr. Les Kaufmann, Kevin Powers, Dr. Jooke Robbins, Dr. Tammy Silva, Mike Thompson, and Dr. Page Valentine contributed to the study)

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‘Hooked on OA’ – Hannes talks about fish CO2 sensitivity to recreational anglers

25 February 2021. The Mid-Atlantic Ocean Acidification Network (MACAN) organized a four-part webinar series on Ocean Acidification geared specifically towards recreational anglers and shellfish collectors in the Mid-Atlantic region. The series is called “Hooked on OA” and invited Hannes on this February Thursday to explain the state of OA science particularly for fishes. A big thanks to the organizers and the more than 50 people who participated in this webinar.

HookedonOA


If you missed it and are still interested, you can watch the Zoom webinar here:

[Lab news] Adult black sea bass arrive at UConn’s Rankin lab

Establishing a Black Sea Bass brood stock to study early life history effects of warming and acidification

Max-Zawell
Max Zavell

December 2020. We are happy to announce that as of a few weeks, we are now housing two brood stock populations of Black Sea Bass (Centropristis striata) at UConn's Department of Marine Sciences Rankin Seawater Lab. The adults will be reared for the next few months under contrasting CO2 regimes in 1000gal tanks each. After photoperiod manipulation and hormone-injection we plan to strip-spawn adult fish and test their offspring for their CO2 and temperature sensitivity. Black Sea Bass are rapidly increasing in abundance in New England Water and the Gulf of Maine, but research on early life stages is still lacking. PhD student Max Zavell meticulously taking care of these fish now.

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Catch a glimpse of these beautiful animals below

JEB publishes paper on metabolic effects of high CO2 in silverside embryos! [New publication]

19 November 2020. We are happy to announce that the Journal of Experimental Biology just published the latest paper on CO2 effects in the early life stages of Atlantic silversides! For her PhD research at Stony Brook University, Teresa meticulously measured oxygen consumption in developing silverside embryos and newly hatched larvae exposed to contrasting oxygen and CO2 conditions throughout multiple experiments in 2017 and 2018. Her work shows that the metabolism of embryos but not larvae is sensitive to elevated CO2 conditions, leading to higher metabolic rates at normoxic levels, but reduced metabolic rates under low oxygen conditions, compared to controls. These basic empirical data confirm the emerging picture that CO2 effects in marine fish manifest largely if at all during early ontogeny, i.e., during the embryo stages. Well done, Teresa, and congratulations to your first lead-author paper!
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Conceptual diagram of the relationship between PO2 and RMR of M. menidia embryos in ambient and elevated PCO2. Hypothesized shifts in the relationship between embryonic RMR and PO2 are shown for elevated (orange) versus ambient (blue) PCO2. Our results (measured at the PO2 levels marked by black dots) suggest that PCO2 can influence both the critical oxygen partial pressure (Pcrit, gray lines) and the oxygen-independent RMR. At higher PO2 levels, RMR increases with PCO2, potentially owing to increased metabolic demand. As PO2 decreases, embryonic RMR reaches Pcrit and becomes oxygen dependent at a higher PO2 level in acidified than in ambient PCO2 conditions. Low intracellular red blood cell pH caused by high PCO2 can be expected to reduce hemoglobin–O2 affinity (Bohr effect) and make embryonic RMR less hypoxia resistant, which could manifest as an increase in Pcrit for embryos in elevated PCO2.