Marine Biology

, 166:62 | Cite as

Dose-dependence and small-scale variability in responses to ocean acidification during squid, Doryteuthis pealeii, development

  • Casey ZakroffEmail author
  • T. Aran Mooney
  • Michael L. Berumen
Original paper


Coastal squids lay their eggs on the benthos, leaving them to develop in a dynamic system that is undergoing rapid acidification due to human influence. Prior studies have broadly investigated the impacts of ocean acidification on embryonic squid, but have not addressed the thresholds at which these responses occur or their potential variability. We raised squid, Doryteuthis pealeii (captured in Vineyard Sound, Massachusetts, USA: 41°23.370'N 70°46.418′W), eggs in three trials across the breeding season (May–September, 2013) in a total of six chronic pCO2 exposures (400, 550, 850, 1300, 1900, and 2200 ppm). Hatchlings were counted and subsampled for mantle length, yolk volume, hatching time, hatching success, and statolith morphology. New methods for analysis of statolith shape, rugosity, and surface degradation were developed and are presented (with code). Responses to acidification (e.g., reduced mantle lengths, delayed hatching, and smaller, more degraded statoliths) were evident at ~ 1300 ppm CO2. However, patterns of physiological response and energy management, based on comparisons of yolk consumption and growth, varied among trials. Interactions between pCO2 and hatching day indicated a potential influence of exposure time on responses, while interactions with culture vessel highlighted the substantive natural variability within a clutch of eggs. While this study is consistent with, and expands upon, previous findings of sensitivity of the early life stages to acidification, it also highlights the plasticity and potential for resilience in this population of squid.





Dorsal mantle length


Environmental Systems Laboratory


Kruskal–Wallis test


Linear regression


Marine Biological Laboratory


Ocean acidification


Scanning electron microscopy


Yolk volume



We would like to thank D. Remsen, the MBL Marine Resources Center staff, and MBL Gemma crew for their help acquiring squid. R. Galat and WHOI facilities staff provided system support. D. McCorkle, KYK Chan, and M. White provided guidance and insight into the acidification system and water quality monitoring. A. Solow provided statistics advice. We thank L. Kerr and the MBL Central Microscopy Facility for their aid with the SEM. We greatly appreciate E. Bonk, S. Zacarias, M. Lee, and A. Schlunk for their outstanding advice and assistance with this experiment. Thanks also to editors and anonymous reviewers for their constructive feedback on this manuscript.


This material was based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. 1122374 to CZ. This project was funded by National Science Foundation Grant No. 1220034 to TAM.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Supplementary material

227_2019_3510_MOESM1_ESM.pdf (5.5 mb)
Supplementary material 1 (PDF 5625 kb)


