Sterols in Marine Invertebrates

FISHERIES SCIENCE 2001; 67: 997–1007

Review Article

Sterols in marine invertebrates

A KANAZAWA*

Emeritus Professor, Faculty of Fisheries, Kagoshima University, Kagoshima 890-0057, Japan

ABSTRACT: The sterol composition of marine invertebrates, crustaceans, molluscs, echinoderms, coelenterates, and sponges has been re-examined by using improved analytical techniques. As a result, the sterol composition of marine invertebrates has been shown to be more complex mixtures including many new types of sterols with unusual steroid nuclei or with non-conventional side chains. Some crustaceans and molluscs are of importance as seafood and aquaculture species. Crustaceans and some molluscs require dietary sources of sterol for growth and survival because of the absence of de novo sterol-synthesizing ability. The present paper gives a review of the requirement and nutritive value of sterols, biosynthesis and metabolism of sterols, and composition and structure of sterols in marine invertebrates.

KEY WORDS: biosynthesis, crustacean, invertebrate, mollusc, sterol.

INTRODUCTION thesize sterol. Such studies have employed [14C]- labeled acetate or mevalonate injected into the The sterol constituents of a variety of marine animal followed by incubations ranging from a few invertebrates have been reinvestigated in detail by hours to several days before isolation of sterols for using modern techniques such as gas–liquid chro- radioassay. Part of the results is summarized in matography, argentation chromatography, mass Table1.1 spectrometry, and nuclear magnetic resonance spectrometry. On the other hand, several workers have investigated the sterol biosynthesis in marine Crustaceans invertebrates. The lack of sterol synthesis in some marine invertebrates is of interest in the viewpoint Mammals can synthesize cholesterol from low of both chemotaxonomy and animal nutrition. molecular weight precursors such as acetate or Some crustaceans and molluscs in marine inverte- mevalonate. However, a unique aspect of the lipid brates also include important species for aquacul- nutrition of crustaceans and some molluscs is ture. The present paper presents the nutritive that they require dietary sources of sterol for value, metabolism, and composition of sterols in growth and survival because of the absence of marine invertebrates. These topics have been de novo sterol-synthesizing ability.7,16 In higher described in reviews by many researchers during animals, cholesterol is an important precursor the past two decades.1–15 of steroid hormones, molting hormones, bile salts, and vitamin D. It has been demonstrated in spiny lobsters that exogenous cholesterol is con- BIOSYNTHESIS AND REQUIREMENT verted to sex hormones such as pregesterone, OF STEROLS 17a-hydroxyprogesterone, androstenedione, and testosterone, and to molting hormones such Many investigations have been conducted to as 20-hydroxyecdysone.17 A feeding experiment determine the capacity of representative species of using artiﬁcial diets conducted by Kanazawa8 ﬁrst all the major marine invertebrate phyla to biosyn- demonstrated that Penaeus japonicus requires sterols for growth and survival, and reported a value of 0.5% dietary cholesterol for good *Corresponding author: Tel: 099-251-6272. Fax: 099-251-6352. growth. The optimum or required levels of dietary Received 15 January 2001. Accepted 31 May 2001. cholesterol for crustaceans reported to date are 998 FISHERIES SCIENCE A Kanazawa

