FISHERIES RESEARCH BOARO OF CANADA

TECHNICAL REPORT NO. 334

1972

FISHERIES RESEARCH BOARD OF CANADA

Technical Reports

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FISHERIES RESEARCH BOARO OF CANAOA

TECHNICAL REPORT NO. 334

HFECT OF UNICELLULAR ALGAL LIPIDS ON OYSTER

LIPIDS AND WEIR FATrY ACID COMPOSITIONS

BY

TAKESHI WATANABE

Tokyo University of FisheriesMinato-ku

Tokyo. Japan

AND

R.G. ACKMAN

Department of the EnvironmentFisheries Research Board of Canada

Halifax LaboratoryHali fax, Nova Scotia

FISHERIES RESEARCH BOARD OF CANADAUalifax Laboratory. Halifax, N.S.

ABsrnACf

Lipids and fatty acids of American (Crassostrea virginica) and

European (Ostrea edulis) oysters were studied to determine the effects of fceding

on the unicellular algae Dunaliella tertiolecta. Changes during storage of C.

virginica for six months were also examined. Separate experiments were carried out

with the algae Isochcl}Sisgalbana and Dicrateria inornata and both species of oyster

but without prolonged storage. In addi tion to study of the total lipids the algal

lipids were separated into polar and non-polar fractions, and oyster lipids into

polar lipid and triglycerides as well 35 stery! esters. glyceryl ethers, and free

fatty acids where practical. The detailed fatty acid compositions show that major

fatty acids in the diet mostly have recognizable but limited effects on the oyster

lipid fatty acid composition, presumably because the oysters stabilize lipids at near

optimum composi tions. The two oyster species responded differently to dietary fatty

acids and/or different algae. Hitherto unknown C20

and C22 non-methylene-interrupted

fatty acids are included in the detailed fatty acid analyses.

-2-

INTRODUCTION

The relative nutritive value of different species of marine

phytoplankton for zooplankton and larger filter feeders has been the

subject of speculation and study by many authors. Raymont (1963)

has reviewed the results of studies on the feeding of copepods. and

Walne (l970b) has reported on the relative food value of nineteen

genera of phytoplankton for oyster larvae. Bardach (1968) has also

reviewed recent developments in artificial rearing of many types of

marine organisms. Much of the earlier work has been based on total

calories or on amino acid "nitrogen" • with a tendency to regard protein,

carbohydrates and lipids as gross nutrient classes. with Ii tUe attention

being devoted to "essential" components in detail, although Iwasaki et al.,

(1971) have given consideration to an "essential amino acid" index.

The lipid components of planktonic organisms are a major

fraction of the metabolic energy resources of the sea. Various studies

which have been carried out on the biochemical composition of planktonic

organisms have yielded valuable infomation on the amount of lipid, on

total lipid fatty acid composition. and on different lipid components

for both zoo- and phytoplankton (Ackman et al., 1964; 1968; Corner and

Cowey, 1968; Culkin and Morris, 1968; Raymont et al., 1968; Chuecas and

Riley, 1969). Recently, Lee et al., (1971) have examined the importance

of wax ester formation as well as details on fatty acids in other lipids

in the marine food chain step between phytoplankton and copepods.

- 3 -

Phytoplankton are now accepted as the primary sources

of certain "essential" fatty acids of marine invertebrates and fish

(Farkas et al., 1961; Kayama and TSllchiya, 1962; Kayama et al., 1963)

but the step between plant and primative filter-feeding animals has

received little attention. In particular the economic importance of

oysters and potential for aquaculture indicate a need for infomation

on the fatty cid composition of oysters fed specific algae. We have,

therefore. c tducted feeding experiments with oysters using pure cuI tUTed

unicellular algae to investigate the influence of algal lipid on oyster

lipid fatty acid composition.

The chief experimental animal used in our investigation was

the American oyster. Crossostrea virginica. It is plentiful in the

coastal waters of the United States from Texas to Maine, and also inhabits

various warmer and sheltered bays and estuaries in the maritime provinces

of Canada (Bousfield. 1960). The introduced European oyster. Ostrea edulis.

was used in a limited comparison study.

MATERIALS AND METHODS

All the feeding tests were carried out at the biological

facilities of the Fisheries Research Board of Canada at Ellerslie.

Prince Edward Island. The axenic algal cultures were grown at 20_2S o C

and under sunliJtht. An enriched seawater medium was used. Both American

oysters. C. virginica and European oysters.o. edulis used in experiments

I. II and III had been kept in filtered seawater for more than one year

at the b ologieal facilities. In these experiments oysters were fed

- 4 -

Dunaliella tertiolecta. In other experiments in which oysters were

fed Isochrysis galbana and Dicrateria inornata the test animals had

been held in natural conditions in the sea near Ellerslie, P. E. T.

prior to feeding.

In experiment I. the oysters were brought from cold water

(ca. 2°C) to room temperature (20 G e) and held for 1 day before the

feeding experiment. For experiments II and III the animals were

transferred from cold water about 2 hours before commencement of

feeding. A representative group of oysters from each lot was used as

an unfed control for each lot. One experimental technique was used forunialgal

all feedings. Four litres of I culture (in seawater) were placed in

a polyethylene tank (45 x 30 x 15 em) containing 10 oysters. The

temperature of the culture and of the seawater from which the oysters

were removed was 20_22°C. The oysters began to actively filter the

seawater within 5 minutes of being covered by the liquid. The cell

densi ty of the cultures in seawater was determined every 30 minutes by the

packed volume method. When the cell density dropped to about half of the

ini tial value (after 1 hour of feeding). the oysters were transferred

to a new lot of culture. The total active filtering time was 6 hr and from

2.2 to 6.3 g of algae were incorporated by oysters in the different

experimental groups (Tables 1 and 2). After feeding tests the experimental

and control oysters and samples of the algal cultures were placed in plastic

bags and cooled to ice-water temperature for transport to the Halifax

Laboratory of the Fisheries Research Board for analyses.

- 5 -

The American oysters (C. virginica) in experiments I and II were

respectively divided into two groups. One half of each group was

kept at 2°C for 6 months.

Cultures were harvested by gentle centrifugation and the pellet

of algal cells drained to determine approximate wet cell weights (Table I).

Subsequent operations were carried out under an atmosphere of nitrogen.

Methanol and/or chloroform were used to transfer the cells to separatory

funnels with water in limited amounts to maintain a monophasic solvent

for 15-30 min. Additional water was then added to arrive at the final

ratios of solvents indicated by Bligh and Dyer (1959). The bottom layer

was filtered and stripped for lipid recovery (Tables and II).

