Journal of Comparative Physiology B

, Volume 183, Issue 6, pp 749–761 | Cite as

Physiological and biochemical responses to cold and drought in the rock-dwelling pulmonate snail, Chondrina avenacea

  • Vladimír KoštálEmail author
  • Jan Rozsypal
  • Pavel Pech
  • Helena Zahradníčková
  • Petr Šimek
Original Paper


The pulmonate snail Chondrina avenacea lives on exposed rock walls where it experiences drastic daily and seasonal fluctuations of abiotic conditions and food availability. We found that tolerance to dry conditions was maintained at a very high level throughout the year and was mainly based on the snails’ ability to promptly enter into estivation (quiescence) whenever they experienced drying out of their environment. Snails rapidly suppressed their metabolism and minimized their water loss using discontinuous gas exchange pattern. The metabolic suppression probably included periods of tissue hypoxia and anaerobism as indicated by accumulation of typical end products of anaerobic metabolism: lactate, alanine and succinate. Though the drought-induced metabolic suppression was sufficient to stimulate moderate increase of supercooling capacity, the seasonally highest levels of supercooling capacity and the highest tolerance to subzero temperatures were tightly linked to hibernation (diapause). Hibernating snails did not survive freezing of their body fluids and instead relied on supercooling strategy which allowed them to survive when air temperatures dropped to as low as −21 °C. No accumulation of low-molecular weight compounds (potential cryoprotectants) was detected in hibernating snails except for small amounts of the end products of anaerobic metabolism.


Mollusca Estivation Hibernation Metabolic suppression Water loss Supercooling 



Relative humidity of the air


Supercooling point


Total fresh mass


Total dry mass


Shell mass


Body water mass


Body dry mass


Time of exposure lethal to 50 % of a population sample


Discontinuous gas exchange


Principal component analysis



We thank the Administration of Protected Landscape Area Český kras for permitting us to sample snails in the study site of Solvayovy lomy. Lucie Juřičková (Charles University, Department of Zoology, Prague) recommended to us the locality with dense population of the snails. Roman Hrdlička, Jaroslav Šoun and Hana Pechová helped during field sampling. We thank Irena Vacková, Anna Heydová and Jana Cimlová (all from the Biology Centre, ASCR, České Budějovice) for their assistance with snail sample processing, extractions, derivatizations and LC/MS analysis, respectively. This study was supported by Czech Science Foundation grant 206/07/0269 (to VK), Ministry of Health of the Czech Republic grant NT-11/513-5 (to PŠ), and University of South Bohemia grant GAJU 04-062/2011/P (to JR).

Supplementary material

360_2013_749_MOESM1_ESM.docx (1.9 mb)
Supplementary material 1 (DOCX 1919 kb)
360_2013_749_MOESM2_ESM.docx (28 kb)
Supplementary material 2 (DOCX 29 kb)
360_2013_749_MOESM3_ESM.docx (18 kb)
Supplementary material 3 (DOCX 19 kb)