Table 1 Summary of the evidence for, or against, de shrimp. On the other hand, it suggested that novo sterol biosynthesis in marine invertebrate animals. banana shrimp may require lecithin between 1 and 1 (Goad ) 2% in the diet for good growth and high survival. Phylum Class Sterol However, there was no advantage given by supp- biosynthesis* lementing cholesterol to the basal diet. [4-14C]- Positive Negative Cholesterol was orally administered to P.japonicus Porifera 6 5 to clarify the effects of dietary phospholipids on Cnidaria Anthozoa 5 7 the mobilization of sterols in diets to various 37 Scyphozoa – 1 organs and tissues. The results suggest that Nemertini 2 – dietary phospholipid contributes to the smooth Annelida Polychaeta 7 – mobilization of dietary cholesterol in the body, Arthropoda Crustacea – 16 especially from the hepatopancreas to the Mollusca Amphineura 3 – hemolymph. On the other hand, Baum et al.38 have Gastropoda 27 2 shown that lobsters fed diets supplemented with Bivalvia 6 11 lecithin had significantly higher levels of serum Cephalopoda 2 4 and lipoprotein cholesterol than lobsters fed diets Scaphopoda – 1 without supplemental lecithin. Levels of fecal Echinodermata Crinoidea 1 – Asteroidea 8 – total cholesterol also were generally greater for Holothuroidea 7 1 lobsters fed lecithin-supplemented diets. In 1997, 28 Echinoidea 7 1 Teshima et al. have indicated that the juvenile Ophiuroidea 3 – Macrobrachium rosenbergii was capable of de novo Urochordata 4 – cholesterol synthesis in contrast to other prawn species but required a dietary source of 0.1% cho- *Table indicates the reported results obtained from the lesterol for the maximum growth. Briggs et al.27 incubation of marine animals with radioactive acetate or mevalonate. found that a level of 0.12% cholesterol was suffi- cient for juvenile M. rosenbergii. Using a semi- purified diet, D’Abramo and Daniels39 found a significant increase in weight gain of the juvenile summarized to be approximately 0.1–2.0% of the M. rosenbergii when the level of dietary cholesterol dry weight of diet (Table2), and may be age and was increased from 0.3 to 0.6%, but a further diet dependent. In the larval prawn, P. japonicus, increase to 0.9 and 1.2% produced no higher the effectiveness of dietary soybean lecithin in weight gain. In the absence of a dietary source of improving growth and survival has not been sterol, M. rosenbergii died within 48 days after the shown to be affected by dietary cholesterol levels, start of the experiment. Recently, D’Abramo29 has indicating that the optimum soybean lecithin reviewed the nutritional requirements of M. rosen- and cholesterol levels were 3.0% and 1.0% in the bergii and compared them with those reported for diets, respectively.21 A growth study using juvenile several species of marine shrimp. A combination of Penaeus monodon fed test diets containing graded dietary phytosterols is equally effective as choles- levels of cholesterol (0, 0.5 and 1% of diet) and terol in satisfying a dietary sterol requirement of purified phosphatidylcholine (0, 1.25, 2.5 and 5% 0.6% for juvenile M. rosenbergii. Penaeid species of diet) in combination showed that the shrimp require sterols, but dietary phytosterols are not attained optimal growth when diets contained 1% as effective as cholesterol in promoting growth. cholesterol or 1.25% phosphatidylcholine.23 The Perhaps M. rosenbergii has the ability to synthesize values are equal to those obtained for Penaeus cholesterol from dietary b-sitosterol at a more effi- penicillatus.24 The interactions between choles- cient rate. Crustaceans possess the ability to terol and phosphatidylcholine on weight gain of dealkylate some C28 and C29 sterols to cholesterol. both species were insignificant. Recently, Thon- Kanazawa et al.40 and Teshima and Kanazawa41 grod and Boonyaratpalin25 have determined have demonstrated that survival rates of juvenile cholesterol and lecithin requirements of juvenile prawns receiving ergosterol, stigmasterol, or b- banana shrimp, Penaeus merguiensis. The choles- sitosterol were high, but growth was inferior when terol feeding experiment showed that shrimp fed compared with that of prawns receiving choles- the diet without cholesterol supplement had terol as a sterol source. In addition, prawn larvae similar weight gain to shrimp fed a diet supple- have also been shown to grow well with high sur- mented with 0.5 or 1% cholesterol. This indicates vival rates on the microparticulate diets containing that the non-cholesterol supplemented diet that 22-dehydrocholesterol, 24-methylenecholesterol, already contained 0.6% sterol esters, might have ergosterol, or isofucosterol.42 These suggest the satisfied the cholesterol requirement of banana possible C-24 dealkylation of these C28 and C29 Sterols in marine invertebrates FISHERIES SCIENCE 999

Table 2 Cholesterol requirements of crustaceans Species Optimum sterol Researcher (% in diets) Prawn: Penaeus japonicus 0.5–1.0 Kanazawa et al. (1971)18 0.2 Shudo et al. (1971)19 2.1 Deshimaru & Kuroki (1974)20 1.0 Teshima et al. (1982)21 P chinensis 1.0 Kanazawa et al. (unpubl. data) P monodon 0.54 Kai & Kanazawa (1989)22 0.5 Chen (1993)23 P penicillatus 0.5–1.0 Chen & Jenn (1991)24 P merguiensis 0.6 Thongrod & Boonyaratpalin(1998)25 Argentine prawn: Artemesia longinaris 0.5 Petriella et al. (1984)26 Freshwater prawn: Macrobrachium rosenbergii 0.12 Briggs et al. (1988)27 0.11–0.26 Teshima et al. (1997)28 0.6 D’Abramo (1998)29 Lobster: Homarus americanus 0.5 Castell et al. (1975)30 Non-essential (Adult) Castell & Covey (1976)31 0.12 D’Abramo et al. (1984)32 0.19 0.25 Kean et al. (1985)33 0.2 Bordner et al. (1986)34 Crayﬁsh: Pacifastacus leniusculus 0.4 D’Abramo et al. (1985)35 Crab: Carcinus maenas 1.4–2.1 Ponat & Adelung (1983)36