Extractions of oysters were carried out on total wet organic

tissues with chlorofonn:methanol (2: 1). Polar lipids and non-polar

lipids were separated on and recovered from a divinylbenzene copolymer

bead gel column (Sipos and Ackman, 1968). Non-polar lipids were then

separated into sterol esters. glycerol ethers, triglycerides, free fatty

acids (in the case of o. edulis), and sterols on silica gel TLC plates

(Pre coated Adsorbosi 1-5, Applied Science Laboratories, Inc.). Procedures

of lipid fractionation are presented schematically in Figure 1. For the

algal lipids, the non-polar fraction was directly saponified and

non-saponi fiables were extracted and weighed. The recovered fatty acids

were esterified with BF3

-MeOH reagent. Removal of non-saponifiable

materials from lipids, preparation of methyl esters (by BF3-MeOH

esterifications of acids, or by transesterification of triglyceride), and

all GLe operations (open-tubular columns coated with aDS), followed

procedures described previously (Sipos and Ackman, 1968; Ackman and

Hooper, 1970). Particular care was taken to avoid confusion between branched

acids and plasmalogen artifacts (Ackman, 1972), as oysters are rich in these

lip~ds (Sampugna et al •• 1972).

- 6 -

The principal aim of this program was to investigate the fate

of "essential" polyunsaturated fatty acids as they were ingested

naturally by oysters from algal sources. While this work was in progress

it was discovered in this laboratory that the eicosenoic and docosenoic

acids. which are broken down by open-tubular gas liquid chromatography

into constituent isomers. contained anomalous peaks which were in fact

non-methylene interrupted dienoic acids. These have been included in relevant

data from this studY,but since their evaluation is at an early stage

we prefer to include them in publications with more detailed discussion

of the results.

RESULTS AND DISCUSSION

Lipid levels in each fraction of samples analysed are shown

in Tables I and II. tn experiments I, II and III the low lipid recovery

in comparison with those of oysters later fed I. galbana and D. inornata,

probably reflects the extended holding period without food for these

animals (Ackman and Cormier (1967); Ackman et al., 1971)). The

percentages of polar lipids. relative to total lipid. recovered from

oysters in experiment I. II and III were found to be lower than those of

other experimental groups and that reported by Ackman et al., (1971),

possibly for the same reason. Even the lipid recovery of oysters

collected from natural conditions (1.59\ for C. virginica) were lower

than those reported by Ackman and Cormier (1967); Shimma and Taguchi (1964).

Sampugna et al., (1970) and others, but comparisons are difficult due to the

imponderables such as extraction technique, size, species, environment

and habitats (Ackman and Hingley, 1968). and the effects of seasonal

variations (Ackman and Cormier, 1967; Cerma et al., 1970; Stancher et al.,

1971). However. the substantial residue of triglyceride indicates that

- 7 -

the depletion of lipids is not especially abnonnal in view of seasonal

limitation on food supply and the extreme lowering of water temperature

in the winter in the original habitat in Prince Edward Island (which

is the northern limit for this species for this reason (Bousfield, 1960)).

The fatty acid patterns for lipids of D. tertiolecta (Table III) were found

to be very similar to the result reported by Ackman et al., (1968).

whereas the fatty acid composition of I. galbana (Table IV) differed

significantly in detail from the result obtained by DeMort (1970).

especially in the level of 22:6w3 relative to 20:Sw3. This presumably is

due to the difference of culture conditions such as nitrogen concentration,

light J temperature. culture age and particular strain (Otsuka, 1961;

Otsuka and Morimura, 1966; Ackman et al., 1964; 1968; Lee et al., 1971).

The two species of Haptophyceae, I. galbana and D. inornata, were also

found to be quite different in lipid composition. The influence of the unusually

higher cuI ture temperature and natural sunlight may be profound. Thin-

layer chromatography on silica gel revealed that the non-polar lipid

of I. galbana consisted mainly of sterols, a small amount of

diglyceride and several kinds of pigments. The latter apparently influenced

the high proportion of non-saponifiable material (73.96\), whereas TLC

of D. inornata total non-polar material (only 7.3\) showed sterol

esters, glycerol ethers, triglyceride and sterols as principal components.

One difference which we had hoped to exploit in fatty acid composition

between the two species was the higher proportion of 18:4w3, but not of

18:3w3, and of 20:Sw3, fatty acids in I. galbana. However, there was no

important quantitative difference between control and experimental oysters

for these acids as indicated by eLC of total lipids from oysters fed on

these two species of algae.

- 8-

The fatty acid composition obtained fOT each lipid class

from the oysters is presented in Tables V to VIn. The fatty acids

of polar fractions (presumably mostly phospholipids from chromatographic

behaviour; the actual proportion of phosphorus was not determined) from

experimental groups fed 3 species of algae showed no real differences

between experimental and control groups. although the polar lipid

fatty acids of C. virginica in experiment I. and of O. edulis, both

fed O. tertiolecta. showed an increase of 16:0 and 17:0 acids possibly

related to a decrease of polyunsaturated C20

and e22 acids. The latter are

slightly suspect as delays in analysis of lipids may have led to some

autoxidation. This result for phospholipids seems to be in agreement

with reports that the fatty acids are highly specific and do not vary

with dietary changes because of their role as important membrane

consti tuents (Carroll, 1965; Wolfe et al., 1965; Benson J 1966; Jezyk

and Penicnak, 1966; Keenan and Morrt~J 1970; Lee et al., 1971).

The fatty acids of triglyceride from oysters in experiment

showed di fferences in the proportions of C16

, C18

and C20 monoethy1enic

fatty acids between control and experimental groups. However, after 6

months storage the high proportion of these fatty acids in c. virginica

(reflecting the low amount of 16:0 acid in the initial sample of the

experimental group) tends to show a close parallel to those of control

oys ters and an increase of 20: 5003 and 22: 6003 acids corresponding to an

intake of high amounts of dietary 18: 3003 from D. tertiolecta. In

- 9-

experiment II, the proportion of these polyunsaturated acids does

not di ffer a great deal between the experimental and control groups

both in ioi tial and after experimental 6 months. However. a higher

amount of 20:5w3 and 22:6w3 than in experiment I was recognized in

ioi tial samples and increased after 6 months. These differences in

results obtained for experiments I and II are probably due to the

di fferences in experimental design as the test animals for I had been

allowed to adapt to feeding conditions before the start of the

experiment as mentioned above, while II test animals were warmed up

only an hour before feeding. In the initial analysis (not stored)

of c. virginiclJ of the other experimental groups respectively fed

I. galbana and D. inornata. the proportion of C16

, CIS' C20

and C22

polyunsaturated acids are relatively close to those of controls. In the

fatty acid composition of c. virginica a larger amount of 16:0 (34.8\

in control, 43.5\ in experimental fed I. galbana and 42.1\ in

experimental fed D. inornata) reflects lower levels of 20: 5w3 and

22:6w3 acids. However, in o. edulis somewhat lower levels of 16:0 acid.

