34
Aquaculture
ELSEV IER
A quaculture 155 (1997) 1 4 9 -1 6 4
Review on the dietary effects o f phospholipids in
fish and crustacean larviculture
P. Coutteau a *, I. G eurden \ M .R. C am ara a l, P. Bergot b,
P. Sorgeloos 11
J L aboratory o f A quaculture and A rtem ia R eferen ce C enter, R o zier 44, B -9000 Gent. Belgium
b U nité-M ixte IN R A -IF R E M E R de N utrition de Poissons. Station d'H yd ro b io lo g ie. BP J. F -6 4 3 W Saint
P ée-sur-N icelle, F rance
Abstract
A beneficial effect of dietary phospholipid (PL) supplementation in purified diets in terms of
survival, growth, resistance to stress tests, and occurrence of deformities has been demonstrated in
larval and juvenile stages of various species of fish and crustaceans. The exact, determination of
PL requirements in larvae is complicated due to the difficulty to bio-encapsulate PL in live prey.
Furthermore, the great variety in purity and composition of the PL sources, and the experimental
conditions (such as diet formulation and extent of co/prefeeding with live food) makes it difficult
to compare requirements determined with artificial diets. Larval stages are extremely sensitive to a
dietary PL deficiency and require higher levels of dietary PL than juveniles. For most of the fish
and crustacean species examined, the estimated PL requirement of larvae are in the range of 1-3%
phosphatidylcholine + phosphatidylinositol (PC + PI) of diet dry weight. The absence of a PL
A bbreviations: Choline CD P: C holine c y tid in e o '-d ip h o s p h a te ; D H A : D ocosahexaenoic acid (2 2 :6 « —3);
EFA : E ssential fatty acids; EL: H en-egg lecithin; EPA : E icosap en taen o ic acid ( 2 0 :5 « —3); EPC: H en-egg
p h o sphatidylcholine; FS: Free sterol; H UFA : H ighly unsaturated fatty acids; NL: N eutral lipid; LPC: 1-Acyl
lyso phosphatidylcholine; PC: Phosphatidylcholine; PE: P hosphatid y leth an o lam in e: PI: P hosphatidylinositol:
PL: P hospholipid; PO L: P olar lipid; PS: P hosphatidylserine; P U F A : Polyunsaturated fatty acids; SE: Sterol
esters; SL: S oybean lecithin; SM : S phingom yelin; S PC : S oybean phosphatidylcholine; SPE: Soybean phosphatidylethanolam ine; SPÍ: S oybean phosphatidylinositol; L ecithin: A lthough the term 'le c ith in ’ is the trivial
n am e for ‘phosphatidylcholine’, lecithin is defined in the food secto r as a mixture o f p olar and neutral lipids
w ith a p o la r lipid content o f at least 60% (H ertram pf, 1992). In the p resent m anuscript, the latter definition o f
‘le c ith in ’ (e.g., EL, SL ) was follow ed, w hereas ’ p h o sp h atid y lch o lin e’ is used for the specific PL having
ch o lin e as the headgroup (e.g., SPC, EPC).
C o rresponding author. Present address: IN V E T e chnologies, O ev erstraat 7, B -9200 B aasrode, B elgium .
T el.: 32-52-331320; fax: 32-52-334531; e-m ail: p.coutteau@ inve.b e
1 P resen t address: E m presa de P esquisa A gropecuaria d o R io G ran d e do N orte (Em parn), C aixa Postal 188,
5 9 0 0 0 N atal, RN , Brazil.
0 0 4 4 - 8 4 8 6 /9 7 /$ 1 7 .0 0 © 1997 E lsevier S cience B.V . A ll rights reserved.
