Journal of Comparative Physiology B

, Volume 186, Issue 3, pp 297–311 | Cite as

Molecular, behavioral, and performance responses of juvenile largemouth bass acclimated to an elevated carbon dioxide environment

  • Clark E. DennisIII
  • Shivani Adhikari
  • Adam W. Wright
  • Cory D. SuskiEmail author
Original Paper


Aquatic hypercarbia, either naturally occurring or anthropogenically induced, can have extensive impacts on aquatic environments and resident organisms. While the impact of acute hypercarbia exposure on the behavior and physiology of fishes has been well studied, relatively little work has examined the physiological impact and acclimation capacity of fishes to chronic hypercarbia. To better understand the impacts of prolonged hypercarbia exposure, largemouth bass were held at ambient CO2 (13 mg L−1) and elevated CO2 (31 mg L−1; ≈21,000 µatm) for 58 days. Following this acclimation period, fish were subjected to three separate, yet complementary, experiments: (1) acute hypercarbia challenge of 120 mg L−1 CO2 for 1 h to quantify physiological and molecular responses; (2) hypercarbia avoidance challenge to compare CO2 agitation and avoidance responses; and (3) swim performance challenge to quantify burst swimming performance. Acclimation to 31 mg L−1 CO2 resulted in a significant constitutive upregulation of c-fos expression in erythrocytes, combined with significant constitutive expression of hsp70 in both gill and erythrocytes, relative to controls. Largemouth bass acclimated to elevated CO2 also had a reduced glucose response (relative to controls) following an acute CO2 exposure, indicating a reduced stress response to CO2 stressors. In addition, largemouth bass acclimated to elevated CO2 conditions required 50 % higher CO2 concentrations to illicit agitation behaviors and displayed prolonged burst swimming abilities in high CO2 environments relative to controls. Together, results demonstrate that largemouth bass exposed to chronic hypercarbia may possess a physiological advantage during periods of elevated CO2 relative to naïve fish, which may permit increased performance in hypercarbia.


Acclimation Behavior Hypercarbia Invasive species Performance Stress 



This work was supported by the Illinois Department of Natural Resources through funds provided by the USEPA’s Great Lakes Restoration Initiative (GLRI). Partial funding was also provided by the USFWS Federal Aid in Sport Fish Restoration Project F-69-R. We would like to thank Greg King and Jennifer Shen for field and laboratory assistance. Dr. Zachary A. Cheviron and Dr. Anthony C. Yannarell provided excellent comments on this manuscript. All work performed in this study conformed to the guidelines established by the Institutional Animal Care and Use Committee (IACUC) of the University of Illinois (Protocol # 13123).


  1. Angilletta MJ (2009) Thermal adaptation: a theoretical and empirical synthesis. Oxford University Press, OxfordCrossRefGoogle Scholar
  2. Anscombe FJ, Tukey JW (1963) The examination and analysis of residuals. Technometrics 5:141–160CrossRefGoogle Scholar
  3. Araki H, Schmid C (2010) Is hatchery stocking a help or harm?: evidence, limitations and future directions in ecological and genetic surveys. Aquaculture 308:S2–S11CrossRefGoogle Scholar
  4. Barton BA (2002) Stress in fishes: a diversity of responses with particular reference to changes in circulating corticosteroids. Integr Comp Biol 42:517–525CrossRefPubMedGoogle Scholar
  5. Bernier NJ, Randall DJ (1998) Carbon dioxide anaesthesia in rainbow trout: effects of hypercapnic level and stress on induction and recovery from anaesthetic treatment. J Fish Biol 52:621–637Google Scholar