accompanied by higher proportions of 20:5w3 and 22:6w3 acids. were found

in the triglycerides of oysters fed I. galbana. The accumulation of normal (20

and C22 polyunsaturated acids in the group fed I. galbana seems to

reflect directly incorporation of fatty acids from I. galbana, which

contain much more 20:5w3 and 22:6w3 than D. 1nornata. The same kind of

differences between C. virginica and o. edulis were also observed in

experiment III. In experiment III, a high proportion of 16:0 acid in

-10 -

O. edulis related to lower levels of polyunsaturated C20

and e22

acids

appears to correspond to low levels of normal dietary polyunsaturated

C20 and e22 3cids from D. tertiolecta. These results probably indicate

that the mode of fatty acid metabolism I especially in relation to the

incorporation of dietary fatty acid, is different between the two species,

C. virginica and o. edulis.

The striking features of the analyses of sterol esters in experiments

I. II and III. in which oysters were fed D. tertiolecta. are: an accumulation

of 14: 0 acid and the accompanying 4.8.) 2-trimethyl tridecanoic acid (cf. Ackman

et al. ,1971) and, reduction in levels of 18:0 acid. In the American oyster

c. virginica, fed I. galbana and D. inornata, increases of monoethylenic fatty

acids along with decreases of 15:0 and 16:0 acids are noticeable, especially

in the oysters fed D. inornata. A similar tendency was observed in o. edulis,

but the proportion of 14: 0 acid was found to be low in the group fed I. galbana.

Polyunsaturated C22 acids were absent in both groups, whether control or

experimental.

Our results show that the variations in fatty acid composition between

the major lipid classes (e.g. phospholipids and triglycerides) are to some

extent independent of food intake. The relative proportions of the important

classes of I ipids may therefore be partially responsible for the seasonal

variations in total fatty acids observed in many molluscs such as those

studied by Calzolari et al., (1971). The fatty acids in total lipid of the

brine shrimp Artemia salina also show some independence of specific phytoplankters

in the diet (Hinchcliffe and Riley, 1972). but different lipid classes were not

studied.

-11-

While this work was in progress it became apparent that the fatty

acid patterns for monoethylenic acids from c. virginica and other molluscs

differed significantly from those normally observed in marine oils and lipids

(for examples, see Ackman and Castell, 1966 and Addison et a1., 1972). A

partial elaboration of structures indicates that the unusual "monoethylenic"

components are eicosadienoic and docosadienoic acids, probably of 5,13 and 5, IS

unsaturation for the (20:2) acids and 7:15 and 1:17 unsaturates for the (22:2)

acids. The ( ) indicates that these do not belong to the usual methylene­

interrupted polyunsaturated families of fatty acids. The CI8 unknowns have

not been characterized at the time of writing. Figures 2 and 3 show the effect

of an isolation of "normal" monoethylenic C18 and C20 acids by argentative TLC,

while Figure 4 shows unusual configuratiof6 of the (22: 2) acids.

Because of the novelty of these results we have isolated the data for

the (20:2) and (22:2) acids in separate subtables V-B, VI-B and VIII-B in

addition to the complete tables and subtotal tables of the A series. There still

remain some C20 and C22 unknowns. but these apparently occur unsystematically.

It is remarkable that the (20:2) and (22:2) acids were not found in the o. edulis

fed Isochrysis and Dicrateria (compare Tables VII and VIII). but there is no

obvious explanation for this.

ACKNOWLEDGEMENT

The authors express their thanks to Mr. R. Drinnan for arranging

for cultures of the algae and for valuable advice. T: Watanabe was the

recipient of a National Research Council of Canada Post-doctoral Fellowship.

-12-

REFERENCES

Ackman, R.C.. 1972. Tn "Pro~Tess in the Chemistry of Fats and Other Lipids

ed. R.T. Holman. PeTRamon Press, New York, Vol. !I. pp. 165-297.

Ackman, R. G. and M. G. Comier. 1967. a-Tocopherol in some Atlantic

fish and shellfish with particular reference to live-holding

without food. J. Fish. Res. Bd. Canada ~J 357-373.

Ackman, R. G. and H. J. Hingley. 1968. The occurrence and retention of

dimethyl-Bpropiothetin in some filter-feeding organisms.

J. Fish. Res. Bd. Canada ~. 267-284.

Ackman. R. G. and S. N. Hooper. 1970. Branched-chain fatty acids of

freshwater fish oil. Compo Biochem. Physiol. g. 117-125.

Ackman, R. G.• S. N. Hooper and P. J. Ke. 1971. The distribution of

saturated and isoprenoid fatty acids in the lipids of three

species of molluscs, Littorina littorea, Crassostrea virginica

and Venus mercenaria. Comp. Biochem. Physio1. ~, 579-587.

Ackman, R. G.• P. M. Jangaard, R. J. Hoyle and H. Brockerhoff. 1964.

Origin of marine fatty acids. I. Analyses of the fatty acids

produced by the Diatom Skeletonemo costatum. J. Fish. Res. Bd.

Canada ~, 747-756.

Ackman, R. G., C. S. Tocher and J. McLachlan. 1968. Marine phytop1ankter

fatty acids. J. Fish. Res. Bd. Canada ~, 1603-1620.

Addison, R.F .• R.G. Ackman and J. Hing1ey. 1972. Lipid composition of the

queen crab (Chionoecetes opi.lio). J. Fish. Res. Bd. Canada 29: 407-411.

Bardach, J. E. 1968. Aquaculture. Science .!!!., 1098-1106.

Benson, A. A. 1966. On the orientation of lipids in ch1oropast and cell

membranes. J. Am. Oil Chem. Soc. Q, 265-270.

-13-

Bligh, E. G. and W. J. Dyer. 1959. A rapid method of total lipid

extraction and purification. Can. J. Biochcm. Physiol. lZ..

911-917.

Bousfield, E. L. 1960. Canadian Atlantic Sea Shells. National Museum

of Canada. 72 pages.

Calzolari, C., E. Cerma, and B. Stancher. 1971. Gas chromatography applied

in determining fatty acids of some gastropoda and lalllcllibranchia from

Adriatic sea during an annual cycle. Riv. Ital. Sost. Grasse XLVI II,

605-616.

Carroll, K. K. 1965. Dietary fat and fatty acid composition of tissue

lipids. J. Am. Oil Chern. Soc. £. 516-552.

Cerma, E.• B. Stancher and P. Baradel. 1970. Molluscs of the upper

Adriatic Sea (Chemical composi tian of some of the Gastropods and

the Lamellibrachs). Rassegna Chimica (Chemical Review), ~.

39-43 & 3 unpaged sheets.

Chuecas. L. and J. P. Riley. 1969. Component fatty acids of the total

lipid of some marine phytoplankton. J. Mar. 8iol. Ass. U. K.

~, 97-116.

Corner, E. D. S. and C. B. Cowey. 1968. Biochemical studies on the

production of marine zooplankton. BioI. Rev. ~. 399-426.

Culkin, F. and R. J. Morris. 1969. The fatty acids of some marine

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DeMort, C. L. 1910. The culture and biochemical analysis of some estuarine

phytoplankton species. A thesis for the degree of Doctor of

Philosophy in Oregon State Universi ty. 157 pages.

Farkas, T.• S. Herodek, L. CS'aki and G. Toth. 1961. Incorpor3tion of

acetate_I_C I4 into the liver fatty acids of the fish Amiurus

nebulosus. Acta BioI. Acad. Sci. Hung. g. 83-86.