  1. Anthony KRN, Kline DI, Diaz-Pulido G, Dove S, Hoegh-Guldberg O (2008) Ocean acidification causes bleaching and productivity loss in coral reef builders. Proc Natl Acad Sci 105:17442–17446. CrossRefPubMedGoogle Scholar
  2. Arnold JM, Summers WC, Gilbert DL, Manalis RS, Daw NW, Lasek RJ (1974) A guide to laboratory use of the squid Loligo pealei. Marine Biological Laboratory, Woods HoleCrossRefGoogle Scholar
  3. Bartol IK, Krueger PS, Thompson JT, Stewart WJ (2008) Swimming dynamics and propulsive efficiency of squids throughout ontogeny. Integr Comp Biol 48:720–733. CrossRefPubMedGoogle Scholar
  4. Baumann H, Wallace RB, Tagliaferri T, Gobler CJ (2014) Large natural pH, CO2 and O2 fluctuations in a temperate tidal salt marsh on diel, seasonal, and interannual time scales. Estuar Coasts. CrossRefGoogle Scholar
  5. Birk MA, McLean EL, Seibel BA (2018) Ocean acidification does not limit squid metabolism via blood oxygen supply. J Exp Biol. CrossRefPubMedGoogle Scholar
  6. Bonhomme V, Picq S, Gaucherel C, Claude J (2013) Momocs: outline analysis using R. J Stat Softw 56:1–24. CrossRefGoogle Scholar
  7. Boyle PR, Pierce GJ, Hastie LC (1995) Flexible reproductive strategies in the squid Loligo forbesi. Mar Biol 121:501–508CrossRefGoogle Scholar
  8. Buresch KC, Maxwell MR, Cox MR, Hanlon RT (2009) Temporal dynamics of mating and paternity in the squid Loligo pealeii. Mar Ecol Prog Ser 387:197–203. CrossRefGoogle Scholar
  9. Byrne M (2011) Impact of ocean warming and ocean acidification on marine invertebrate life history stages: vulnerabilities and potential for persistence in a changing ocean. Ocean Mar Biol Annu Rev 49:1–42. CrossRefGoogle Scholar
  10. Caldeira K, Wickett ME (2003) Oceanography: anthropogenic carbon and ocean pH. Nature 425:365. CrossRefGoogle Scholar
  11. Clayton TD, Byrne RH (1993) Spectrophotometric seawater pH measurements: total hydrogen results. Deep Res 40:2115–2129CrossRefGoogle Scholar
  12. Cohen AL, Holcomb M (2009) Why corals care about ocean acidification: uncovering the mechanism. Oceanography 22:118–127. CrossRefGoogle Scholar
  13. Collins MA, Burnell GM, Rodhouse PG (1995) Reproductive strategies of male and female Loligo forbesi (Cephalopoda: Loliginidae). J Mar Biol Assoc UK 75:621–634CrossRefGoogle Scholar
  14. Colmers WF, Hixon RF, Hanlon RT, Forsythe JW, Ackerson MV, Wiederhold ML, Hulet WH (1984) Spinner cephalopods: defects of statocyst suprastructures in an invertebrate analogue of the vestibular apparatus. Cell Tissue Res. CrossRefPubMedGoogle Scholar
  15. Dickson AG (1990) Standard potential of the reaction: AgCl(s) + (1/2)H2(g) = Ag(s) + HCl(aq), and and the standard acidity constant of the ion HSO4 in synthetic sea water from 273.15 to 318.15 K. J Chem Thermodyn 22:113–127. CrossRefGoogle Scholar
  16. Dickson AG, Sabine CL, Christian JR (2007) Guide to best practices for ocean CO2 measurements. PICES Spec Publ 3:p191. CrossRefGoogle Scholar
  17. Doney SC, Fabry VJ, Feely RA, Kleypas JA (2009) Ocean acidification: the other CO2 problem. Ann Rev Mar Sci 1:169–192. CrossRefPubMedGoogle Scholar
  18. Fabry VJ, Seibel BA, Feely RA, Orr JC (2008) Impacts of ocean acidification on marine fauna and ecosystem processes. ICES J Mar Sci 65:414. CrossRefGoogle Scholar
  19. Gallager SM, Mann R, Sasaki GC (1986) Lipid as an index of growth and viability in three species of bivalve larvae. Aquaculture 56:81–103. CrossRefGoogle Scholar
  20. Gazeau F, Parker LM, Comeau S, Gattuso J-PP, O’Connor WA, Martin S, Pörtner H-O, Ross PM (2013) Impacts of ocean acidification on marine shelled molluscs. Mar Biol 160:2207–2245. CrossRefGoogle Scholar