  6. Blancheton JP (2000) Developments in recirculation systems for Mediterranean fish species. Aquacult Eng 22:17–31CrossRefGoogle Scholar
  7. Brauner C, Baker D (2009) Patterns of acid–base regulation during exposure to hypercarbia in fishes. In: Cardio-respiratory control in vertebrates. Springer, Berlin, p 43–63Google Scholar
  8. Brauner CJ, Seidelin M, Madsen SS, Jensen FB (2000) Effects of freshwater hyperoxia and hypercapnia and their influences on subsequent seawater transfer in Atlantic salmon (Salmo salar) smolts. Can J Fish Aquat Sci 57:2054–2064CrossRefGoogle Scholar
  9. Caldeira K, Wickett ME (2003) Oceanography: anthropogenic carbon and ocean pH. Nature 425:365CrossRefPubMedGoogle Scholar
  10. Chevin L, Lande R, Mace GM (2010) Adaptation, plasticity, and extinction in a changing environment: towards a predictive theory. PLoS Biol 8:e1000357CrossRefPubMedPubMedCentralGoogle Scholar
  11. Clingerman J, Bebak J, Mazik PM, Summerfelt ST (2007) Use of avoidance response by rainbow trout to carbon dioxide for fish self-transfer between tanks. Aquacult Eng 37:234–251CrossRefGoogle Scholar
  12. Cole JJ, Caraco NF, Kling GW, Kratz TK (1994) Carbon dioxide supersaturation in the surface waters of lakes. Science 265:1568–1570CrossRefPubMedGoogle Scholar
  13. Cole JJ, Prairie YT, Caraco NF, McDowell WH, Tranvik LJ, Striegl RG, Duarte CM, Kortelainen P, Downing JA, Middelburg JJ (2007) Plumbing the global carbon cycle: integrating inland waters into the terrestrial carbon budget. Ecosystems 10:172–185CrossRefGoogle Scholar
  14. Colt J, Orwicz K (1991) Modeling production capacity of aquatic culture systems under freshwater conditions. Aquacult Eng 10:1–29CrossRefGoogle Scholar
  15. Cooley SR, Doney SC (2009) Anticipating ocean acidification’s economic consequences for commercial fisheries. Environ Res Let 4:024007CrossRefGoogle Scholar
  16. Curran T, Franza BR Jr (1988) Fos and Jun: the AP-1 connection. Cell 55:395–397CrossRefPubMedGoogle Scholar
  17. Dahlberg ML, Shumway DL, Doudoroff P (1968) Influence of dissolved oxygen and carbon dioxide on swimming performance of largemouth bass and coho salmon. J Fish Res Board Can 25:49–70CrossRefGoogle Scholar
  18. Deigweiher K, Koschnick N, Portner HO, Lucassen M (2008) Acclimation of ion regulatory capacities in gills of marine fish under environmental hypercapnia. Am J Physiol Regul Integr Comp Physiol 295:R1660–R1670CrossRefPubMedGoogle Scholar
  19. Dennis CE III, Kates DF, Noatch MR, Suski CD (2014) Molecular responses of fishes to elevated carbon dioxide. Comp Biochem Physiol A. doi: 10.1016/j.cbpa.2014.05.013 Google Scholar
  20. Díaz F, Re AD, González RA, Sánchez LN, Leyva G, Valenzuela F (2007) Temperature preference and oxygen consumption of the largemouth bass Micropterus salmoides (Lacepede) acclimated to different temperatures. Aquacult Res 38:1387–1394CrossRefGoogle Scholar
  21. Dixson DL, Munday PL, Jones GP (2010) Ocean acidification disrupts the innate ability of fish to detect predator olfactory cues. Ecol Lett 13:68–75CrossRefPubMedGoogle Scholar
  22. Douglas CE, Michael FA (1991) On distribution-free multiple comparisons in the one-way analysis of variance. Commun Stat Theory Methods 20:127–139CrossRefGoogle Scholar
  23. Feely RA, Sabine CL, Hernandez-Ayon JM, Ianson D, Hales B (2008) Evidence for upwelling of corrosive “acidified” water onto the continental shelf. Science 320:1490–1492CrossRefPubMedGoogle Scholar