Hinchcliffe, P.R., and J.P. Riley. 1972. The effect of diet on the component

fatty acid composition of Artemla salina . .J. mar. biol. Ass. U.K.

-14-

Iwasaki J H., S. Tanaka and T. Fujiyama. 1971. On Rhodomonas ovalis

Nygaard as food for clam larvae. Bull. Jap. Soc. Sci. Fish.

Vol. 37. 1044-1048.

Jezyk. P. F. and A. J. Penicnak. 1966. Fatty acid relationships in an

aquatic food chain. Lipids!.. 427-429.

Kayama. M. and Y. Tsuchiya. 1962. Possible conversion pathway of

polyunsaturated acid in fish. Tohoku J. Agriculture Res. !l.

229-235.

Kayama, M.• Y. Tsuchiya and J. F. Mead. 1963. A model experiment of

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conversion. Bull. Jap. Soc. Sci. Fish. ~J 452-458.

Keenan, T. W. and O. J. MarrEL 1970. Phospholipid class and fatty acid

composi tian of golgi apparatus isolated from rat Iiver and

comparison wi th other cell fractions. Biochemistry U.S. A.

~. 19-25.

Lee J R. F .• J. C. Nevenzel and G. A. Paffenhttfer. 1971. Importance of

wax esters and other lipids in the marine food chain:

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Otsuka. H. 1961. Changes of lipid and carbohydrate contents in Chlorella

cells during the sulfur starvation as studied by the technique

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Otsuka. H. and Y. Morimura. 1966. Changes of fatty acid composition of

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Raymont, J. 1963. Plankton and productivity in the oceans. New York,

Pergamon. 660 p.

-15-

Raymont, J.E.G., R.T. Srinivasagam and J.K.B. Raymont. 1969. Biochemical

studi es on marine zooplankton. VI. Investigation of Heganyctiphanes

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Sampugna, J .• L. Johnson, K. Bachman and M. Kenney. 1972. Lipids of

Crassostrea virginica. 1. Prel iminary investigations of aldehyde and

phosphorous containing lipids in oyster tissue. Lipids 2.. 339-343.

Sampugna. J., M. Keeney, K. Bachmann and L. Johnson. 1970. Lipids from

the oyster Crassostrea virginica. Unpublished data. Am.

Oil Che•. Soc. (Abstract froID. 2nd Int. Soc. World Fat Res.

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Shimma. Y. and H. Taguchi. 1964. A comparative study of fatty acid

composition of shellfish. Bull. Jap. Soc. Sci. Fish. ~' 153-160.

Sipos, J. C. and R. G. Ackman. 1968. Jellyfish (Cyanea capillata) lipids:

Fatty acid composition. J. Fish. Res. Bd. Canada ~' 1561-1569.

Stancher J B., E. Cerma and P. Baradel. 1971. Mollusks of the upper

Adriatic. Variations of the chemical composi tion of some

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chimica (Chemical Review), ~' 39-42.

Wa1ne, P. R. 1970a. The seasonal variation of meat and glycogen content

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-16-

Wolfe. D. A., P. V. Rao and D. G. Cornwell. 19-5. The fatty acid

composition of crayfish lipids. J. Am. Oil Chern. Soc. £.

633-637.

Gel column

Total lipid

Saponi fication

IPolar lipid

INon polar lipid

ITotal fatty acids

INon saponi fiable

Fatty acids Non sap.

Methyl esters uu.. GLC

Hydrogenated methyl esters uu~ GLe

TLC

Methyl esters uu. GLC

HYdrogenated methyl esters uu" GLC

SterylEsters

Glycerylethers

Triglyceride Freefattyacids

Free sterol

___ ~ GLC Hydrogenatedmethyl esters

r

Saponify

Fatty acids

Methyl ~ GLCesters

Hydrogenatedmethyl esters

JGLC

Saponify

Fatty acids

Methylesters

-IHydrogenated

me:tl esters

Transesterify

Methyl • GLCesters

GLC

Methyl • GLC

~

Hydrogenatedmethyl esters

1GLC

Figure 1. Schematic representation of experimental procedure for lipid analyses.

OYSTE R (c. virginico)

T

PART OFTOTAL ESTERS 18:2""6

18: 0+

18: I

r--'---o",,9

""7

MOST MOBILEBANOS

80S

ADIENEBAND

LLJf/lZoQ.f/lLLJ0:

0:LLJC0:oULLJ0:

18:0\

18:0

""9",,7

TIME ----t

Figure 2. Comparison of totol C18 esters of C. virginico lipids (left) with nominal monoene (plus saNrated)ester bond (center) and a diene bond (right). 80S column, slightly diff.rent temperature on right-handgroup af peaks.

TTOTALESTERS

w(f)

ZoQ.(f)

Wa:a:woa:ouwa:

20:lw7

(20: 2)

20:0 +20: I

w7

OYSTE R( C. virginico )

BDS~

( 20:2 = NMI DIENES I

(20: 2)5,13 ?

TIME --.

Figure 3. Comparison of total C20 .sters of C. virginico lipids (left) with 0 monoene bond (center) and thediene bond (right). 80S open-tubular column, slightly c1ifferent temperature on right-hand group of peaks.

(7,17? )I .

(7,15 jl)"

(22:2=NMI DIENES)

OYSTER(C. virginico)

22:1",7

{"'13 11\22:1 ,./ir .

I 22:1(0)9

I

+- ex I 4X +

80S leOleo

~

20:5",3

PART OFTOTAL LIPIDSw

lI)

zoQ.lI)

Wa::a::woa::ouwa::

TIME~

Figure 4. GLC chart to show portion of non-methylene-interrupted C22 diene! relative to normal dienes. Normal22: 1 w13 + 22: 1 w 11 and 22: 1II) 9 were isolated from this oyster lipid, bYt shoulder thought to be 22: 1w7

was not OccOUflted for.

" :::::::::::::::::;t~<l~ClCl~ ;;.;;;;;;;;:;f I[nt:tr£tn~ tu:ut~n~. ~

oeeee!!ll::::;:;: ;,::S:::;:,;t 1~2 l:ol$:'t

Cl~;tll:;;;;;;

;;~ i:!:~~:i:;: ~~:~:::n= ,;:

,. oj

.. " ~ .. _e ~ ~ .... ~ .......

;;.,t ;;s ~ ~~~ r~;!~,

, :' e ........ e:-i: .. :-~ .. :- ...;:;;: ~ ;;:;~~;;:::::O:l::::~:::~

~ .. eeeee .. e:-::!::i~:Oi;:;::i:l

. e~eeeeee2' :=:::2'U:lIl: :::::! e;::~

i:!:'

,ee,;·u a ~~'

...... ,.~:!:e r e:

e~ .. , ......:;::::::; ;:;

........... e ... :: .. ~ .. ~:-~:;;;Ii:.l;:t,.;:;:; ';:~:;:::::8;:

e., .... _... t: .. _ .... __ •~;;.':;:;::!:!:;:;::~s::::se.::::

....... _e_ ..;:;I:!::~ee;

e ... e ... _ .. e:;:;;;::a:::: :!:::l::Cli:!

