Marine Biology

, Volume 149, Issue 4, pp 865–873 | Cite as

Digestive enzymes in the ontogenetic stages of the southern king crab, Lithodes santolla

  • R. Saborowski
  • S. Thatje
  • J. A. Calcagno
  • G. A. Lovrich
  • K. Anger
Research Article

Abstract

The early ontogenetic stages of the sub-Antarctic king crab Lithodes santolla were analyzed for the presence and activities of a set of important digestive enzymes. The eggs and non-feeding larvae (zoea I-III, megalopa) showed high activities of esterases, phosphatases, and exopeptidases indicating the enzymatic ability to utilize endogeneous yolk reserves. SDS-PAGE showed a continuous decrease of proteins or proteids in the range of 59–81 kDa during ontogenetic development from the eggs through the zoeal stages to the first juvenile crab stage, CI. This reduction reflects the degradation of storage compounds during lecithotrophic larval development. Activities of the endopeptidases, trypsin and chymotrypsin, were low in eggs and larvae but increased significantly in the first juvenile crab stage. These enzymes typically facilitate the first steps of proteolysis in the extra-cellular spaces of the midgut gland and in the stomach. Their scarcity indicates that the larvae of L. santolla are physiologically not prepared to digest external food. This ability seems to appear first in the CI stage. Extracts of juvenile midgut glands and the gastric fluids of adults showed high activities of a variety of digestive enzymes including phosphatases, carbohydrases, as well as endo- and exopeptidases. High activities of digestive enzymes in adults may compensate for scarce food supply and rate-limiting low temperatures in the predominantly sub-Antarctic habitats of L. santolla.