  24. Fivelstad S, Olsen AB, Kløften H, Ski H, Stefansson S (1999) Effects of carbon dioxide on Atlantic salmon (Salmo salar L.) smolts at constant pH in bicarbonate rich freshwater. Aquaculture 178:171–187CrossRefGoogle Scholar
  25. Fivelstad S, Olsen AB, Åsgård T, Baeverfjord G, Rasmussen T, Vindheim T, Stefansson S (2003) Long-term sublethal effects of carbon dioxide on Atlantic salmon smolts (Salmo salar L.): ion regulation, haematology, element composition, nephrocalcinosis and growth parameters. Aquaculture 215:301–319CrossRefGoogle Scholar
  26. Gaulke GL, Dennis CE III, Wahl DH, Suski CD (2014) Acclimation to a low oxygen environment alters the hematology of largemouth bass (Micropterus salmoides). Fish Physiol Biochem 40:129–140CrossRefPubMedGoogle Scholar
  27. Gregory TR, Wood CM (1998) Individual variation and interrelationships between swimming performance, growth rate, and feeding in juvenile rainbow trout (Oncorhynchus mykiss). Can J Fish Aquat Sci 55:1583–1590CrossRefGoogle Scholar
  28. Hartley HO (1950) The maximum F-ratio as a short-cut test for heterogeneity of variance. Biometrika 37:308–312PubMedGoogle Scholar
  29. Hasler CT, Butman D, Jeffrey JD, Suski CD (2016) Freshwater biota and rising pCO2? Ecol Lett 19:98–108CrossRefPubMedGoogle Scholar
  30. Healy TM, Tymchuk WE, Osborne EJ, Schulte PM (2010) Heat shock response of killifish (Fundulus heteroclitus): candidate gene and heterologous microarray approaches. Physiol Genom 41:171–184CrossRefGoogle Scholar
  31. Heuer RM, Grosell M (2014) Physiological impacts of elevated carbon dioxide and ocean acidification on fish. Am J Physiol 307:R1061–R1084Google Scholar
  32. Iwama GK, Heisler N (1991) Effect of environmental water salinity on acid-base regulation during environmental hypercapnia in the rainbow trout (Oncorhynchus mykiss). J Exp Biol 158:1–18Google Scholar
  33. Iwama GK, McGeer JC, Pawluk MP (1989) The effects of five fish anaesthetics on acid–base balance, hematocrit, blood gases, cortisol, and adrenaline in rainbow trout. Can J Zool 67:2065–2073CrossRefGoogle Scholar
  34. Iwama GK, Afonso LO, Todgham A, Ackerman P, Nakano K (2004) Are hsps suitable for indicating stressed states in fish? J Exp Biol 207:15–19CrossRefPubMedGoogle Scholar
  35. Kassahn KS, Crozier RH, Pörtner HO, Caley MJ (2009) Animal performance and stress: responses and tolerance limits at different levels of biological organisation. Biol Rev 84:277–292CrossRefPubMedGoogle Scholar
  36. Kates D, Dennis C, Noatch MR, Suski CD, MacLatchy D (2012) Responses of native and invasive fishes to carbon dioxide: potential for a nonphysical barrier to fish dispersal. Can J Fish Aquat Sci 69:1748–1759CrossRefGoogle Scholar
  37. Kieffer J, Cooke S (2009) Physiology and organismal performance of centrarchids. In: Cooke SJ, Philipp DP (eds) Centrarchid fishes: diversity, biology, and conservation. Wiley, West Sussex, pp 207–263CrossRefGoogle Scholar
  38. Kristensen T, Åtland Å, Rosten T, Urke H, Rosseland B (2009) Important influent-water quality parameters at freshwater production sites in two salmon producing countries. Aquacult Eng 41:53–59CrossRefGoogle Scholar
  39. Leroi AM, Bennett AF, Lenski RE (1994) Temperature acclimation and competitive fitness: an experimental test of the beneficial acclimation assumption. Proc Natl Acad Sci USA 91:1917–1921CrossRefPubMedPubMedCentralGoogle Scholar
  40. Lewis JM, Hori TS, Rise ML, Walsh PJ, Currie S (2010) Transcriptome responses to heat stress in the nucleated red blood cells of the rainbow trout (Oncorhynchus mykiss). Physiol Genom 42:361–373CrossRefGoogle Scholar