  21. Gledhill DK, White MM, Salisbury J, Thomas H, Misna I, Liebman M, Mook B, Grear J, Candelmo AC, Chambers RC, Gobler CJ, Hunt CW, King AL, Price NN, Signorini SR, Stancioff E, Stymiest C, Wahle RA, Waller JD, Rebuck ND, Wang ZA, Capson TL, Morrison JR, Cooley SR, Doney SC (2015) Ocean and coastal acidification off New England and Nova Scotia. Oceanography 28:182–197. CrossRefGoogle Scholar
  22. Gray CL (1992) Long-finned Squid (Loligo pealei) Species Profile. Current Report: The Narragansett Bay Project NBP-92-106. Rhode Island Department of Environmental Management, Division of Fish & Wildlife, Marine Fisheries Section, pp 1–54Google Scholar
  23. Guerra Á, Allcock L, Pereira J (2010) Cephalopod life history, ecology and fisheries: an introduction. Fish Res 106:117–124. CrossRefGoogle Scholar
  24. Gutowska MA, Melzner F (2009) Abiotic conditions in cephalopod (Sepia officinalis) eggs: embryonic development at low pH and high pCO2. Mar Biol 156:515–519. CrossRefGoogle Scholar
  25. Haigh R, Ianson D, Holt CA, Neate HE, Edwards AM (2015) Effects of ocean acidification on temperate coastal marine ecosystems and fisheries in the northeast Pacific. PLoS One 10:e0117533. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Hanlon RT, Messenger JB (1998) Cephalopod behaviour. Cambridge University Press, CambridgeGoogle Scholar
  27. Hanlon R, Bidwell J, Tait R (1989) Strontium is required for statolith development and thus normal swimming behaviour of hatchling cephalopods. J Exp Biol 141:187–195PubMedGoogle Scholar
  28. Hofmann GE, Todgham AE (2010) Living in the now: physiological mechanisms to tolerate a rapidly changing environment. Annu Rev Physiol 72:127–145. CrossRefPubMedGoogle Scholar
  29. Hu MY, Sucre E, Charmantier-Daures M, Charmantier G, Lucassen M, Himmerkus N, Melzner F (2010) Localization of ion-regulatory epithelia in embryos and hatchlings of two cephalopods. Cell Tissue Res 339:571–583CrossRefGoogle Scholar
  30. Hu MY, Tseng Y-C, Stumpp M, Gutowska MA, Kiko R, Lucassen M, Melzner F (2011a) Elevated seawater pCO2 differentially affects branchial acid-base transporters over the course of development in the cephalopod Sepia officinalis. Am J Physiol Regul Integr Comp Physiol 300:R1100–R1114. CrossRefPubMedGoogle Scholar
  31. Hu MY, Tseng Y-C, Lin L-Y, Chen P-Y, Charmantier-Daures M, Hwang P-P, Melzner F (2011b) New insights into ion regulation of cephalopod molluscs: a role of epidermal ionocytes in acid-base regulation during embryogenesis. AJP Regul Integr Comp Physiol 301:R1700–R1709. CrossRefGoogle Scholar
  32. Hu MY, Lee J-R, Lin L-Y, Shih T-H, Stumpp M, Lee M-F, Hwang P-P, Tseng Y-C (2013) Development in a naturally acidified environment: Na+/H+-exchanger 3-based proton secretion leads to CO2 tolerance in cephalopod embryos. Front Zool 10:51. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Hu MY, Guh Y-J, Stumpp M, Lee J-R, Chen R-D, Sung P-H, Chen Y-C, Hwang P-P, Tseng Y-C (2014) Branchial NH4 +-dependent acid–base transport mechanisms and energy metabolism of squid (Sepioteuthis lessoniana) affected by seawater acidification. Front Zool 11:55. CrossRefGoogle Scholar
  34. Ikeda Y, Wada Y, Arai N, Sakamoto W (1999) Note on size variation of body and statoliths in the oval squid Sepioteuthis lessoniana hatchlings. J Mar Biol Assoc UK 79:757–759. CrossRefGoogle Scholar