PH S 0 0 4 4 - 8 4 8 6 ( 9 7 ) 0 0 1 2 5 - 7
Reprinted w ith permission from: Elsevier Science
35
150
P. C outteau et a l./A q u a c u ltu r e 1 5 5 (1 9 9 7 ) 1 4 9 -1 6 4
requirement in the freshwater prawn Macrobrachium rosenbergii exemplifies the important
species differences. The few studies evaluating single PL demonstrate that PC and PI are the most
efficient in most species. The presence of an unsaturated fatty acid in sn — 2 position of the PL
molecule seems to be essential for the functionality of PL. Some studies in crustaceans reported a
relation between PL requirements and the protein source in the diet. Various hypotheses have been
formulated to explain the effect of PL. The PL effect is not related to the provision of choline,
inositol or essential fatty acids (EFA). However, PL may be superior to neutral lipids for larvae as
a source of EFA and energy due to their better digestibility. PL may improve the performance of
the diet by improving the water stability of food panicles, or by their action as antioxidant or feed
attractant. The effect of dietary PL appears not to be explained by their emulsifying ability.
However, there are proofs that dietary PL interfere with lipid transport, especially cholesterol
transport in crustaceans, and with retention of fatty acids provided by dietary triacylglycerol.
Although the origin of the requirement is still unclear, dietary PL supplementation has potential
importance for the formulation of practical larval diets. © 1997 Elsevier Science B.V.
K eyw ords: P hospholipid; Fish; C rustaceans; Larva: Juvenile
1. I n tr o d u c tio n
B en eficial effects on culture perform ance due to the inclusion o f phospholipids in the
d ie t w e re first reported in the late seventies fo r lobster (C onklin et al., 1977) and larval
fish a n d shrim p (K anazaw a et al., 1979, 1981). T hese findings w ere surprising, as de
novo synthesis o f phospholipids has been show n in various species o f fish, w here it is
g en erally b e lie v ed to follow the m am m alian pathw ays o f e stérification (H enderson and
T ocher, 1987; Sargent et al., 1993), as well as in crustaceans (Shieh, 1969; Chapelle et
al., 1985; T esh im a et al., 1986c; K anazaw a and K oshio, 1994). D espite substantial
research efforts encouraged by this apparent discrepancy, the m echanism s behind the PL
re q u ire m e n t are still not fully understood. T his p ap er review s the present experim ental
e vidence o f PL requirem ents and the various hypotheses fo rm u la ted concerning their
role in larv al and early juv en ile nutrition.
2. P h o s p h o lip id re q u ire m e n ts
2.1. P a rtic u la ritie s a n d constraints in studies on P L requirem ents in larvae
D ue to the lim ited acceptance o f artificial diets at start-feeding in m any species of
larval fish and shrim p, m ost studies determ ining larval nutritional requirem ents for
essential fa tty acids and vitam ins have been carried out using en rich m en t techniques to
m an ip u late the com position of live feeds such as rotifers ( B rachionus p lic a tilis:
R ainuzzo e t ah, 1994a) and brine shrim p ( A rtem ia sp.: L ég er e t al., 1986; Sorgeloos et
al., 1997). Initial studies on larval phospholipid requirem ents attem pting to bio-encapsulate p h o sp h o lip id s in A rtem ia failed due to the general conservative nature o f polar lipid
class c o m p o sitio n in live organism s (T ackaert et al„ 1991). T he latter was confirm ed
recen tly b y R ainuzzo et al. (1994b) w ho found only m in o r shifts in the lipid class
36
P. C outteau et al. / A quaculture 155 (1997) ¡ 4 9 - 1 6 4
151
c o m position o f A rtem ia and rotifers enriched with an em ulsion based on e ith e r halibut
roe or ethyl esters (co n tain in g 71.2% and 0.8% o f PL, respectively). A lthough it m ay not
be possible to obtain live feed containing ranges o f phosp h o lip id levels suitable for
nutritional studies, fu rth er research is needed to evaluate the effect o f m in o r m odifica­
tions in the lipid class c o m position such as the PC :PE ratio in A rtem ia (R ainuzzo et al„
1994b) on its nutritional value.