_eo e_ ... ll:~'=~ ;;;:;::.-:1: ~~:il:il <!l

._e.ee .. __ :::

;;:;:::;t,i~':"'::

.:-_::.o:- .. :-?:;:lj'~; :::::0:: :::=::~;;

....... _ -';1-

:~:r:8 :'!

"~~~?~?~ . - .. e_"=;:;;:;o~;~: , t:::::l: :;lie ~:O:~i:il~ , , • ~ r, ::: :; ~ "':::;:: ~::::

; -,; ,;; :';::l ::~ ~8::; ~2': ~ ~ ~ ~ ~~,;z =-;;::::(::!: :::::;:::l'l

, ,

, .,

o :.:

" .

Table V-B. UnU'IU" I non-.cthylenc-Intcrrupted fatty aclds hoillted frOll

O. HuH. total lipid and vulous lipid fnctlons be foro and after feedln. on D. tertl('llect•.

Qrtrea edulla, Control Ind Exper!laental fed Dunallella

Total Lipid Polar Lipid Triglyceride

Control Bxperluntal Control Expert.ental Control Expert.ental

(20:211)

(22:2s)

0.38

1.29

1.11

0.70

0.54

3.70

0.21

1.94

0.19

0.54

0.42

1.26

Steryl Ellters Glyceryl Ether Free Fatty Acid

Control EllperillCntal Control Expert.ental Control Experhaental

(20:2,)

(22:25) 3.02 1.81 0.72

"' ..... 1 ...1... 1 ,.",.•"J, hal..... ,- ~"qJn."'. '0'01 I,p'" ,"j,·."o., , .,',~ -, .... ~"q""'-'. lUo,.ol .noIl.,.......... 1 r.4 _1J.lJo

e., " -,q-.,--~I~ Coo'n>1 up<'r, ......1 CoB'",1 Eqert ....'.1 Co",,,,1 £oport_IOI c.o'n>1 !:.>;pe.._.al 0>0..<11 LLpo""UI

", '" .n •. Ol l.n .." ,.~ 1,11 l." '.n l.~l l."•.•• 1. ~..,., '" D.n ,~ O.ll ,~ O.ll D.ll 0.'1 "."l •. U... '" D.S' D,SI O.SS ,~ on ,.~ O.SI .,. '.'6O.'l

U...... " ..·130 '" 0.1l " . 0.11 D.H 011 0.11 0,07I."'" '" 1., ». 1.0: l,U '.1' I.Dl '" 1.1' ". L. 1.U l.n1... ·1.. 0 oc O.ll ." , .. O.l' 01O 0,11 OM 0.0l

U0.': O.l·1 •••IO.I"~ ,~ 011 ... 0."

1I.1l lS.ll :I.,," 1:.l1'.n

~.'n

n.n XI.'I", IS.O: .,.'I.ll ".n n.n1... ·170 1.11 ,~ ". 1.t>S ,. 0.1S I.n 1.15 1.U I.'l I.SI l.,'..1.....·110 ,. I.n D." 0.• ' ,.~ O.ll ,.~ .. ... 'N,', ,..., 151 .n l.l" '.Ol 1 .. 'N L' L. U

,n1 .•1 ,. ,..'.'.II.IS·"" , .. ",' 0.1. U I.ll O.IS I.ll1"..·1.... 0." 1.1' 0.10 0.1' I.'l ,n O.ll

'" .. '.fl I." ,. U '.1: '.16 ,. ,.. ".1'I' .• ." 0.1l ,n ,.. D." 0.11 U 0.0';0.1 ." .. '" II' 0."1:050....." 11.DI ., .. <>1,0"

lSI' 0.1. 1.10 ,11 ou1.1... 1.11 1.1. ,. ", 0.'1 ,n 1,0: 0.".l.·O

1.1.' .. 6.1' •. r ..•;I.S.: :..., ll,n

l,S'

".1"·0." ,~ ,~ O.CI 0.11

lilA 1111 " l.': '" .n .. '.11I.n

..L.

1.tI11,1.' ,n 10; ,.. ,. l.U 1,7l 1.1' I.~1I.I.l ,,' n n 0.:1 0.•0 '.U ... ,~ ,n 1.21 ,.O.ll

1"1' on U ,.O.I~

:0'1.11 ,n ,~ on ,. D.ll D,57 .. ,~ l.0l L. ,~ 0.11n ,~ • n " . L. D.'I 0.10 ,. 1.11 l.'l I.n l.U 0.09 'n1O:1..' .. ') S.O' ,n ,. ,~ U •. 11 I.n '.n l.H}O'IA '.M ." Ln I,n 0.03 0.11 '.S!in:f 1m '" O,IS ,.:i.:0.1. ,n ..

D." '",.

0.:',~ o.n 0,11

:.Sl '.~ o.nI," ... • n 'n O.OS ,. 'Nn .., 1'1lI IfU " .•S I'." 11,00 n.1l I'.n ".'1 11 .•1 ll.~ lO.to '.n1.0-3 on ,. ,. ". 0.11 ,.161'" '.11 ,n .n O.l: 0,11 ,. O.ll '.lS '.n,,~ , .. .., h

o.n0.11 0.61 ,. ,. o,n ,n O.ll 0." o.n D,1O1.:JwJ D.P 'n ,.

D.IS D,n ,. n O.ll 0.'11"_1 0.'1> •. n O.ll11,:..0 ,~ 1.11 '.n 1.11 OM l..S 1.1.11:1'" •. n O.OS 0.13 '.N1I,J..6 .~ ,~ ." '.M 0.10 0.1l 0.11I.:lo.l .n o.n ." LN 1.6S '.N I.Jl ,.~ I.ll I.U L~II • .,J lOS OM l.ll '.Jl '.N '.N 'N 0.'1 I.l'11,,.._. III I .•) ." 1,1S 1.01 LM 1.•0 O.lS o.n : ..:E:EI'3¥9J , .. 'N ...0.01

:.ll 0.15 D,IS ... 0.1' 001 '.n 01' '.M 0.'10.0: ,. , .. o.os lI.n '.n 'M 0,07 '.Il 0.11O.IS

D.H 0.0' O.ll '.n 0.11 D,IS 0.1. 0.11 0."1O:'.,)

1.23 • n ...'N 0.0l

.. I.n 'N 1.'1 D.1I l." ,~ l"1 1.SS.. .., 0.11 D.'S O.IS 0.11 ... 0.17 11.1' O.lS ..~lO,So.) .., ", ,. 1.1l ..0.2'

10.56 12.ID •. 31 0.10 ..16 'M 11.06 10.5.010,,"'ho,,", 0.01 1.11> I." l.~: O.ll 0." 0." 1.61 2.81 O.S' .."11:" ,. ~. II 0.2S 0.120.11

0.57 0.61 0.10 0,'1 D.lIl ..~ 0.:1>12:..... ,. 0.02 0.2S 0.11 •.n ,.. 0.01o.nn:l.6 0.15

'N..,

0.10'.m .~ o.n 0... 0.1'

0.l70." D.ll11,SooJ D.ll ." D.ll 0.1' 0 .•1 o.n 0.)9 '.n 0.1.