  1. Ansart A, Vernon P (2003) Cold hardiness in molluscs. Acta Oecol 24:95–102CrossRefGoogle Scholar
  2. Ansart A, Vernon P (2004) Cold hardiness abilities vary with the size of the land snail Cornu aspersum. Comp Biochem Physiol A 139:205–211CrossRefGoogle Scholar
  3. Ansart A, Vernon P, Daguzan J (2001a) Photoperiod is the main cue that triggers supercooling ability in the land snail, Helix aspersa (Gastropoda: Helicidae). Cryobiol 42:266–273CrossRefGoogle Scholar
  4. Ansart A, Vernon P, Daguzan J (2001b) Freezing tolerance versus freezing susceptibility in the land snail Helix aspersa (Gastropoda: Helicidae). Cryo-Lett 22:183–190Google Scholar
  5. Ansart A, Vernon P, Daguzan J (2002) Elements of cold hardiness in a littoral population of the land snail Helix aspersa (Gastropoda: Pulmonata). J Comp Physiol B 172:619–625PubMedCrossRefGoogle Scholar
  6. Ansart A, Nicolai A, Vernon P, Madec L (2010) Do ice nucleating agents limit the supercooling ability of the land snail Cornu aspersum? Cryo-Lett 31:329–340Google Scholar
  7. Attia J (2004) Behavioural rhythms of land snails in the field. Biol Rhythm Res 35:35–41CrossRefGoogle Scholar
  8. Bailey SER (1981) Circannual circadian rhythms in the snail (Helix aspersa Müller) and the photoperiodic control of annual activity and reproduction. J Comp Physiol 142:89–94CrossRefGoogle Scholar
  9. Barnhart MC (1983) Gas permeability of the epiphragm of a terrestrial snail, Otala lactea. Physiol Zool 56:436–444Google Scholar
  10. Barnhart MC (1986a) Respiratory gas tensions and gas exchanges in active and dormant land snails, Otala lactea. Physiol Zool 59:733–745Google Scholar
  11. Barnhart MC (1986b) Control of acid-base status in active and dormant land snails, Otala lactea (Pulmonata, Helicidae). J Comp Physiol B 156:347–354CrossRefGoogle Scholar
  12. Barnhart MC, McMahon BR (1987) Discontinuous carbon dioxide release and metabolic depression in dormant land snails. J Exp Biol 128:123–138Google Scholar
  13. Barnhart MC, McMahon BR (1988) Depression of aerobic metabolism and intracellular pH by hypercapnia in land snails, Otala lactea. J Exp Biol 138:289–299Google Scholar
  14. Baur A, Baur B (1991) The effect of hibernation position on winter survival of the rock-dwelling land snails Chondrina clienta and Balea perversa on Öland, Sweden. J Mollusc Stud 57:331–336CrossRefGoogle Scholar
  15. Biannic M, Daguzan J (1993) Cold-hardiness and freezing in the land snail Helix aspersa Muller (Gastropoda, Pulmonata). Comp Biochem Physiol A 10:503–506CrossRefGoogle Scholar
  16. Bueding E, Orrell SA (1964) A mild procedure for the isolation of polydisperse glycogen from animal tissue. J Biol Chem 239:4018–4020PubMedGoogle Scholar
  17. Caputa M, Nowakowska A, Rogalska J, Wentowska K (2005) Winter torpor in Helix pomatia: regulated defence mechanism or forced inactivity? CanJ Zool 83:1608–1613CrossRefGoogle Scholar
  18. Chown SL, Nicolson SW (2004) Insect physiological ecology. Oxford, New YorkCrossRefGoogle Scholar
  19. Churchill TA, Storey KB (1989) Intermediary energy metabolism during dormancy and anoxia in the land snail Otala lactea. Physiol Zool 62:1015–1030Google Scholar
  20. Denlinger DL (1991) Relationships between cold hardiness and diapause. In: Lee RE Jr, Denlinger DL (eds) Insects at low temperature. Chapman and Hall, New York, pp 174–198CrossRefGoogle Scholar
  21. DuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  22. Elmslie LJ (2001) Strain differences in Helix pomatia L. (Gastropoda: Pulmonata): diapause, digging, growth rate and shell fill. J Mollusc Stud 67:121–124CrossRefGoogle Scholar
  23. Gessner MO, Neumann PTM (2005) Total lipids. In: Graça MAS, Bärlocher F, Gessner MO (eds) Methods to study litter decomposition: a practical guide. Springer, Berlin, pp 103–107Google Scholar