References

  1. Anger K (2001) The biology of decapod crustacean larvae. Crustacean issues 14. AA Balkema Publishers, Swets and Zeitlinger, Lisse, 420 ppGoogle Scholar
  2. Anger K, Lovrich G, Thatje S, Calcagno J (2004) Larval and early juvenile development of Lithodes santolla (Molina, 1782) (Decapoda: Anomura: Lithodidae) reared at different temperatures in the laboratory. J Exp Mar Biol Ecol 306:217–230CrossRefGoogle Scholar
  3. Balzi PP (1997) Feeding habits of southern king crab, Lithodes santolla (Molina), in the San Jorge Gulf. Naturalia Patagonica Cienc Biol 5:67–87Google Scholar
  4. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  5. Calcagno JA, Anger K, Lovrich GA, Thatje S, Kaffenberger A (2004) Larval development of the subantarctic king crabs Lithodes santolla and Paralomis granulosa reared in the laboratory. Helgol Mar Res 58:11–14CrossRefGoogle Scholar
  6. Carnevali O, Mosconi G, Cambi A, Ridolfi S, Zanuy S, Polzenetti-Magni AM (2001) Changes of lysosomal enzyme activities in sea bass (Dicentrarchus labrax) eggs and developing embryos. Aquaculture 202:249–256CrossRefGoogle Scholar
  7. Chen L, Jiang H, Zhou Z, Li K, Li K, Deng GY, Liu Z (2004) Purification of vitellin from the ovary of Chinese mitten-handed crab (Eriocheir sinensis) and development of an antivitellin ELISA. Comp Biochem Physiol B 138:305–311PubMedCrossRefGoogle Scholar
  8. Comoglio L, Amin O (1996) Natural diet of the southern king crab Lithodes santolla in the Beagle Channel, Tierra del Fuego, Argentina. Biol Pesq 25:51–57Google Scholar
  9. Dall W, Moriarty DJW (1983) Functional aspects of feeding and digestion. In: Mantel LH (ed) The biology of crustacea. Vol 5. Internal anatomy and physiological regulation. Academic, New York, pp 215–261Google Scholar
  10. Dawson EW (1989) King crabs of the world (Crustacea: Lithodidae) and their fisheries: a comprehensive bibliography. Misc Publ, vol 101. New Zealand Oceanogr Inst, Div Water Sci, DSIR, WellingtonGoogle Scholar
  11. Fagotto F (1995) Regulation of yolk degradation, or how to make sleepy lysosomes. J Cell Sci 108:3645–3647PubMedGoogle Scholar
  12. Fang LS, Lee BN (1992) Ontogenetic change of digestive enzymes in Penaeus monodon. Comp Biochem Physiol 103B:1033–1037Google Scholar
  13. Galgani F, Nagayama F (1987) Digestive proteases in five species of Lithodidae (Crustacea, Decapoda). Comp Biochem Physiol B 87:103–107CrossRefGoogle Scholar
  14. Garçia-Carreño FL, Dimes LE, Haard NF (1993) Substrate-gel electrophoresis for composition and molecular weight of proteinases or proteinaceous proteinase inhibitors. Anal Biochem 214:65–69PubMedCrossRefGoogle Scholar
  15. Geiger R (1988) Chymotrypsin. In: Bergmeyer HU (ed) Methods of enzymatic analysis. 3rd edn, vol 5. Enzymes 3: peptidases, proteinases and their inhibitors. VCH Verlagsgesellschaft, Weinheim, pp 99–118Google Scholar
  16. Geiger R, Fritz H (1988) Trypsin. In: Bergmeyer HU (ed) Methods of enzymatic analysis. 3rd edn, vol 5. Enzymes 3: peptidases, proteinases and their inhibitors. VCH Verlagsgesellschaft, Weinheim, pp 119–129Google Scholar
  17. Hafkenscheid JCM (1988) Amino acid arylamidase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. 3rd edn, vol 5. Enzymes 3: peptidases, proteinases and their inhibitors. VCH Verlagsgesellschaft, Weinheim, pp 11–15Google Scholar
  18. Jones DA, Kumlu M, Le Vay L, Fletcher DJ (1997) The digestive physiology of herbivorous, omnivorous and carnivorous crustacean larvae: a review. Aquaculture 155:289–299CrossRefGoogle Scholar
  19. Kattner G, Graeve M, Calcagno JA, Lovrich GA, Thatje S, Anger K (2003) Lipid, fatty acid and protein utilization during lecithotrophic larval development of Lithodes santolla (Molina) and Paralomis granulosa (Jacquinot). J Exp Mar Biol Ecol 292:61–74CrossRefGoogle Scholar
  20. Kumlu M, Jones DA (1997) Digestive protease activity in planktonic crustaceans feeding at different trophic levels. J Mar Biol Assoc UK 77:159–165CrossRefGoogle Scholar
  21. Lemos D, Hernández-Cortés MP, Navarrete A, Garcia-Carreño FL, Phan VN (1999) Ontogenetic variation in digestive proteinase activity of larvae and postlarvae of the shrimp Farfantepenaeus paulensis (Crustacea: Decapoda: Penaeidae). Mar Biol 135:653–662CrossRefGoogle Scholar
  22. Lemos D, Garcia-Carreño FL, Hernández P, Navarrete del Toro MA (2002) Ontogenetic variation in digestive proteinase activity, RNA and DNA content of larval and postlarval white shrimp Litopenaeus schmitti. Aquaculture 214:363–380CrossRefGoogle Scholar
  23. Le Moullac G, Klein B, Sellos D, Van Wormhoudt A (1996) Adaptation of trypsin, chymotrypsin and α-amylase to casein level and protein source in Penaeus vannamei (Crustacea Decapoda). J Exp Mar Biol Ecol 208:107–125CrossRefGoogle Scholar
  24. Le Vay L, Jones DA, Puello-Cruz AC, Sangha RS, Ngamphongsai C (2001) Digestion in relation to feeding strategies exhibited by crustacean larvae. Comp Biochem Physiol A 128:623–660Google Scholar
  25. Lovrich GA, Thatje S, Calcagno JA, Anger K, Kaffenberger A (2003) Changes in biomass and chemical composition of the southern king crab, Lithodes santolla (Molina). J Exp Mar Biol Ecol 288:65–79CrossRefGoogle Scholar
  26. McLaughlin PA, Anger K, Kaffenberger A, Lovrich GA (2001) Megalopal and early juvenile development in Lithodes santolla (Molina, 1782) (Decapoda, Anomura; Paguroidea: Lithodidae), with notes on zoeal variations. Invertebr Reprod Dev 40:53–67Google Scholar
  27. McLaughlin PA, Anger K, Kaffenberger A, Lovrich GA (2003) Larval and early juvenile development in Paralomis granulosa (Jacquinot) (Decapoda: Anomura: Paguroidea: Lithodidae), with emphasis on abdominal changes in megalopal and crab stages. J Nat Hist 37:1433–1452CrossRefGoogle Scholar
  28. Saborowski R, Sahling G, Navarrete del Toro MA, Walter I, García-Carreño FL (2004) Stability and effects of organic solvents on endopeptidases from the gastric fluid of the marine crab Cancer pagurus. J Mol Catal B Enzym 30:109–118CrossRefGoogle Scholar
  29. Serrano-Pinto V, Vazquez-Boucard C, Villarreal-Colmenares H (2003) Yolk proteins during ovary and egg development of mature female freshwater crayfish (Cherax quadricarinatus). Comp Biochem Physiol A 134:33–43CrossRefGoogle Scholar
  30. Thatje S, Anger K, Calcagno JA, Lovrich GA, Pörtner HO, Arntz WE (2005) Challenging the cold: crabs reconquer the Antarctic. Ecology 86(3):619–625Google Scholar
  31. Thorson G (1936) The larval development, growth and metabolism of Arctic marine bottom invertebrates compared with those of other seas. Medd Grnland 100:1–155Google Scholar
  32. Thorson G (1950) Reproductive and larval ecology of marine bottom invertebrates. Biol Rev 25:1–45CrossRefGoogle Scholar
  33. Tom M, Fingerman M, Hayes TK, Johnson V, Kerner B, Lubzens E (1992) A comparative study of the ovarian proteins from two penaeid shrimps, Penaeus semisulcatus (de Haan) and Penaeus vannamei (Bone). Comp Biochem Physiol B 102:483–490CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • R. Saborowski
    • 1
  • S. Thatje
    • 2
    • 3
  • J. A. Calcagno
    • 4
  • G. A. Lovrich
    • 5
  • K. Anger
    • 1
  1. 1.Biologische Anstalt HelgolandFoundation Alfred Wegener Institute for Polar and Marine ResearchHelgolandGermany
  2. 2.Alfred Wegener Institute for Polar and Marine ResearchBremerhavenGermany
  3. 3.National Oceanography Centre, SouthamptonUniversity of Southampton SouthamptonUK
  4. 4.Facultad de Sciencias Exactas y NaturalesUniversidad Buenos Aires, Consejo Nacional de Investigaciones Científicas y TécnicasBuenos AiresArgentina
  5. 5.Consejo Nacional de Investigaciones Científicas y TécnicasCentro Ausral de Investigaciones Científicas, CADIC, CC 92UshuaiaArgentina

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