•. U11:1>003 S.O) D.n ~1! ,. .. IS.IS 1l.71 l .•' l.a

1.37 '.N '.nn'""b.-.. I,n ". '.n o. 1~ 0.2. 0.17 ... '.Nll'ollcno 10.15 ." 20,'5 26.0\ 16,11 SO.19 ... 01 25 " Ill.$:! lO.pS JJ.P' 31." 3325

f'"l,,,h"d " I"~ ". ..

frnllo.... l.~ 'M ."hou' r.... ,•• ""D. ,.,tJ.j_~ • VI<! .r... oh ...,~. 010<>1'.

',·.....«_.;...,10,<•. "'.'rol u6f..""I ...uI '" .......11.11.

T·'rl1<.r,~. SU.,I h .... (••h~..hl

~ ... , .... H ... ['I'. H .~. ,"",..,01 I...,..l_ul .....IE.<porl ...UI C••" ... I [''1''''''''101 ",*, ...1 Exp<t.'......1 """ ...1 Eo:porl ......1 """"'1 Eo:po.I_..l e-'.... l Er,>r""o'OI

~,U 3.Z' 'M .~ ." •. » •.•1 •. Il ..• 1.•1,Io.n •. S3 10.IS0.1. o.r. 0,70 0 .•• .~ 0.•' .~ o.n 0,'1 o.n o.s' .u Z.•'D,lt O.ll 0.•' '.M 0.'. 0.'1 D.•' W .~ 0,51 0» 0.11 •.u L"O.lf ..~ .~ .., .» ." o.u O.U 0.1' 0./1 .~ O.'S l.ll '.MI.U 1.17 ,.~ 2.10 I.n L~ I. ~S 1.63 I.ll LM ,~ Z.ll ,.~ ,.~O.ll O.lZ 0.'1 0,31 .., U O.U U 0» 0.1' ." 0 .• ' O.lS a.•'n.n n.u 51,'1 ,...•. ". 0,.0

u .• 3S.~ ~. 2..... 1'.39 n,.3 Z'.·S.." 1.13 0.'3 L~ .,.. n.t!>I.U 1.51 ,." •. n 0.11 0.S30.•1 0 .•1 0,.7 ... I.Jl •. » •. n •. n O.U O.S' o.n ... 'M.." ,.

'M ,~ 2.'J /'.1 1.11 2.05 I.U lo.ll 'M ,.OOJ • 11 •. Z• ... 0,51 O.Sf 0,1'0.15 ." 0.13

'·11 .. 1·'5 '.51 .... 0.51 •. n '.01 0» '.1'·.11 l." '.15 ,. ,. 1.Z' '.u ,." 1.51 I'.n .." 5.11." 0.10 '.M 0.11 .." ..~ 0.'7 .".~

n.OJ 0972 Sl·72 Rl. n.so '0."0» 0.11

L~..~ '.M .~

JSl '.M ..... 1,15 0.93 1.2' 1.J1 L. .. I.ll ,~'.n 1.13 lo." 10..10 '.n lus ,.. '.'0 ..11 llOU ;1.... ill... lI.n..~ ." 0.61 •. n 0." ... O.Ul,21 ,u I.n .. ... ,. l.U ..n '.11 u 7.'1 I •.•• 11.79,.~ I.U ,. '.n 'M 3.S1 '.M '.21 1.72 I." 1.J1 L..U 0.'1 0,S1 0» '.3' O.lJ 0.'0 .. O.lf ..n on 0.15 0.11 •. n'.10 o.M O.IS 0.1S .... ..,.., .. • n ,. 'M L. ... 0.11 1.1. ... ,.» I.IJ•. Tt on .. '.Sl •. 02 ,.~ •. n '.1' 1.01 L~ ... ... 0.12 1.21,.~ OM 'u ,." L. Z." 1.13 LM •. » 1.01 I.U '.M

0,11 0» '.2' •.» 1.15 '.M0,'0 •. 21 ..• O.la O.IS ...0.03

OU ..~ U L~ 1.17 •. 1' 0.1I '.21... ." .. •.U ..• 0.11 ..~ ..~ .."1'.lI 1',N n.n n.u n.l. 21.lS /1." 71.11 '.M ZI.OI 03.... .uo.n 0,31 ... 0."0.1' 0» ...

..u •.uh '.M

0.'10.21 .." 0.11

0.13 o.n0.1' 0.01 0,11 O.U ..~ O.SJ 0.15 0.31 O.U 0.33O.U 0.31 .." 0.11 '.M 0.1' ... .." .~ 0.11 0.51 U0» O.JJ

2.U 1." 1.Sl 1.•' '.M L. I.UD.51 1.510.10 0.17 ... 0.10 .." 0.01 l·.n' ......• .. 0.11 O.ZS .." 0.11 D.n 0.1l1.12 ,.» 0.31 1.21 1.'1 1.11 L~ 2.Sf 1.Sl ~.Jl o,n ..~ 1.1.I.U 0 ... I.•• '.n 1.01 1.01 3." 1.11 2.n '.M 0.61,." 5." 1.73 l.n L. ,.~ l.n '.u 1.12 ,."0.2& o.n 'M 0.11 O.lS 0.12 0.1' U 0.31 o.lT0.05 O.U 0.10 0.03 ..• .." 0.11 0.05.~ .." 0.10 0.01 .." .» 0.03 0.01 0.13 ... 0.1' ..•

0.11J.U 1.1l ." O,~' 0,71 •. u .." 1.1l 0.13 0.51 O.OJO.Sl .." O.OJ ..• O.SI O.U 0.10 0.3J 0.01 .. " ... 0» ...11." n.IJ u D.'l ... •• 11 •• 13 '.03 '.71 6.111 L"L. 0.65 •. 61 0.16 o.n 0,'0 O.lI0.01 0.25 O.OJ O. l~ 0.11 .." 0.19 o,n 0.110.01 •. n 0.11 D.Ol 0.01." O.U .." 0.2~ ..• 0.0' G.n 0,1' 0.01 0,"0... 0.16 ..~ o.n 0.1' 0.1I 0.05 0.31 0.1' 0.10 0••0 o.un .., n.1l 5.51 J.16 ,.w ..~ '.31 '.61 0.51 L~0.11 2.12 ..~ 0.21 O.OJ"60 .5.12 10.52 20.'5 ".111 n,60

r.~lo VI·A. _ry" dolol. I...."" ..' (oUr ...1.. , ... lo.oM ,..- 1:. nr,rJ•.u••oul lipid ..a ..._~ ..U'I..... eo.t....l _ Upcr'-a'o' ,. -.LIol~