  24. Giokas S, Pafilis P, Valakos E (2005) Ecological and physiological adaptations of the land snail Albinaria caerulea (Pulmonata: Clausiliidae). J Mollus Stud 71:15–23CrossRefGoogle Scholar
  25. Guppy M, Fuery CJ, Flanigan JE (1994) Biochemical principles of metabolic depression. Comp Biochem Physiol B 109:175–189PubMedGoogle Scholar
  26. Hermes-Lima M, Storey JM, Storey KB (1998) Antioxidant defenses and metabolic depression. The hypothesis of preparation for oxidative stress in land snails. Comp Biochem Physiol B 120:437–448PubMedCrossRefGoogle Scholar
  27. Herreid CF (1977) Metabolism of land snails (Otala lactea) during dormancy arousal and activity. Comp Biochem Physiol A 56:211–215PubMedCrossRefGoogle Scholar
  28. Hoback WW, Stanley DW (2001) Insects in hypoxia. J Insect Physiol 47:533–542PubMedCrossRefGoogle Scholar
  29. Hochachka PW, Lutz PL (2001) Mechanism, origin, and evolution of anoxia tolerance in animals. Comp Biochem Physiol B 130:435–459PubMedCrossRefGoogle Scholar
  30. Kerney MP, Cameron RAD, Jungbluth JH (1983) Die Landschnecken Nord- und Mitteleuropas. Verlag Paul Parey, Hamburg und BerlinGoogle Scholar
  31. Kokshoorn B, van Schoor M, Erkelens I, Gittenberger E (2010) Waves of dispersal in island-hopping Chondrina species (Gastropoda, Pulmonata, Chondrinidae). Zoologischer Anzeiger 249:71–79CrossRefGoogle Scholar
  32. Koštál V (2006) Eco-physiological phases of insect diapause. J Insect Physiol 52:113–127PubMedCrossRefGoogle Scholar
  33. Koštál V, Berková P, Šimek P (2003) Remodelling of membrane phospholipids during transition to diapause and cold-acclimation in the larvae of Chymomyza costata (Drosophilidae). Comp Biochem Physiol B 135:407–419PubMedCrossRefGoogle Scholar
  34. Koštál V, Korbelová J, Rozsypal J, Zahradníčková H, Cimlová J, Tomčala A, Šimek P (2011) Long-term cold acclimation extends survival time at 0 °C and modifies the metabolomic profiles of the larvae of the fruit fly Drosophila melanogaster. PLoS ONE 6:e25025PubMedCrossRefGoogle Scholar
  35. Lind H (1989) Homing to hibernating sites in Helix pomatia involving detailed long term memory. Ethology 81:221–234CrossRefGoogle Scholar
  36. Machin J (1967) Structural adaptation for reducing water loss in three species of terrestrial snail. J Zool (London) 152:55–65CrossRefGoogle Scholar
  37. Machin J (1968) The permeability of the epiphragm of terrestrial snails to water vapor. Biol Bull 134:87–95CrossRefGoogle Scholar
  38. Machin J (1972) Water exchange in the mantle of a terrestrial snail during periods of reduced evaporative loss. J Exp Biol 57:103–111Google Scholar
  39. Michaelidis B (2002) Studies on the extra- and intracellular acid–base status and its role on metabolic depression in the land snail Helix lucorum (L.) during estivation. J Comp Physiol B 172:347–354PubMedCrossRefGoogle Scholar
  40. Newell PF, Machin J (1976) Water regulation in aestivating snails. Ultrastructural and analytical evidence for an unusual cellular phenomenon. Cell Tissue Res 173:417–421PubMedCrossRefGoogle Scholar
  41. Nicolai A, Vernon P, Lee M, Ansart A, Charrier M (2005) Supercooling ability in two populations of the land snail Helix pomatia (Gastropoda: Helicidae) and ice-nucleating activity of gut bacteria. Cryobiol 50:48–57CrossRefGoogle Scholar
  42. Nicolai A, Filser J, Lenz R, Bertrand C, Charrier M (2012) Quantitative assessment of hemolymph metabolites in two physiological states and two populations of the land snail Helix pomatia. Physiol Biochem Zool 85:274–284PubMedCrossRefGoogle Scholar
  43. Nopp H (1974) Physiologische Aspekte des Trockenschlafes der Landschnecken. Sitzungsberichte-Oesterreichishe Akad Wiss. Math-Naturwiss Klasse 182:1–75Google Scholar