T he exclusive use o f artificial diets from start-feeding onw ards is restricted to only
few species, e.g., the goldfish C arrassius auratus (S zlam inska et al., 1993) and com m on
carp C yprinus carpio (R adünz-N eto et al., 1994; G eurden et al., 1995c). w hich readily
accept sem ip u rified diets upon first opening o f the m outh. By contrast, m ost studies
have been carried out w ith larvae and ju v en iles initially fed on live feed, w hich is
relatively rich in PL (T esh im a et al., 1987). T he live feeding p eriod varies from a short
prefeeding on m icro-algae (e.g., feeding diatom s till zoea I / I I in larval Penaeus
japonicus', K anazaw a et al., 1985a) or rotifers (e.g., till 10 days a fte r hatching in larval
ayu P leco g lo ssu s altivelis', K anazaw a et al., 1981) to a com plete larval rearing on live
diets and con seq u en t w eaning onto the experim ental diets (E uropean sea bass D icentrar­
chus labrax: G eurden et al., 1995b; P enaeus sp.: C am ara. 1994; C outteau et al., 1996;
lobster H om arus am ericanus: C onklin et al., 1980).
A n o th er difficulty is the nonstandardized com position o f the e x perim ental PL sources
that differ in th eir c o n te n t o f neutral lipids (e.g., < 1% o f total lipid in de-oiled SL
versus 46.5% in raw SL ) as well as the proportion o f different PL c lasses (e.g., 23% PC
versus 42.1% PC in tw o types o f de-oiled SL) (data reported by C am ara (1994) and
K anazaw a (1993), respectively). Furtherm ore, variation o f the level o f dietary phospho­
lipids has been c o m p en sated using cellulose (e.g., K anazaw a et al„ 1983b; H ilton et al„
1984; T esh im a et al., 1986a) o r neutral lipids (K ean et al., 1985; C am ara, 1994; G eurden
et al., 1995c; C outteau et al., 1996). T he latter results in isolipidic diets and has been
preferred in m ost recent studies. In addition to being isolipidic, diets designed to
com pare the nutritional value o f different PL sources have been form ulated to contain
sim ilar levels o f p hospholipids (K anazaw a et al., 1981; G eurden et ai., 1995a,b,c),
PC + PI (K anazaw a, 1993) or PC (C am ara, 1994; C outteau et al., 1996).
2.2. Influence o f stage a n d species
T ab le 1 sum m arizes the PL levels reported to be optim al for several larval and
ju v en ile crustaceans and fish, and the relevant experim ental conditions. T h e criteria used
to ev alu ate the nutritional value o f PL consist m ainly o f survival and grow th, som etim es
com pleted w ith data on eith er skeletal anom alies, stress sensitivity, body com position or
lipid an aly ses o f the anim als. L arval stages are very sensitive to PL deficiency. Zoea
I / I I stages o f P. ja p o n ic u s suffered 100% m ortality before reaching the m ysis stage
w hen fed a P L -deficient diet (K anazaw a et al., 1985a; Fig. 1). F o r first-feeding carp
larvae, the su pplem entation o f 2% PL to the diet highly im proved both survival and final
w eight (by a fa cto r 2 .0 -2 .5 ) com pared to that o f carp fed PL -free diets (R adünz-N eto et
al., 1994; G eurden et al., 1995a,c). S im ilar drastic effects on survival w ere show n for
larvae o f g oldfish (S zlam inska et al., 1993) and red seabream (K a n azaw a et al., 1983a).
T he su rv iv al o f ju v en ile fish appears to be less affected by the d ietary PL supply than
37
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F ig. 1. G row th and survival of shrim p larvae (P . ja p o n ic u s ) fed a d ie t co n tain in g 0 c/c I O ) o r 3.5‘/r ( • ) o f
soy b ean lecithin (from K anazaw a et al., 1985a).
th a t o f larvae as show n in rockbream O plegnathus fa s c ia tu s (K anazaw a, 1993) and sea
bass (G eu rd en et al„ 1995b). B esides their effects on survival and grow th, dietary PL
w ere fo u n d to influence the occurrence o f skeletal an om alies and stress sensitivity.