COn .....1 Exporl_nUI ;::~:::••:o,..",_=...:::.::;u, u-.-,.-,-..-.,-,.-..-., hI'. I £>:1'.1

Table VI-B. Unusual non-llethyll!ne-interrupted fatty acids isolated frOli C. vlrglnlca

total lipid and various lipid fractions with and without feeding on

D. tertJolecta and after six .onths stonae.

Cra51f~tr.. vlrglnlca, Control and Experi_ental Fed Dunal1ella

Exp. t

Initial

Exp. II Exp. I

6th .onth

Exp. It

Control Experi.ental Control Experi_ental Control Experl-ental Control Experi_ental

Total Lipid

(20:2s) 1.00 0.82 1.01 0.98 0.54 1.18 2.09 1.49

(22:2s) 4.57 2.11 0.87 2.55 0.73 0.58 1.82 1.55

Polar Lipid

(20:2s) 0.86 0.92 0.15 0.63 0.45 2.19 0.86

(22:2s) 6.32 2.90 6.70 5.49 1.45 1.13 2.88 1.49

Trialycerlde

(20:2s) 0.61 2.12 1. 79 1.59 2.16 2.11 2.40 2.43

(22:2s) 0.28 1.71 1.11 0.78 0.61 0.89 0.90

Steryl Esters

(20:25) 0.71 1.02

(22:25)

blllrVll. "-or.11 lin I... 1 f"lly"c:ld, I ~n'''trd rroo D. ftd"l1_ tOUI HI'

llltnM ,'(/"/f,,, C"lItr,,1 "nd "_I'rd"'1l1,,1 1.,,1 '."Chl"t/.lllnutl llIc .... t,.rllI

TOI:tl 1,ll,ltt l'e,I" .. 1.I['id

bl'f'ri ..nlltl "al><' r ~..It t n I hrcrl ..nt,,1 1_I>rr.-rnt:tl

Control r", lllochr'ldll rrd Dlcrd.rJ. rt'd Uochr\lsb fC'd Dlcuted.

I~ ,0 1.67 15.24 10.90 S.1l 1.02 7.01 I.OS4.11.12·nrrn 0.11 1..10 0.97 0.9S 0.70 0.47 0.69

". 1"'0 0.11 I. ... 1.47 0.114 0.92 0." 0.17"nld~" -15:0 O.M 0.82 0.52 1.44 0.71 0." 0.1315,/1 5.59 fl.S6 (;.71 \.\7 '.06 3.'] 2.06I-·nll,-O o. 0.57 n.65 1./.7 1.58 I.,. ....J.I',III.I .. m'nIf,,O 'I .... \9.31 31.75 ](1.01 25.12 27.19 01:.221...'-17,0 2." 1.69 l.OS 1.42 1.16 1.56 I.fl9Anl,·j ~.. 17:0 0.64 1.9] 0.71 1.111 4.61 0.71 0.6511:0 4.61 \.62 5.17 2." 2.19 2.03 \.Ott\.7,ll.15-TNllt' ..."IIt:O 1.10 0.10 2." 0,47 0.40 0.46 0.29111:/1 19."1 10.67 20.17 II.lIf, 12.24 12.'5 7.4(1I~"l 0.011 0.10 n.1I 0.01 n.1210'0 0.49 0.16 0.21 0.60 0.11IJ,II 0.0\

".'Inr:tlrd

1.. ,11.. ,1..9 L5 0." 1.92 1.42 4.39 1.70110'1 ..7 :.\." 4.02

l3.:\7 :2.\6 :2.31 : •. 695.95

Ih:I,,5 0 .... 0.1711:1 •.117111;1 .." ""11) 1.11 1.111 1.4~ 2.49 ..... 5.61 5.411,tI,1 ..1 I." 1.14 1.10 1.21 0.79 1.23 5.91111:1",5 Ira!;: 0.06 0.04 0.20 0.15 0.291!1:19 0.10 0.4\ trllce n.'" 0.15~O; 1.. 11 0.47 0.]5 0.27 0.'" 1.22 0.83 0.51~n: 1..9 0.17 0.-4\ 0.\1 0.72 0 .... 0.76 O.H'n.I ..7 1.73 0.18 n.1I> 1.(17 I.foil 2.01 2.511211:1 ...... 0.16 0.10 O.()(, 0.\0 0.1. 0.23 0.4012,1 .. 11 r·..I\l O.l' O.M 0.02n;I.~) 0.19 II. nil O.U 0.11 0.1]lZ:I",1 0 .• ' I. Ito l.2S 1.95 0.62

12.(,5

1/,:1,,", 0 ..\2 0.46If,:.\..1 O.IUl 1l.19110:]..,11/,:4.• 1 0 . .(7

1ft: ~..", O.H 0.011 0.26 0,82 2." 3.91 1.\1IM:J.'" 0.29 0.79 n.4~ O.(,~ 0.01 11.0\IH' "'~, 11.29 U.118 n..!~ n.os O.OK 0.""III: ,,~, l.h.l 0.81\ l.4b ~ •(of, >\.20 2.83 l.IOIH:>\.d 0.92 O. 2~1 0.\.' I.HII 1.03 0.85 0.61

2U,1.", (. _~.9 I.llt, 1.32 0.01

211: "'~, O.W 0.0\ 0.01

/!I''-' 0.411 n.19 0.:\1 0.24 0.30 0.01~II:"-"h 0.11 O.1l7 O.IIS ~.bll 1.93 1.41 0.45'Ihl,,':\ ."" 0.11 O.lIl 0.12 O.OS 0.11

':11:5...:\ "4' n.lO 10.21 6.1>2 b.45 2.16

21 ~. ! 0.02l.l',I,.(, 0.0921;5.·,(, 0.27':2: ~....1 o.n 0.l4 0.11 0.1l 0.02n,r_' O_Sf. '.2') 1.7<1 3.78 O. 9~

1\,1 ~'-lU' l2.:\2 21.21

l·ol,·"I'''-,1 l.V, n."s

:and ..... riOU5 Ilr ld rnetluM with lind without reN.nllln '" ....lb"'Mllnd .. JnorIYU.

Oller" "",,'f., co." 1 • ,tdflpt'rl.enUI Ff-d r.odlrv"' .• .ndDt"r.tllrJ.