sterols to cholesterol in crustaceans. Teshima43 squid, Sepia officinalis.46 However, no incorpora- proposed a pathway of dealkylation of C28 and C29 tion of acetate and mevalonate into sterols has sterols via desmosterol to cholesterol in crus- been shown in the mussel, M. edulis,47 the oyster, taceans (Fig. 1). Ostrea gryphea,48 and the cephalopods, Eledon aldovadi.46 Generally, pelecypods and cephalopods seem to incorporate acetate and mevalonate into Molluscs sterols poorly. Accordingly, pelecypods are likely to require dietary sources of sterols for growth and The sterol metabolism of molluscs differs between survival. Trider and Castell49 have suggested, by classes.43 Chitons belonging to class Amphineura feeding experiments using artificial diets, that the can synthesize cholest-7-enol from low molecular oyster, Crassostrea virginica, requires low levels of weight precursors and also transform exogenous cholesterol (0.1–0.2%) for growth. However, the test cholesterol to cholest-7-enol. Many gastropod diets used did not show a significant increase in species have been shown to possess the ability for oyster tissue weight. It should be noted that trials de novo synthesis of cholesterol. In general, cho- of the oyster were conducted using seawater fil- lesterol biosynthesis seems to proceed at a rapid tered down to 10 mm; therefore, the sterol require- rate in gastropods. Gastropods are also probably ments of oysters and other pelecypods should be capable of dealkylating some phytosterols to cho- re-evaluated more rigorously in consideration of lesterol. It is possible that most gastropods do not microbial contribution. Teshima and Patterson50 require a dietary source of cholesterol for growth. also showed biosynthesis of 24-methylene- There is evidence for de novo sterol biosynthesis cholesterol and 24-ethylidenecholesterol from in the pelecypods, mussels, Mytilus edulis,44,45 the acetate in the oyster, C. virginica. Crassostrea vir- cephalopods, octopus, Octopus vulgaris,45 and the ginica has a metabolic requirement for cholesterol 1000 FISHERIES SCIENCE A Kanazawa

Fig. 1 Proposed mechanism for

C-24 alkylation of C28 and C29 sterols in crustaceans (Teshima43).

but is unable to synthesize cholesterol from that, although sea scallops may be capable of sterol acetate. According to Wikfors et al.,51 most microal- biosynthesis, the incorporation of unmodified gae including species that support rapid growth of dietary phytosterols plays an influential role in oyster, contained very little cholesterol, but pro- establishing their sterol composition. In 1999, duced a number of other sterols. Sixteen algal Knauer et al.53 have shown the assimilation of strains were evaluated for sterol composition semi- dietary phytosterols by Pacific oyster, Crassostrea quantitatively. Two aspects of algal sterol structure: gigas spat. Pacific oyster spat were fed either the (i) the presence of an ethyl rather than a methyl microalga, Chaetoceros muelleri, Isochrysis aff. group on carbon 24; and (ii) the presence of a galbana or Pavlova lutheri to investigate the effect double bond at carbon 5, showed statistically sig- of dietary phytosterols on the sterol composition of nificant positive effects upon oyster growth. The spat tissues. After a 6-week feeding period the anatomical distributions of sterols and the incor- sterol profile of spat tissues generally reflected that poration of dietary phytosterols were studied in of the diet. 4-Demethylsterols, such as cholesterol sea scallops, Placopecten magellanicus.52 In order and 24-methylcholestrol, were readily incorpo- to know if anisomyarian bivalves require a uniform rated into spat tissue but 4-methyl sterols were anatomical distribution of sterols, wild scallops poorly assimilated by spat. The absolute content of were subjected to a microalgal diet containing high every sterol decreased in spat that had been concentrations of brassicasterol, b-sitosterol and starved for 6 weeks, but the relative sterol compo- cholesterol. The sterol composition of the scallop sition was similar to the initial sterol profile. adductor muscle was not changed by 6 weeks of However, the relative level of cholesterol increased, feeding the experimental diet. In contrast, the pro- indicating the important role of this sterol in spat portion of brassicasterol, b-sitosterol and choles- metabolism. terol in the digestive gland, and of brassicasterol and cholesterol in the male gonad, increased significantly. These results show that the typical even Echinoderms anatomical distribution of sterols of bivalves can be disrupted by a drastic change in diet and is The echinoderms belonging to the class Asteroidea therefore not subject to strict internal regulation. and Holothuroidea contain mainly D7-sterols. The Furthermore, the P. magellanicus results indicate sterols of these echinoderms are generally complex Sterols in marine invertebrates FISHERIES SCIENCE 1001