K an azaw a et al. (1981, 1983b) indicated that larval ayu m ay require phospholipids in
th e ir d iet to prevent the incidence o f m alform ations, especially scoliosis and jaw
d efo rm ities. A lso, first-feeding carp fed eith er on a P L deficient or PC enriched diet
ex h ib ited a h igher incidence o f spinal deform ities than larvae fed a PI enriched diet
(G e u rd en e t al„ 1995a). T he vitality index o f larval rockbream fed 5% SL w as three
tim es h ig h er com pared to that o f the group fed the P L -free diets (K anazaw a. 1993).
P o stlarv al P. ja p o n ic u s fed a PC -supplem ented d ie t ex h ib ited a higher resistance to a
salin ity stress than anim als fed a PL -deficient d iet (C am ara, 1994). This PC effect on
stress sensitivity could not be confirm ed for P. v a n n a m ei, suggesting that this criterion
m ay n o t be suitable for evaluating the nutritional status o f euryhaline species (C outteau
e t al., 1996).
T h e PL requirem ent generally decreases w ith age o r developm ental stage, as show n
fo r ro ck b ream w here required levels decreased from 5% to 3% o f soybean lecithin
(e q u iv a le n t w ith 1.6% to 1.0% PC + PI) from larva to ju v e n ile (K anazaw a, 1993). T here
is so m e confusion over w hether excessive levels o f d ietary phospholipids can be
d etrim en tal to fish and crustaceans. Increasing the d ietary PL level beyond the required
level d id not affect survival or grow th in various studies (C onklin et al., 1980;
K an azaw a, 1993; G eurden et al., 1995c). H ow ever, survival o f larval P. ja p o n ic u s
d e cre a se d w hen m ore than 3% soybean lecithin w as added to the diet (T eshim a et al.,
1986e), and C outteau et al. (1996) (Fig. 2) fo u n d lo w e r grow th o f postlarval P.
v a n n a m e i fed 3% pure PC com pared to the shrim p fed 1.5%.
F ro m T able 1, it is clear that PL requirem ents m arkedly differ betw een species. For
instance, fo r ju v en iles o f different penaeid shrim p, the requirem ents are m ostly w ithin
the ra n g e o f 1.2—1.5% o f active PL (PI + PC ) (T ab le 1: C hen and Jenn, 1991; Chen,
1993; K anazaw a, 1993; C am ara, 1994; C outteau e t al., 1996). F or ju venile flounder
( P a ra lic h th ys oliva ceu s) and rockbream , these levels w ere 2.2% and 1% PC + PI,
re sp ec tiv e ly (K anazaw a, 1993). In contrast, fo r the fresh w a te r praw n ( M acrobrachium
40
P. C outteau et a i / A q uaculture 1 5 5 (1 9 9 7 ) 1 4 9 -1 6 4
155
rosenbergii), no significant effect w as found o f dietary lecithin supplem entation on
grow th n o r survival (H ilton et al., 1984; B riggs et al., 1988; K anazaw a, 1993).