  35. Jacobson LD (2005) Longfin inshore squid, Loligo pealeii, life history and habitat characteristics. NOAA Technical Memorandum NMFS-NE-193. U.S. Department of Commerce, National Oceanic and Atmospheric Administration, National Marine Fisheries Service. Northeast Fisheries Science Center, Woods Hole, pp 1–42Google Scholar
  36. Kaplan MB, Mooney TA, McCorkle DC, Cohen AL (2013) Adverse effects of ocean acidification on early development of squid (Doryteuthis pealeii). PLoS One 8:e63714. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Lacoue-Labarthe T, Réveillac E, Oberhänsli F, Teyssié JL, Jeffree R, Gattuso JP (2011) Effects of ocean acidification on trace element accumulation in the early-life stages of squid Loligo vulgaris. Aquat Toxicol 105:166–176. CrossRefPubMedGoogle Scholar
  38. Langsrud Ø (2003) ANOVA for unbalanced data: use type II instead of type III sums of squares. Stat Comput 13:163–167. CrossRefGoogle Scholar
  39. Laptikhovsky V, Nikolaeva S, Rogov M (2018) Cephalopod embryonic shells as a tool to reconstruct reproductive strategies in extinct taxa. Biol Rev 93:270–283. CrossRefPubMedGoogle Scholar
  40. Long MH, Mooney TA, Zakroff C (2016) Extreme low oxygen and decreased pH conditions naturally occur within developing squid egg capsules. Mar Ecol Prog Ser 550:111–119. CrossRefGoogle Scholar
  41. Martins RS, Roberts MJ, Chang N, Verley P, Moloney CL, Vidal EAG (2010a) Effect of yolk utilization on the specific gravity of chokka squid (Loligo reynaudii) paralarvae: implications for dispersal on the Agulhas Bank, South Africa. ICES J Mar Sci 67:1323–1335. CrossRefGoogle Scholar
  42. Martins RS, Roberts MJ, Vidal ÉAG, Moloney CL (2010b) Effects of temperature on yolk utilization by chokka squid (Loligo reynaudii d’Orbigny, 1839) paralarvae. J Exp Mar Biol Ecol 386:19–26. CrossRefGoogle Scholar
  43. Maxwell MR, Hanlon RT (2000) Female reproductive output in the squid Loligo pealeii: multiple egg clutches and implications for a spawning strategy. Mar Ecol Prog Ser 199:159–170. CrossRefGoogle Scholar
  44. McMahon JJ, Summers WC (1971) Temperature effects on the developmental rate of squid (Loligo pealei) embryos. Biol Bull 141:561–567CrossRefGoogle Scholar
  45. Mehrbach C, Culberson CH, Hawley JE, Pytkowicz RM (1973) Measurement of the apparent dissociation constants of carbonic acid in seawater at atmospheric pressure. Limnol Oceanogr 18:897–907. CrossRefGoogle Scholar
  46. Mesnil B (1977) Growth and Life Cycle of Squid, Loligo pealei and Illex illecebrosus, from the Northwest Atlantic. Selected Papers Number 2. International Commission for the Northwest Atlantic Fisheries, Dartmouth, Canada, pp 55–69Google Scholar
  47. Michaelidis B, Ouzounis C, Paleras A, Pörtner H-O (2005) Effects of long-term moderate hypercapnia on acid-base balance and growth rate in marine mussels Mytilus galloprovincialis. Mar Ecol Prog Ser 293:109–118. CrossRefGoogle Scholar
  48. Miller GM, Watson S-A, Donelson JM, McCormick MI, Munday PL (2012) Parental environment mediates impacts of increased carbon dioxide on a coral reef fish. Nat Clim Change 2:858–861. CrossRefGoogle Scholar
  49. Murray CS, Malvezzi A, Gobler CJ, Baumann H (2014) Offspring sensitivity to ocean acidification changes seasonally in a coastal marine fish. Mar Ecol Prog Ser 504:1–11. CrossRefGoogle Scholar
  50. Navarro MO, Bockmon EE, Frieder CA, Gonzalez JP, Levin LA (2014) Environmental pH, O2 and capsular effects on the geochemical composition of statoliths of embryonic squid Doryteuthis opalescens. Water. CrossRefGoogle Scholar