mixtures that are composed of C26-, C27-, C28- because the sterol biosynthesis in the sponges and C29-sterols. The biosynthesis of sterols from seemed not to take place or to proceed at a slow mevalonate in the class Asteroidea (starfish), rate. Therefore, the sterol components of sponges Leiaster leachii,54,55 Coronaster valsellatus,54 Pro- are likely to vary with habitat. Marsh et al.60 have toreaster nodsus,54 Henricia ohshimai,54 Coscinaste- shown dietary effects on oocyte yolk composition rias acutispina,56 Asterias rubens,57 Marthasterias in Capitella sp. I (Annelida: Polychaeta). Immature glacialis,57 Astropecten aurantianus,57 and Echi- Capitella sp. I was raised on four diets: Gerber naster sepositus,57 and class Holothuroidea, cereal, a commercial mixed grain cereal; TetraMin, Holothuria leucospilota,54 Cucumaria planci,58 Sti- a staple diet for aquarium fish comprising algal chopus regalis 58 was studied. The results indicated and animal tissues; Ulva sp.; and benthic diatoms. that all echinoderms are capable of synthesizing at After 2 weeks of culturing different populations least cholest-7-enol from acetate or mevalonate via of these diets, eggs were dissected from gravid probably squalene and lanosterol etc. However, females and analyzed for sterol composition. this animal seems not to possess the ability for The sterol profiles of all eggs were dominated by both alkylation at the C-24 position and the intro- cholesterol and cholest-5,24-dien-3b-ol (60%). A duction of a double bond at the C-22 position of principal component analysis of egg sterols dis- cholst-7-enol, therefore it may be assumed that in criminated between adult diets with cholesterol, the starfish C27-sterols such as cholest-7-enol are 23,24-dimethyl-cholesta-5-en-3b-ol, cholest-5,24- derived by both biosynthesis and exogenous dien-3b-ol and C29 sterol showing the greatest dif- sources but C26, C28, C29, and C30-sterols depend on ferences. diet only. Asteroids are also capable synthesizing cholest-7-enol from cholesterol via cholestanol. In coral reefs, Acanthaster planci takes the corals con- COMPOSITION OF STEROLS taining gorgosterol in the diet and can convert dietary 5-sterol into 7-sterol. Gorgostanol( 0) D D D The sterol composition occurring in various isolated from A. planci may be an intermediate in phyla of marine invertebrates has been reinvesti- the biosynthesis of acanthasterol( 7) from gorgos- D gated in the 1970s and 1980s. These sterols were terol( 5).59 D analyzed by thin-layer chromatography, column chromatography, high-performance liquid chromatography, gas–liquid chromatography (GLC), Coelenterates GLC-mass spectrometry, infrared absorption spectroscopy, mass spectrometry, and nuclear The biosynthesis of sterols from mevalonate in magnetic resonance spectrometry. As a result, the the coelenterates, Sarcophyta sp., Stereonephthya sterol composition of marine invertebrates has japonica, Acalycigorgia inermis, Ellisella rubra, been shown to be more complex. Dofleinia armata, and Parasicyonis actinostoloides has been demonstrated.54 It was shown that the four coelenterates, Sarcophyta sp., S. japonica, D. armata, and P.actinostoloides, are more or less Crustaceans capable of synthesizing sterols from mevalonate, but the two species of coelenterates, A. inermis and Cholesterol is the major sterol (more than 90–95% E. rubra, lack this ability. These results indicate that of total sterols) in crustaceans. Most crustaceans some coelenterates possess the sterol-synthesizing contain small amounts of C28 and C29 sterols in the ability from mevalonate but others do not. sterol mixtures. Pakrashi et al.61 have shown that the horseshoe crabs, Tachpleus gigas and Car- cinoscorpius rotundicauda, exceptionally contain