2.3. In flu en ce o f the lipid class com position o f the P L source
A typical o il-free soybean lecithin contains 84% o f PL, including 22% PC, 23% PE,
20% PI and 2% PS (H ertram pf, 1992). N evertheless, a w ide variety o f PL sources has
been used in nutritional studies (see 2.1) and only few investigators have determ ined PL
requirem ents using highly purified PL sources, such as 80% (C hen and Jenn, 1991;
C hen, 1993) o r ev en 95% pure PC (T akeuchi et al., 1992; C am ara, 1994; C outteau et al.,
1996). T h e few studies evaluating purified PL fractions sug g est that PC and PI are the
m ain gro w th -p ro m o tin g fraction in lecithin. C om paring various PL fractions derived
from b o n ito egg, K anazaw a et al. (1985b) found that PC (purity: 42% o f total PL) or PC
plus PI (52% + 40% o f PL ) gave good grow th and survival rates in larval ayu, whereas
PE (6 3 % o f PL ) w as less effective. As 1% dietary supplem ents to pollack liver oil (7%
o f diet), soybean PI and soybean PC w ere m ore effective in im proving grow th and
survival o f larval P. ja p o n ic u s than bovine-brain PS, bonito-egg PE and bovine-brain
PE, w hereas a bovine-brain SM supplem ented diet did not perform any better than the
P L -d eficien t d iet (K anazaw a et al., 1985a). In a sim ilar study, soybean PE (95% PE of
total P L ) w as inferior to soybean PI (60% o f PL ) and PC (68% o f PL ) as far as survival
o f larval P. ja p o n ic u s w as concerned (T eshim a et al., 1986e). H ow ever, the relative
im portance o f PC and PI is not necessarily the sam e fo r differen t species. T he active
c o m p o n en t in lecithin for lobster nutrition w as identified as p h osphatidylcholine, since
ovine PE and soybean PI could not substitute fo r the effect o f soybean PC to reduce
m ortality in ju v en iles (D ’A bram o et al., 1981). Sim ilarly, K anazaw a (1993) reported a
gro w th -p ro m o tin g effect for Japanese flounder larvae only w hen soybean PC was
supplem ented, w hereas soybean PI clearly did not im prove grow th com pared to the
P L -free control diet. By contrast, soybean PI provided a h ig h er w eight gain for 70 days
old ayu than soybean o r bonito egg PC (K anazaw a et al., 1985b). F irst-feeding carp fed
8
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I 6
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o
5
4
1
0
0
1
2
3
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Fig. 2. G ro w th o f sh rim p postlarvae (P . vannam ei) as a function o f the level o f dietary PC , presented as
p u rified so y b ean PC (95% PC , O ) or de-oiled soybean lecithin (23% PC , • ) . D ata rep resen t dry w eight gain
after 41 d ay s o f cu ltu re from an initial w eight o f 3 mg (from C ou tteau et al., 1996).
41
156
P. C outteau e t al. / A q u a c u ltu re 155 (1997) 1 4 9 -1 6 4
a soybean P C -enriched diet e xhibited a h ig h er initial grow th rate but a h ig h er m ortality
rate, due to the ap p earance o f severe lordosis, than larvae fed a soybean P i-e n ric h ed diet
(G eurden et al., 1995a). T he differen t effects o f dietary PC and PI in the latter case o f
feeding fish larvae exclusively fioni first-feeding on artificial diets m erits further
investigation.
2.4. Influence o f the fa tty a c id com position o f the P L source
C hicken egg PC w as not effective in im proving grow th and survival o f larval P.
ja p o n ic u s, w hereas bonito-egg PC and soybean PC w ere effective (K a n azaw a et al.,
1985a). S im ilarly, lecithin and PC derived from soybean or bonito egg w ere m ore
effective in prom oting grow th o f larval ayu P. altivelis than c hicken e g g lecithin
(K anazaw a e t ah, 1981, 1985b). H ow ever, all these PL sources drastically im proved
grow th and survival o f the larvae com pared to the PL -deficient control, w hereas a
saturated species o f PC , dipalm itoyl PC , w as com pletely ineffective (K a n azaw a et al.,
1985b). Sim ilarly, D ’A bram o et al. (1981) found that the effectiveness o f PC -containing
PL to prevent m ortality in ju v en ile lobsters w as dependent on the con stitu en t fatty acids
w ith soybean PC being m ore effective than egg lecithin and dipalm itoyl PC. In addition,
1-acyl lyso soybean PL in diets fo r carp resulted in low grow th and survival, although
su p erio r to that o f the PL -free d iet (G eurden et al., 1995c). T he latter finding questions
the hypothesis that, in carp larvae, PL are absorbed after preferential hydrolysis at the
sn — 2 position as suggested in adult carp by Iijim a et al. (1990). In this w ay, carp larvae
w ould require intact p hospholipid m olecules containing sn — 2 unsaturated fatty acids
w ith e ith e r inositol or choline headgroups, w hich confirm s the presum ption o f K anazaw a
e t al. (1981, 1985b) for larval ayu.
