tion, some commonly used oligo- and polysaccharides were included in the study to permit ... dextran and xylan (Fluka), glycogen and cellulose. (Merck), kappa ... by digestion of starch, but free maltose may be a minor constituent of the food.
Offprint from "Marine Biology" International Journal on Life in Oceans and Coastal Waters, Vol. 14, No.2, May 1972, Pages 130-142
© by Springer- Verlag 1972 . Printed in Germany
Carbohydrases of some marine invertebrates with notes on their food and on the natural occurrence of the carbohydrates studied J.
HYLLEBERG KRISTENSEN
Zoological Institute, Laboratorium B, Ecology, University of Aarhus; Aarhus, Denmark
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HYLLEBERG KRISTENSEN:
Carbohydrases of marine invertebrates
Abstract Extracts of 22 ma·rine invertebrates were incubated with 29 different carbohydrates, and enzyme spectra estimated from chromatograms. Enzymatic activities were quantified as reducing sugar in 14 of these species. Significant hydrolysis of laminaran, glycogen, and amylose was found in nearly all species, while hydrolysis of oligosaccharides and structural polysaccharides in most cases was weak or absent. The strongest overall degradation of carbohydrates was found among crustaceans. Enzymatic degradation of laminaran shows some relation to food, but the spectra of carbohydrases are not directly predictable from knowledge of natural food sources. Some carbohydrases (sucrase, cellobiase, lactase) could not be explained by ecological considerations. The results indicate that structural polysaccharides are utilized only to a small extent.
Introduction
FENCHEL (1970) observed that bacteria and other microflora were removed from detrital particles during their passage through amphipods, while the detritus itself remained undigested. This observation, and studies by CROSSLEY et ai. (1963), GEORGE (1964), NEWELL (1965), HARGRAVE (1970), and KrSSELEVA and VITYUK (1970), indicate that detritus, defined as dead organic matter, plays a minor role in the nutrition of detritus feeders. This paper presents the in vitro activity of carbohydrases extracted from benthic animals mainly ingesting detritus composed of inert substances (Fox, 1950) such as structural carbohydrates. The enzymatic splitting of cellulose, hemicellulose and algal poly-uronic acids was, therefore, especially studied. An attempt is made to estimate the significance of these substances as energy sources of detritus feeders by comparing the activity of carbohydrases. In addition, some commonly used oligo- and polysaccharides were included in the study to permit comparison with more readily hydrolysed substrates. An increasing interest in carbohydrases during the last decades has resulted in a number of papers presenting different approaches. Comparative studies on degradation of carbohydrates have been carried out on groups of ecologically comparable species. NIELSEN (1962) made a qualitative comparison of
Ma'f. Bioi.
carbohydrases in 34 soil and litter invertebrates, and B. O. NIELSEN (1966) in 6 wrack invertebrates. The present work is, to my knowledge, the first of this kind which compares carbohydrases in marine invertebrates, many of which have common feeding habits. Other papers deal with the spectrum of carbohydrases of single taxonomical groups or of species; these studies deal chiefly with the quantitative aspect of enzymatic hydrolysis. Degradation has also been investigated in selected animals with respect to particular carbohydrates. Many papers deal with cellulases (references by YOKOE and YASUMASU, 1964) chitinases (extensively treated by JENIAUX, 1963), laminaranases (SOVA et aI., 1970), and alginases (FRANSSEN and JENIAUX, 1965; FAVAROV and VASKOVSKY, 1971). Materials and methods
For the qualitative studies, 19 detritus feeders and omnivorous species were collected during spring and early summer from the estuarine Niva Bay (Muus, 1967), i.e., the bivalves Macoma balthica (L.), Mya arenaria L., Mytilus edulis L., and Cardium edule L., the gastropods Hydrobia ulvae (PENNANT), H. ventrosa (MONTAGU), and Littorina littorea L., the amphipods Gammarus zaddachi SEXTON, Bathyporeia pilosa LINDSTR0M, B. sarsi WATKIN and C01'Ophium volutator (PALLAS), the decapods Carcinus maenas L., and Crangon crangon (FAllR.), the isopod Idothea balthica (PALLAS), the barnacle Balanus crenatus BRUG., the polychaetes Nereis divel'sicolor O. F. MULLER, Pygospio elegans CLAPAR:EJDE, Arenicola marina L., and the oligochaete Paranais littoralis MULLER. Three species, the gastropod Nassarius reticulatus (L.) the sea star Asterias rubens L. and the brittle star Ophiocomina nigra ABILDG., were collected in the 0resund and used for comparative purposes. They were fed mussel meat and kept for varying periods of time in aquaria. The other species were dissected shortly after collection, and digestive organs (Fig. 1) were ground at 6°C with HCI-washed quartz. Some small species (Fig. 1) were washed in buffer and homogenized in toto.
Vol. 14, No.2, 1972
J.
HYLLEBERG KRISTENSEN:
Carbohydrases of marine invertebrates
Samples were diluted to ca. 1: 25 with citric acidphosphate buffer, pH 5.6. Some extracts were also made at pH 7.6. (Hydrobia ulvae H. ventrosa, Nassarius reticulatus and Asterias rubens) and pH 4.3. (N. reticulatus, Ophiocomina nigra and A. rubens). After centrifugation, the supernatant was used as enzyme preparation and 20 fLI added to each of the following 29 commercial carbohydrates (about 0.5 mg of each): the U' and G. J. DODSON: GroSs effects of arthropods and microBora on rates of leaf litter breakdown. In: Progress in terrestrial and fresh water ecology, pp 98-99. Health Physics Div. A. Prog. Rep. ORNL - 3492, U.S. Atomic Energy Commission 1963. DUCKWORTH, M. and J. R. TuRVEY: An extracellular agarase from a Cytophaga species. Biochem. J. 113, 139-142 (1969). EpPLEY, R. W. and R. LASKER: Alginase in the sea urchin Strongylocentrotus purpuratus. Science, N.Y. 129, 214-215 (1959). FAVOROV, V. V. and V. E. VASKOVSKY: Alginases of marine invertebrates. Comp. Biochem. Physio!. 38, 689-696 (1971). FENCHEL, T.: Studies on the decomposition of organic detritus derived from the turtle grass Thala8sia testudinum. Lim· no!. Oceanogr. 16, 14-20 (1970). FmQUET, M.-P. et G. DANDRIFOSSE: Influence du regime alimentaire sur les proprietes catalytiques de l'amylase pancreatique. Arcbs into Physio!. Biochim. 78, 347-355 (1967). FONTAINE, A. R.: The feeding mechanism of the ophiuroid Ophiowmina nigra. J. mar. bio!' Ass. U.K. 46, 373-385 (1965). FOSTER, A. B. and M. STACEY: The polysaccharides from lower plants such as"bacteria"algae, fungi and lichens etc. In: Encyclopedia of plant physiology,"Vo!. 6. pp 331334. Ed. by W. RUHLAND. Springer Verlag: Berlin 1958. Fox, D. L.: Comparative metabolism of organic betritus by inshore animals. Ecology 31,100-108 (1950). FRANSSEN, J. et CR. JENIAUX: Digestion de l'acide alginique chez les invertebres. Cah. Bio!. mar. 6, 1-21 (1965). GALLI, D. R. and A. C. GmsE: Carbohydrate digestion in a herbivorous snail Tegula lumbralis. J. expo Zool. 140, 415-440 (1959).
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