Porifera and annelids considerable amounts of C28 and C29 sterols such as campesterol (10%), stigmasterol (7%), b-sitosterol Walton and Pennock47 have investigated the (24%), and 28-isofucosterol (3%). Serrazanetti biosynthesis of sterol by porifera (sponges) and et al.62 showed the sterol constituents of isopod annelids. The sponges have been suspected to crustacean, Idotea balthica basteri. The main contain sterol mixtures including new types of component contained in the sterol fraction was sterols. The sterols with unusual steroid nuclei or cholesterol and the minor constituents were with non-conventional side chains have been iso- cholesta-5,22-dien-3b-ol, cholesta-5,24-dien- lated from the sponges. The sterols occurring in the 3b-ol, 24-methylenecholest-5-en-3b-ol, and 24- sponges are thought to be derived from exogenous ethylidenecholest-5-en-3b-ol. Fricke and Oehlen- sources or by the modiﬁcation of dietary sterols, schlaeger63 identiﬁed cholesterol, desmosterol, 1002 FISHERIES SCIENCE A Kanazawa

Fig. 2 Some new sterols from marine invertebrates.

and 22-dehydrocholesterol as the main sterol the complexity of dietary sterols arising through composition of the Antarctic amphipod, Themisto food chains.65–67 The structures of 21 sterols were gaudichaudii. established from pearl-oyster, Pinctada martensi. The major components were D5-sterols. It was especially characteristic of the pearl-oyster that Molluscs many of the saturated sterols and ring-saturated sterols were also present as lesser components.68 The sterol composition of molluscs varies from During the course of an extensive investigation class to class.7,64 Chitons, the most primitive mol- of mollusc sterols, several new sterols have been 7 luscs belonging to class Amphineura, contain D - isolated; C26-sterol (22E)-24-norcholesta-5,22- sterol, mainly cholest-7-enol, in contrast to other dien-3b-ol (1),69 (the number and structure of classes of molluscs which predominantly possess the sterols are shown in Fig. 2), C30-sterols (24Z)- D5-sterols. Gastropods and cephalopods contain 24-propyridene-cholest-5-en-3b-ol (2)70 and (24Z)- 71 primarily cholesterol with small amounts of C28 24-propyridenecholest-7-en-3b-ol (3), C26(or and C29 sterols. Pelecypods contain complex mix- C27)-norsterols, occelasterol [(22E,24S)-27-nor-24- 72 tures of C26, C27, C28, C29, and C30 sterols together methyl cholesta-5,22-dienol] (4) and patinosterol with cholesterol. More than 45 sterols were present [(22E,24S)-27-nor-24-methyl cholest-22-enol] (5)73 in the oyster, Crassostrea virginica, reﬂecting etc. Sterols in marine invertebrates FISHERIES SCIENCE 1003

Table 3 Sterol composition in Acanthaster planci and methylene-23,24-dimethylcholest-5-en-3b-ol] the relative retention times of gas–liquid chromatogra- (11),80 23-demethylgorgosterol [(22R,23R,24R)- 74 phy (GLC*)(Sato et al. ) 22,23-methylene-24-methylcholest-5-en-3b-ol] 81 82 Sterol acetate % RRT (12) and 25-hydroxy-24-methylcholesterol (13) etc. 5a-Cholestan-3b-ol 0.1 1.00 Cholesterol trace 1.00 5a-Cholest-7-en-3b-ol 6.2 1.16 (22E)-5 a-Ergosta-7,22-dien-3b-ol 8.7 1.32 Porifera (sponges) Ergost-5-en-3b-ol 2.9 1.32 5a-Ergost-7-en-3b-ol 70.0 1.56 The sponges have been suspected to contain 5a-Ergosta-7,24(28)-dien-3b-ol 1.4 1.56 sterol mixtures including new types of sterols of 23,24-Dimethyl-5 0.9 1.56 over 100.2,3,83 The characterization of sterols of a-cholesta-7,22-dien-3b-ol sponges has been intensively carried out by several 23-Demethylacanthasterol 0.6 2.04 groups of workers using modern techniques. As Gorgostanol 0.3 2.36 a result, sterols with unusual steroid nuclei or Gorgosterol trace 2.36 with non-conventional side chains have been Acanthasterol 8.9 2.75 isolated from sponges; for example, stanols with 84 *1.5% OV-17. one double bond at side chains, 26-methylated sterols such as aplysterol (14) and 24,28- didehydroaplysterol (15),85 19-nor-sterols (16) with