2.5. Influence o f the p ro te in source in experim ental diets
Special attention should be paid to the poorly understood interaction betw een
ph ospholipid requirem ents and o ther dietary ingredients such as the protein source.
Juvenile A m erican lobsters fed sem i-purified diets based on casein show ed high levels
o f m ortality due to incom plete ecdysis (m olt death syndrom e, M D S), and this could be
allev iated by the supplem entation o f 7.5% o f dietary soybean lecithin (C onklin et al.,
1980). H ow ever, no PL requirem ent w as found fo r lobsters w hen purified crab protein
( < 0.02% total lipid), rather than casein, w as used as the prim ary protein source (K ean
e t al., 1985). M ore recently it w as show n that further purification o f the crab protein
co ncentrate by using 0.06M E D T A in a 50:50 isopropanol:w ater solvent m ixture
resulted in a reappearance o f a M D S for w hich soybean lecithin w as ineffective (C astell
et al., 1991; C onklin et a l , 1991). A lthough som e evidence was found by C astell et al.
(1991) that B vitam ins and m anganese m ay play a role in M D S induction, the
physiological factors responsible for m olt death rem ain to be elucidated. S im ilarly,
supplem entary lecithin in the diet w as not needed for larval P. ja p o n ic u s w hen a
m ixture o f casein and soybean protein w as used instead o f casein only (T eshim a et al.,
1993). Interestingly, the addition o f the lipids extracted from the soybean protein source,
con sistin g o f PC (37% ), L PC (23% ), and partial glycerides (19% ), w ere effective at
42
P. C outteau et al. / A q u a c u ltu re ¡55 ( 1997) ¡ 4 9 -1 6 4
157
concentrations as low as 0.43% o f the diet in im proving grow th and survival o f the
larvae fed the defatted soybean protein.
T h e possible interactions b etw een the selection o f the protein source and PL
requirem ents m ay have im p o rtan t im plications for the role o f lecithin in the form ulation
o f practical diets. F urtherm ore, the use o f practical diets for confirm ing PL requirem ents
detected with sem ipurified diets is poorly docum ented in literature. In this w ay, P.
m onodon ju v en iles fed a p ractical d iet containing approxim ately 1% o f e n d o g en o u s PL,
increased w eight gain significantly w hen 2% o f soybean lecithin w as added to the diet
(Piedad-P ascual, 1986). C om p arativ e studies using sim ilar PL sources added to both
c asein-based experim ental diets and practical diets based on shrim p a n d / o r fish m eal
w ould allow a better interpretation o f the present know ledge o f PL requirem ents.