  41. Logan CA, Somero GN (2011) Effects of thermal acclimation on transcriptional responses to acute heat stress in the eurythermal fish Gillichthys mirabilis (Cooper). Am J Physiol Regul Integr Comp Physiol 300:R1373–R1383CrossRefPubMedGoogle Scholar
  42. Lowry O, Passonneau J (1972) A flexible system of enzymatic analysis. Academic, New YorkGoogle Scholar
  43. Lurman GJ, Bock CH, Poertner H (2009) Thermal acclimation to 4 or 10 C imparts minimal benefit on swimming performance in Atlantic cod (Gadus morhua L.). J Comp Physiol B 179:623–633CrossRefPubMedGoogle Scholar
  44. Mancebo MJ, Ceballos FC, Pérez-Maceira J, Aldegunde M (2013) Hypothalamic neuropeptide Y (NPY) gene expression is not affected by central serotonin in the rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol A 166:186–190CrossRefGoogle Scholar
  45. McEwen BS, Wingfield JC (2003) The concept of allostasis in biology and biomedicine. Horm Behav 43:2–15CrossRefPubMedGoogle Scholar
  46. McKenzie DJ, Piccolella M, Dalla Valle AZ, Taylor EW, Bolis CL, Steffensen JF (2003) Tolerance of chronic hypercapnia by the European eel Anguilla anguilla. J Exp Biol 206:1717–1726CrossRefPubMedGoogle Scholar
  47. Melzner F, Göbel S, Langenbuch M, Gutowska MA, Pörtner H, Lucassen M (2009) Swimming performance in Atlantic Cod (Gadus morhua) following long-term (4–12 months) acclimation to elevated seawater PCO2. Aquat Toxicol 92:30–37CrossRefPubMedGoogle Scholar
  48. Michaelidis B, Spring A, Pörtner HO (2007) Effects of long-term acclimation to environmental hypercapnia on extracellular acid–base status and metabolic capacity in Mediterranean fish Sparus aurata. Mar Biol 150:1417–1429CrossRefGoogle Scholar
  49. Munday PL, Dixson DL, Donelson JM, Jones GP, Pratchett MS, Devitsina GV, Doving KB (2009) Ocean acidification impairs olfactory discrimination and homing ability of a marine fish. Proc Natl Acad Sci USA 106:1848–1852CrossRefPubMedPubMedCentralGoogle Scholar
  50. Munday PL, Dixson DL, McCormick MI, Meekan M, Ferrari MC, Chivers DP (2010) Replenishment of fish populations is threatened by ocean acidification. Proc Natl Acad Sci USA 107:12930–12934CrossRefPubMedPubMedCentralGoogle Scholar
  51. Niu CJ, Rummer JL, Brauner C, Schulte PM (2008) Heat shock protein (Hsp70) induced by a mild heat shock slightly moderates plasma osmolarity increases upon salinity transfer in rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol C 148:437–444Google Scholar
  52. Perry S, Gilmour K (2006) Acid–base balance and CO2 excretion in fish: unanswered questions and emerging models. Resp Physiol Neurobiol 154:199–215CrossRefGoogle Scholar
  53. Petochi T, Di Marco P, Priori A, Finoia M, Mercatali I, Marino G (2011) Coping strategy and stress response of European sea bass Dicentrarchus labrax to acute and chronic environmental hypercapnia under hyperoxic conditions. Aquaculture 315:312–320CrossRefGoogle Scholar
  54. Piersma T, Drent J (2003) Phenotypic flexibility and the evolution of organismal design. Trends Ecol Evol 18:228–233CrossRefGoogle Scholar
  55. Raven J, Caldeira K, Elderfield H, Hoegh-Guldberg O, Liss P, Riebesell U, Shepherd J, Turley C, Watson A (2005) Ocean acidification due to increasing atmospheric carbon dioxide. The Royal Society. The Clyvedon Press Ltd., CardiffGoogle Scholar
  56. Raymond PA, Hartmann J, Lauerwald R, Sobek S, McDonald C, Hoover M, Butman D, Striegl R, Mayorga E, Humborg C (2013) Global carbon dioxide emissions from inland waters. Nature 503:355–359CrossRefPubMedGoogle Scholar
  57. Reidy S, Nelson J, Tang Y, Kerr S (1995) Post-exercise metabolic rate in Atlantic cod and its dependence upon the method of exhaustion. J Fish Biol 47:377–386CrossRefGoogle Scholar