Tr1Illyl'C'rldf- Fn'C' Flllty Acld~ Stl',-"I F..HI'r5

I'~perl-ent.1 I'xl'~rl.entlll IXI'l'rl-entnl hperi.C'nllol hllllrl_nul l:xpl'rl-ent:>1

r,d r"ochrllsJ. fed Dlcnt.rI. Control 'od t.ochr~"fll fed Dlcrater'. fed rllochrllsh fedDJcrllterlll

5.1S '.00 6.91 1.35 4.42 12.13 2.42 11.360.4' 0.12 1.56 0.08 0.65 1.\9 0.68 1.490.69 0.94 0.79 0.12 0.61 1.87 0." 1.590.13 0.11 0.25 0.05 '.30 0.72 0.21 0.571.71 1.88 12.81 0.69 2.96 5.16 1.97 2.730.42 0.67 1.12 0.48 1.14 2.97 0.85 1.35

18.01 40.49 15.12 31.04 37.12 35.86 31.69 30.941.111 1.43 1.55 1.78 1.78 1.41 2.115 1.79O.M 0.51 0.53 1.07 0.54 1.00 2.18 1.5t.2.11 2.56 4.78 '.40 11.93 1.06 l.85 0.7'

0.25 0.25 0.96 0.24 0.43 0.'1 0.50 0.331.15 1.59 1.'0 21.26 11.'1 5.73 6.13 s...0.01 0.01 0.14ll.l\ 0.19 O.B 1.79 0.99 0.68

0.491

M.1I4 15.21 68.35 15.11 10.6\ 52.95

1.11 1.84 1.16 0.63 1.01 5.11 5.03 3.806.22 6.14 3.81

:0.952.68 5.11

:7.959.38

0.50 0.61 0.19 0.11 0.47 0.45.... 4.11 5.21 4.89 3.19 6.91 9.81 6.196.42 7.37 3.69 3.26 3.68 2.19 9.05 6.350.16 0.20 0.190.23 0.040.50 0.47 0.10 1.54 I.M 0.74 0.90 \.030.26 0.40 1.05 2.61 1.04 0.63 0.90 1.14::.73 2.31 1.28 8.46 3.64 1.25 3." 0.980.29 0.25 0.23 0.21 0.07 0.49 0.21

0.11 0.380.09 0.12 0.05 0.11 0.03 2.79

0.42 0." 0.42 0.38 0.41

23.86 2<>.04 22.81 17.10 22.80

0.35 2.13 0.53 0.16 0.64 1.05 0.170.25 0.30 0.24 0.47 0.43 0.26 1.080.03

1.85 l.25 0.31 0.08 0.52 2.110.12 0.07 0.02 0.010.06 0.04 0.031.84 1.08 2.60 4.62 0.77 1.01 0.84

1.26 0.51 0.07 0.04 0.15 0.39 0.2<>

0.05 0.010.04 0.06

1.11 1.20 1.630.07 0.02 0.120.76 0.54 0.24 0.35 0.64 0.73 0.24

0.31 0.072.216.16 2.2<> 0.11 0.13

0.092." 0.18

16.08 9.04 8.82 6.58 5.62

48.29 30.06 41.90 31.81 59.10

ndvari(ll,l5 lipid fraction,wnh.nd ..ithout feflhnlonr'9'1~andZl.J""'T1IIIU.

o.t ... , ""..Jh. e.:.ntrol and hperl.-nul Fed rloehryah and DJcTlt.r/,

Free fatty Acid5 Steryl Euen

5.11 1.00

1.71 I.U

S1.SJ

23.7. 22.93

1.61

6.97 1.3S 4.U 12.83 2.42 11.:56

12.81 , 0.69 ,." 5.16 1.117 2.71

33.26 42.64 1S.1l3 44.79

4.71 0.7'

37.23 26.15 16."

6.44 7.53 6.52

0 . .(9 0.93 3.16

Table.\'IJ·A. ~ry of chain lenJths of fatty atid, holaad f". o. -.;full_ total

_1.0-.. eduJia. Control and ~ri_nUI 'od Z_odIryai_ and Dierauria

Toul Upid Polar Lipid

Exper1_nul Experillenul Uperiwnul Experillental

fed laochry.Ja fed DJeTatada Control red laoc:hryai. fed DJer.tad. COntrol

'I< 8.67 15.24 10.90 5.83 7.02 7.08 8.05

'IS S.Sg '.SO 6.71 3.37 3.0. 3.83 2.06

'" 37.64 46.28 34.27 32." 33.0!l 50.67

'" 3.62 5.77 2.36 2.8g 2.03 3.06

'" 26.80 2S.S6 22.73 26.03 27.S6 22.22

'" 0.10 0.51 0.10 0.11 0.52 0.15 0.12

'"7.67 3.gl 3.'" 17.37 13.34

'" 0.02

'"l.48 0.02 0.22 '.00 3.60 S.g8 1.71

"''1. VIII No_I ••1_' ,.", .<ld. 1001> f t'. .'r"al.......1 IIF'd. """ ••d .... lIF'd fTO"I .... vlt~ 0lI<l vl._. food'"1 .. I _ID.I_••

C• ..-u_ ....,.._. e-.....I -.I lqa,l_ul ,0<1 ,_....,. _III~ ,.. c.._u.. 0'''''0''', e-...., oaoI ~.,.'I .....l fod I~.'......111 '.

~d_••1 l.,.rl_..1 bpnl_UI 1""'MOtol --==::-:--==::;_ ..... , fod '-.,.,10 fod 01.......". _.",1 fod '_I. fod ~.......,I. _.",1

',05',U

I.U__

,d_"".,I',,'·~

..10....11,.

1.1I.IS·'"

• u,...

,.....•.U

,.'.S1

,.1.1•,.

...)

..'.tI

'.1',.'.tI'.1'

...'.1.

...)

..,..."U.tS

I.U•. U

'.ff...)

....,.•. ll

,..''."•. 01'.OJI .•'

• .•1

o.n

,~...,

_"'0' thai" I.III.h. 01 ,.ny acid. 1101..... ,.,.. C. ,bll'J~I"," ....1 lipid........1..... lIpl. , ....1""••I,h wI,,,,,,,. 'oodl"l 00 I. ",'kno , D. 1_......

e............ ''',''''",", <'- ....1 ..... txpo,l ...tal F" 1__10 """ Nera.ul. cr._or.. ""Imea COO I oM to...I...... ' , .. ro_rya.lo &IWI1tl«ratul.

to l...to' (.,.,I tal (","1'1"'''1 (>po.I 1 tqo.l.....1 tJlparl ...tol f_,I .....1 txporl 1

COol.... I f _I. f .. Dle I. Cooo....1 F'" ,_J. '... DI"' I. COol .... ' F... ,_b '''0''''''.''. C>a ....1 F... ,_Ja F....J ··,,·

'abl. VIII-I. UnU~UIlI non-_thyIVle-inuTT\JPud fatty .dd~ holaUd he.

C. virgJnJI:. total lipid~ and vanous lipid frattiOll~ .,Ith and

.,i thout feedin. on 1. ~lhanlo and D. inorlWlt..

Cr...._er.. .,Jrgillic., COIltrol and ~eriM1lUI fed l~brll.j. and Di~At.,.j.

ExperiM1lUl ExperiM1lul

Control Fed J.oebry.i. Feel DJer.tu-J.

Total Lipid

(20:2~) 2.31 I." 2.02

(22;2~) 2 .•' •• OJ 2.13

Polar Lipid

(20;2~) 1.19 1.27 1..6

(22:2~) '.09 6.39 6.63

Tri.lytulde

(20;2~) 2 .• 2 1.87

(22: 2~) 0." 0.67 0."

Suryl Ener~

(20:25) 2.53 en(22:2~)