3. Functionality of dietary phospholipids
3.1. Source o f m em brane p h o sp h o lip id s
Several authors have suggested that larval stages are not capable o f synthesizing PL
at a rate sufficient to m eet the requirem ents for the form ation o f new cell com ponents
during the initially short period o f rapid larval grow th (K anazaw a, 1993; G eu rd en et al.,
1995c). T he difference in the activity am ong PL, w ith PC generally being the m ost
active com pound, m ay then be ex p lain ed by the specific need for incorporation into
m em branes, w hich consist m ainly o f PC. T he present understanding o f the m etabolism
o f phospholipids in fish and sh rim p is m ainly based on extrapolations from the
know ledge o f m am m als (S a rg en t et al., 1993). It rem ains unclear w hether E FA , w hich
are preferentially esterified to the sn — 2 position in PL, are assim ilated as a single
entity o r separately from the PL m olecule. T he latter m ay have im portant im plications
for the possible direct e ffe c t o f the fatty acid com position o f dietary PL on the
com position and function o f tissue PL. C hen and Jenn (1991) found d ecreased n — 3
H U FA levels in the p olar fraction o f the m uscle lipids o f P. p enicillatus w ith increasing
dietary soybean PC, reflecting the high 18:2(n-6) level o f the PL source. T h e latter
authors suggested the presence o f a d ynam ic m etabolism in the m uscle PL pool in w hich
the PL are replaced by PL o f d ietary origin. In particular for m arine larvae, the
discrepancy betw een the fatty acid c o m position o f PL present in form ulated diets, w hich
are often from soybean origin, and that o f th eir natural food deserves further research.
3.2. P rovision o f choline, inositol, E F A a n d energy
Preferential catabolism o f PC during em bryonic and yolk sac stages has been
o bserved in various species o f m arine fish w ith the PC presum ed to be m ainly a source
o f inorganic phosphate and choline (herring: T o ch er et al., 1985), D H A (cod: F raser et
al., 1988) or m etabolic energy (cod, halibut, and plaice: R ainuzzo et al., 1992).
S im ilarly, it has been p ostulated that d ietary PL m ay serve as a direct source o f nutrients
for early feeding stages o f fish and crustacea.
43
P. C onneau et al. / A q u a c u ltu re 155 (1 9 9 7 i 1 4 9 -1 6 4
158
E ffect inositol v e r s u s PI
E ffect c h o lin e v e r s u s P C
8
14
7
12
6
10.
5
4
3
2
1
0
no PL c h o lin e E P C
n o PL inositol
SPI
Fig. 3. T heoretical biom ass (w eig h tX survival) o f larval c a rp (C. c a rp io ) fed dietary su p p lem en ts o f choline o r
inositol versus PC o r PI for 25 days after start-feeding. R esults are also show n for a P L -free control diet (no
P L). S ig n ifican t differences ( P < 0.05) are indicated w ith a d iffe re n t letter (from G eurden et al., I995a.c).
V arious form s and concentrations o f d ietary c holine w ere not as effective as lecithin
in red u cin g m olt death syndrom e in ju v en ile lobsters fed casein-based diets (C onklin et
al., 1980). Sim ilarly, the beneficial effect o f d ietary P C and PI on grow th and survival of
first-feed in g carp larvae could not be re p lica ted by the supplem entation o f choline or
inositol, respectively (G eurden et al., 1995a,c; Fig. 3), and the addition o f choline C D P
c o u ld n o t correct the PL deficiency in the d iet o f larval ayu (K anazaw a et al., 1985b).
N o n eth eless, this does not exclude the p o ssibility that PC can prevent a choline
d eficien cy as show n by H ung (1991) in a study w ith ju v en ile w hite sturgeon A cip en ser
transm ontanus.
In one o f th eir earlier w orks, K anazaw a e t al. (19 7 9 ) show ed that the grow th-prom o­
ting effects o f 1% dietary clam phospholipids on ju v e n ile P. ja p o n ic u s could not be
re p lica ted by including an equal level o f the fatty acids derived from this PL source in
the diet. Sim ilarly, the beneficial effects o f d ietary soybean lecithin on grow th and
survival o f larval ayu could not be attributed to the c ontent o f EFA , w hich was low in
so y b ean PL, w hereas EFA w ere supplied in su ffic ie n t am ounts in the diet (K anazaw a et
al., 1981). T he latter authors concluded that the g row th-prom oting effect o f PL was due
to c ertain effects o f the m olecular form o f the P L ra th e r than to the EFA provided by
them . F o r carp, sunflow er lecithin, very p o o r in n — 3 PU FA , provided better rearing
re su lts than did rapeseed lecithin or even a m arin e PL ex tract (G eurden et al., 1995c).