  58. Riebesell U, Fabry VJ, Hansson L, Gattuso J (2010) Guide to best practices for ocean acidification research and data reporting. Publications Office of the European Union LuxembourgGoogle Scholar
  59. Rimoldi S, Terova G, Brambilla F, Bernardini G, Gornati R, Saroglia M (2009) Molecular characterization and expression analysis of Na+/H+ exchanger (NHE)-1 and c-Fos genes in sea bass (Dicentrarchus labrax, L) exposed to acute and chronic hypercapnia. J Exp Mar Biol Ecol 375:32–40CrossRefGoogle Scholar
  60. Robbins L, Hansen M, Kleypas J, Meylan S (2010) CO2calc—a user-friendly seawater carbon calculator for Windows, Max OS X, and iOS (iPhone). US Geol Surv, Open-File Rep 1280 Google Scholar
  61. Ross RM, Krise WF, Redell LA, Bennett RM (2001) Effects of dissolved carbon dioxide on the physiology and behavior of fish in artificial streams. Environ Toxicol 16:84–95CrossRefPubMedGoogle Scholar
  62. Santos G, Schrama J, Capelle J, Rombout J, Verreth J (2013) Effects of dissolved carbon dioxide on energy metabolism and stress responses in European seabass (Dicentrarchus labrax). Aquacult Res 44:1370–1382CrossRefGoogle Scholar
  63. Serrano X, Grosell M, Serafy J (2010) Salinity selection and preference of the grey snapper Lutjanus griseus: field and laboratory observations. J Fish Biol 76:1592–1608CrossRefPubMedGoogle Scholar
  64. Simpson SD, Munday PL, Wittenrich ML, Manassa R, Dixson DL, Gagliano M, Yan HY (2011) Ocean acidification erodes crucial auditory behaviour in a marine fish. Biol Lett 7:917–920CrossRefPubMedPubMedCentralGoogle Scholar
  65. Sobek S, Tranvik LJ, Cole JJ (2005) Temperature independence of carbon dioxide supersaturation in global lakes. Glob Biogeochem Cy 19:GB2003CrossRefGoogle Scholar
  66. Sokal RR, Rohlf FJ (1995) Biometry. Freeman and Company, New YorkGoogle Scholar
  67. Suski CD, Cooke SJ, Tufts BL (2007) Failure of low velocity swimming to enhance recovery from exhaustive exercise in largemouth bass (Micropterus salmoides). Phys Biochem Zool 80:78–87CrossRefGoogle Scholar
  68. Tankersley CG, Haxhiu MA, Gauda EB (2002) Differential CO2-induced c-fos gene expression in the nucleus tractus solitarii of inbred mouse strains. J Appl Physiol 92:1277–1284CrossRefPubMedGoogle Scholar
  69. Telmer K, Veizer J (1999) Carbon fluxes, pCO2 and substrate weathering in a large northern river basin, Canada: carbon isotope perspectives. Chem Geol 159(1–4):61–86CrossRefGoogle Scholar
  70. Thomsen J, Gutowska M, Saphörster J, Heinemann A, Trübenbach K, Fietzke J, Hiebenthal C, Eisenhauer A, Körtzinger A, Wahl M (2010) Calcifying invertebrates succeed in a naturally CO2-rich coastal habitat but are threatened by high levels of future acidification. Biogeosciences 7:3879–3891CrossRefGoogle Scholar
  71. US Department of the Interior, US Fish and Wildlife Service, and US Department of Commerce, US Census Bureau (2011) 2011 National survey of fishing, hunting, and wildlife-associated recreation. Washington, DCGoogle Scholar
  72. Wells RM (2009) Blood-gas transport and hemoglobin function: adaptations for functional and environmental hypoxia. Fish Physiol 27:255–299CrossRefGoogle Scholar
  73. Wendelaar Bonga SE (1997) The stress response in fish. Physiol Rev 77:591–625PubMedGoogle Scholar
  74. Zar JH (1984) Biostatistical analysis. Prentice Hall, USAGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Clark E. DennisIII
    • 1
  • Shivani Adhikari
    • 1
  • Adam W. Wright
    • 1
  • Cory D. Suski
    • 1
    Email author
  1. 1.Department of Natural Resources and Environmental SciencesUniversity of IllinoisUrbanaUSA

Personalised recommendations