saturated and monosaturated C7, C8, C9, and C10 Echinoderms side chains,86 stanols with 3b-hydroxymethyl- 87,88 A-nor-5a-cholestane nucleus (17), and C29 It is generally recognized that asteroids and cyclopropene-containing sterol.89 After that, Doss holothurians usually contain D7-sterols, while and Djerassi90 isolated four new sterols with ophiuroids and echinoids contain D5-sterols. As an cyclopropene-containing side chains from the example, the sterol composition of asteroid, Acan- Caribbean sponge, Calyx podatypa. The major thaster planci is shown in Table3.74 Up to date, sterol was shown to be the 23-epimer of (23R)- some new sterols have been isolated from echino- 23H- isocalysterol (18). Gunasekera et al.91 showed derms: for example, acanthasterol[(22R,23R,24R)- a new sterol ester, 24,26-cyclo-5a-cholest-(22E)- 22,23-methylene-23,24-dimethyl-5a-cholest-7- en-3b-ol 4¢, 8¢, 12¢-trimethyl-tridecanoate from a en-3b-ol] (6),75 23-demethyl acanthasterol (7),74 deep water marine sponge, Xestospongia sp. gorgostanol[(22R,23R,24R)-22,23-methylene- Makarieva et al.92 isolated a novel sterol with an 23,24-dimethyl-5a-cholestan-3b-ol] (8),59 24- unusual side chain, 24-ethyl-26-norcholesta- methylcholesta-7,22,25-trien-3b-ol (9),76 and 5,22E,25-trien-3b-ol (baikalosterol) (19) from the amuresterol[22-trans-27-nor-(24S)-24-methylc- sponge, Baicalospongia bacilifera. The presence of holesta-7,22-dien-3b-ol] (10).77 Recently, sterols these non-conventional sterols in the sponges is from the sea cucumbers, Pseudostichopus trachus, not only interesting from the biochemical view- Holothuria nobilis, Holothuria scabra, Trochos- point, but also may provide useful data for the toma orientale, and Bathyplotes natans have been taxonomical classiﬁcation of sponges. isolated and studied. About 80 sterols were found and 69 of these were identiﬁed, including several new and rare ones.78 BIOACTIVE STEROLS

Estrogens and their derivatives are generally known Coelenterates to be effective hypocholesterolemic agents. However, clinical application is often restricted due Many investigations using modern techniques to their side-effects as sex hormones. Accordingly, have pointed out the complexity of sterols in coe- a number of workers have attempted to ﬁnd lenterates. Kanazawa et al.79 have shown the sterol hypocholesterolemic substances without hormone composition of 22 species of the coelenterates. The activity. Sterols isolated from marine organisms class Scyphozoa contained large portions of C27- have been shown to exert a hypocholesterolemic sterols such as cholesterol and 22-dehydrocholes- effect. The effect of 7-cholestenol (cholesta- terol. The class Anthozoa was rich in either 7-en-3b-ol) and 24-methylenecholesterol (24- 24-methylenecholesterol or 24-methylcholesterol. methylenecholesta-5-en-3b-ol) on the cholesterol Some new sterols have been isolated from the level in serum and liver of rats has been studied. 7- coelenterates: gorgosterol [(22R,23R,24R)-22,23- Cholestenol and 24-methylenecholesterol from 1004 FISHERIES SCIENCE A Kanazawa

molluscs were found to significantly decrease the 2 Djerassi C. Recent studies in the marine sterol field. Pure cholesterol level in both serum and liver of rat.93 Appl. Chem. 1981; 53: 873–890. Datta et al.94 have isolated a bioactive sterol from 3 Djerassi C. Marine sterol. In: Krogsgaard-Larsen P, Brøgger a sea pen, Pteroeides esperi. A cytotoxic sterol, Christensen S, Kofod H (eds). Natural Products and Drug cholesta-3 , 5 , 6 -triol (20) isolated from the sea Development. Alfred Benzon Symposium 20. Munksgaard, b a b Cophenhagen, 1984; 164–178. pen is the first report of its isolation from a natural 4 Kanazawa A. Nutrition of penaeid prawns and shrimps. source. The sterol compositions of several soft In: Taki Y, Primavera JH, Liobrera JA (eds). Proceedings of corals and gorgonians as well as the composition the 1st International Conference on Culture of Penaeid of their associated symbiotic dinoflagellates have Prawns/Shrimps. 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