  51. Navarro MO, Kwan GT, Batalov O, Choi CY, Pierce NT, Levin LA (2016) Development of embryonic market squid, Doryteuthis opalescens, under chronic exposure to low environmental pH and [O2]. PLoS One 11:e0167461. CrossRefPubMedPubMedCentralGoogle Scholar
  52. Navarro MO, Parnell PE, Levin LA (2018) Essential market squid (Doryteuthis opalescens) Embryo Habitat: a baseline for anticipated ocean climate change. J Shellfish Res 37:601–614. CrossRefGoogle Scholar
  53. NOAA (2019) Squid, mackerel, and butterfish quota monitoring page. In: NOAA Fish.—Gt. Atl. Reg. Accessed 16 Mar 2019
  54. Oyarzun FX, Strathmann RR (2011) Plasticity of hatching and the duration of planktonic development in marine invertebrates. Integr Comp Biol 51:81–90. CrossRefPubMedGoogle Scholar
  55. Pecl GT, Jackson GD (2008) The potential impacts of climate change on inshore squid: biology, ecology and fisheries. Rev Fish Biol Fish 18:373–385. CrossRefGoogle Scholar
  56. Pecl GT, Moltschaniwskyj NA (2006) Life history of a short-lived squid (Sepioteuthis australis): resource allocation as a function of size, growth, maturation, and hatching season. ICES J Mar Sci 63:995–1004. CrossRefGoogle Scholar
  57. Pecl GT, Moltschaniwskyj NA, Tracey SR, Jordan AR (2004) Inter-annual plasticity of squid life history and population structure: ecological and management implications. Oecologia 139:515–524. CrossRefPubMedGoogle Scholar
  58. Pierrot D, Lewis E, Wallace DWR (2006) MS Excel program developed for CO2 system calculations. In: ORNL/CDIAC-105a. Carbon Dioxide Information Analysis Center, Oak Ridge National Laboratory, U.S. Department of Energy. Oak Ridge, Tennessee, pp 1–17.
  59. Pörtner H-O, Reipschlager A, Heisler N (1998) Acid-base regulation, metabolism and energetics in Sipunculus nudus as a function of ambient carbon dioxide level. J Exp Biol 201:43–55PubMedGoogle Scholar
  60. Radtke RL (1983) Chemical and structural characteristics of statoliths from the short-finned squid Illex illecebrosus. Mar Biol 76:47–54. CrossRefGoogle Scholar
  61. Robin JP, Roberts M, Zeidberg L, Bloor I, Rodriguez A, Briceño F, Downey N, Mascaró M, Navarro M, Guerra A, Hofmeister J, Barcellos DD, Lourenço SAP, Roper CFE, Moltschaniwskyj NA, Green CP, Mather J (2014) Transitions during cephalopod life history: The role of habitat, environment, functional morphology and behaviour. In: Vidal EAG (ed) Advances in cephalopod science: biology, ecology, cultivation and fisheries. Academic Press, Cambridge, pp 361–437CrossRefGoogle Scholar
  62. Rosa R, Seibel BA (2008) Synergistic effects of climate-related variables suggest future physiological impairment in a top oceanic predator. Proc Natl Acad Sci 105:20776–20780. CrossRefPubMedGoogle Scholar
  63. Rosa R, Trübenbach K, Repolho T, Pimentel M, Faleiro F, Boavida-Portugal J, Baptista M, Lopes VM, Dionísio G, Leal MC, Calado R, Pörtner HO (2013) Lower hypoxia thresholds of cuttlefish early life stages living in a warm acidified ocean. Proc R Soc B Biol Sci 280:20131695. CrossRefGoogle Scholar
  64. Rosa R, Trübenbach K, Pimentel MS, Boavida-Portugal J, Faleiro F, Baptista M, Dionísio G, Calado R, Pörtner HO, Repolho T (2014a) Differential impacts of ocean acidification and warming on winter and summer progeny of a coastal squid (Loligo vulgaris). J Exp Biol 217:518–525. CrossRefPubMedGoogle Scholar
  65. Rosa R, O’Dor R, Pierce G (2014b) Myopsid squids. Nova Science Publishers Inc, New YorkGoogle Scholar
  66. Schunter C, Welch MJ, Ryu T, Zhang H, Berumen ML, Nilsson GE, Munday PL, Ravasi T (2016) Molecular signatures of transgenerational response to ocean acidification in a species of reef fish. Nat Clim Change 6:1014–1018. CrossRefGoogle Scholar