  44. Nowakowska A, Caputa M, Rogalska J (2006) Seasonal changes in cryoprotectants concentrations in Helix pomatia snails. J Physiol Pharmacol 57:93–105PubMedGoogle Scholar
  45. Nowakowska A, Swiderska-Kolacz G, Rogalska J, Caputa M (2009) Antioxidants and oxidative stress in Helix pomatia snails during estivation. Comp Biochem Physiol C 150:481–485Google Scholar
  46. Nowakowska A, Caputa M, Rogalska J (2011a) Defence against oxidative stress in two species of land snails (Helix pomatia and Helix aspersa) subjected to estivation. J Exp Zool 315:593–601CrossRefGoogle Scholar
  47. Nowakowska A, Caputa M, Rogalska J (2011b) Effects of temperature and photoperiod on glucose, glycerol and glycogen concentrations in Helix pomatia Linnaeus 1758 in spring and autumn. Fol Malacol 19:155–163CrossRefGoogle Scholar
  48. Oosterhoff LM (1977) Variation in growth rate as an ecological factor in the land snail Cepaea nemoralis. Neth J Zool 27:1–132CrossRefGoogle Scholar
  49. Pedler S, Fuery CJ, Withers PC, Flanigan J, Guppy M (1996) Effectors of metabolic depression in an estivating snail (Helix aspersa): whole animal and in vitro studies. J Comp Physiol B 166(375):381Google Scholar
  50. Pollard D (1975) Aspects of the ecology of Helix pomatia L. J Anim Ecol 44:305–329CrossRefGoogle Scholar
  51. Ramos-Vasconcelos GR, Cardoso LA, Hermes-Lima M (2005) Seasonal modulation of free radical metabolism in estivating land snails Helix aspersa. Comp Biochem Physiol C 140:165–174CrossRefGoogle Scholar
  52. Rees BB, Hand SC (1990) Heat dissipation, gas exchange and acid-base status in the land snail Oreohelis during short-term estivation. J Exp Biol 152:77–92Google Scholar
  53. Riddle WA (1981) Cold hardiness in the woodland snail Anguispira alternata (say) (Endodontidae). J Therm Biol 6:117–120CrossRefGoogle Scholar
  54. Riddle WA, Miller VJ (1988) Cold-hardiness in several species of land snails. J Therm Biol 13:163–167CrossRefGoogle Scholar
  55. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH et al (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85PubMedCrossRefGoogle Scholar
  56. Storey KB (1977) Lactate dehydrogenase in tissue extracts of the land snail, Helix aspersa: unique adaptation of LDH in a facultative anaerobe. Comp Biochem Physiol B 56:181–186PubMedGoogle Scholar
  57. Storey KB (2002) Life in the slow lane: molecular mechanisms of estivation. Comp Biochem Physiol A 133:733–754CrossRefGoogle Scholar
  58. Storey KB, Storey JM (1990) Metabolic rate depression and biochemical adaptation in anaerobiosis, hibernation and estivation. Quart Rev Biol 65:145–174PubMedCrossRefGoogle Scholar
  59. Storey KB, Storey JM (1991) Biochemistry of cryoprotectants. In: Lee RE Jr, Denlinger DL (eds) Insects at low temperature. Chapman and Hall, New York, pp 64–93CrossRefGoogle Scholar
  60. Stöver H (1973) Cold-resistance and freezing in Arianta arborustorum L. (Pulmonata). In: Wieser W (ed) Effects of temperature on ectothermic animals. Springer, Berlin, pp 281–290CrossRefGoogle Scholar
  61. Ultsch GR, Carwile ME, Crocker CE, Jackson DC (1999) The physiology of hibernation among turtles: the eastern painted turtle, Chrysemys picta picta. Physiol Zool 72:493–501CrossRefGoogle Scholar
  62. Zachariassen KE (1985) Physiology of cold tolerance in insects. Physiol Rev 65:799–832PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Vladimír Koštál
    • 1
    • 2
    Email author
  • Jan Rozsypal
    • 1
    • 2
  • Pavel Pech
    • 3
  • Helena Zahradníčková
    • 1
  • Petr Šimek
    • 1
  1. 1.Institute of EntomologyBiology Centre of the Academy of SciencesČeské BudějoviceCzech Republic
  2. 2.Faculty of ScienceUniversity of South BohemiaČeské BudějoviceCzech Republic
  3. 3.Faculty of ScienceUniversity of Hradec KrálovéHradec KrálovéCzech Republic

Personalised recommendations