  67. Schunter C, Welch MJ, Nilsson GE, Rummer JL, Munday PL, Ravasi T (2018) An interplay between plasticity and parental phenotype determines impacts of ocean acidification on a reef fish. Nat Ecol Evol 2:334–342. CrossRefPubMedGoogle Scholar
  68. Seibel BA (2015) Environmental physiology of the jumbo squid, Dosidicus gigas (d’Orbigny, 1835) (Cephalopoda: Ommastrephidae): Implications for changing climate. Am Malacol Bull 33:1–13CrossRefGoogle Scholar
  69. Seibel BA (2016) Cephalopod susceptibility to asphyxiation via ocean incalescence, deoxygenation, and acidification. Physiology 31:418–429. CrossRefPubMedGoogle Scholar
  70. Spady BL, Watson S, Chase TJ, Munday PL (2014) Projected near-future CO2 levels increase activity and alter defensive behaviours in the tropical squid Idiosepius pygmaeus. Biol Open 3:1063–1070. CrossRefPubMedPubMedCentralGoogle Scholar
  71. Steer MA, Pecl GT, Moltschaniwskyj NA (2003) Are bigger calamary Sepioteuthis australis hatchlings more likely to survive? A study based on statolith dimensions. Mar Ecol Prog Ser 261:175–182. CrossRefGoogle Scholar
  72. Steer M, Moltschaniwskyj N, Nichols D, Miller M (2004) The role of temperature and maternal ration in embryo survival: using the dumpling squid Euprymna tasmanica as a model. J Exp Mar Biol Ecol 307:73–89. CrossRefGoogle Scholar
  73. Vidal EAG, Haimovici M (1998) Feeding and the possible role of the proboscis and mucus cover in the ingestion of microorganism by rhynchoteuthion paralarvae (Cephalopoda: Ommastrephidae). Bull Mar Sci 63:305–316Google Scholar
  74. Vidal EAG, DiMarco FP, Wormuth JH, Lee PG (2002a) Influence of temperature and food availability on survival, growth and yolk utilization in hatchling squid. Bull Mar Sci 71:915–931Google Scholar
  75. Vidal EAG, DiMarco FP, Wormuth JH, Lee PG (2002b) Optimizing rearing conditions of hatchling loliginid squid. Mar Biol 140:117–127. CrossRefGoogle Scholar
  76. Villanueva R, Quintana D, Petroni G, Bozzano A (2011) Factors influencing the embryonic development and hatchling size of the oceanic squid Illex coindetii following in vitro fertilization. J Exp Mar Biol Ecol 407:54–62. CrossRefGoogle Scholar
  77. Wang ZA, Wanninkhof R, Cai W-J, Byrne RH, Hu X, Peng T-H, Huang W-J (2013) The marine inorganic carbon system along the Gulf of Mexico and Atlantic coasts of the United States: insights from a transregional coastal carbon study. Limnol Oceanogr 58:325–342. CrossRefGoogle Scholar
  78. White MM, McCorkle DC, Mullineaux LS, Cohen AL (2013) Early exposure of bay scallops (Argopecten irradians) to high CO2 causes a decrease in larval shell growth. PLoS One 8:2–9. CrossRefGoogle Scholar
  79. York CA, Bartol IK (2016) Anti-predator behavior of squid throughout ontogeny. J Exp Mar Biol Ecol 480:26–35. CrossRefGoogle Scholar
  80. Zakroff C, Mooney TA, Wirth C (2018) Ocean acidification responses in paralarval squid swimming behavior using a novel 3D tracking system. Hydrobiologia 808:83–106. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Massachusetts Institute of Technology-Woods Hole Oceanographic Institution Joint Program in Oceanography/Applied Ocean Science and EngineeringCambridgeUSA
  2. 2.Biology DepartmentWoods Hole Oceanographic InstitutionWoods HoleUSA
  3. 3.Division of Biological and Environmental Science and Engineering, Red Sea Research CenterKing Abdullah University of Science and TechnologyThuwalSaudi Arabia

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