T h e fa c t th a t experim ental diets used to dem onstrate P L requirem ents are form ulated to
co n ta in sufficient am ounts o f E FA in the neutral lipid fraction (K anazaw a et al., 1981,
1985a,b; C am ara, 1994; G eurden et al., 1995a,b,c; C outteau et al., 1996) does not,
ho w ev er, exclude the possibility that pho sp h o lip id s m ay be superior to neutral lipids as
E F A source for larval stages, w hose digestive capacity m ay not be fully developed. PL
are m o re p o lar and m ay be m ore easily em u lsified and thus less susceptible to a
h y p o th etica l bile salt lim itation for their assim ilation (K oven et al., 1993; Sargent et al.,
1993). In this w ay, O lsen et al. (1991) suggested that larval and early juv en ile cod may
h a v e a n absolute need for polar lipids, both for the supply o f energy and for essential
fa tty acids, because o f the lim ited d igestibility o f neutral lipids.
44
P. C outteau et al. / A quaculture 155 (19971 1 4 9 -1 6 4
159
3.3. Im p ro ve m e n t o f d iet pro p erties
O ne hypothesis for the effectiveness o f dietary su p p lem en tatio n o f soybean lecithin in
preventing m olt death in lobsters fed casein-based diets is related to the reduction o f the
leaching o f w ater soluble nutrients, in particular m anganese and B vitam ins (C astell et
al., 1991). T he lim ited w ater stability o f m any sem ipurified diets that have been used to
study phosp h o lip id requirem ents in larval and postlarval stages should inspire further
e x p lo ra tio n o f this hypothesis in other species that have been reported to require PL. The
loss o f total dry m atter in casein-based m icrobound diets used fo r studies w ith postlarval
p enaeid shrim p am ounted to 40% after 10 m in o f exposure to w ater, and w as not
in flu en ced by the addition o f PL (C am ara, 1994). N evertheless, m ore detailed studies are
needed to evaluate the effect o f PL on the leaching rate o f m icronutrients.
P h ospholipids have been show n to exert antio x id an t (K ing et al., 1992; M cE voy et
al., 1995) and feed-attractant properties (H arada, 1987), and it rem ains to be investigated
w h eth er these m ay account to som e extent for the beneficial effects o f PL supplem enta­
tion to larval and ju v en ile diets.
3.4. P L em ulsifying properties
S everal authors have suggested that lecithin m ay be req u ired as a su rfactant for
e ffic ie n t lipid ém ulsification and digestion in early stages o f crustaceans and fish larvae
(K a n az a w a et al., 1979; C onklin et al., 1980; K oven et al., 1993). In this w ay, K oven et
al. (1 9 9 3 ) reported a 7-fold increase o f l4C -oieic acid in lipids o f 22-day-old Sparus
aurata w hen fed a lecithin supplem ented diet co m p ared to a PL -free diet. T hey
attrib u ted this increased lipid uptake to the em ulsifying function o f the lecithin.
H ow ever, various lines o f evidence indicate that d ietary PL im prove grow th and survival
by o th e r effects than the enhancem ent o f ém ulsification and absorption o f dietary lipid
in the d ig estiv e tract. T he addition o f taurocholic acid co u ld not substitute for the
b en eficial effect o f soybean PC either in larval P. ja p o n ic u s (K anazaw a et al., 1985a),
larval a y u P. altivelis (K anazaw a et al., 1985b), or in ju v e n ile H. am ericanus (D ’A bram o
no PL L-DSL
DSL C a-D SL
F ig. 4. T h e o retic al b iom ass (w eight X survival) o f larval carp (C . c a rp io ) fed d ietary supplem ents o f de-oiled
soy b ean lecithin (D SL ) w ith varying em ulsifying properties for 21 d ay s after start-feed in g . 1-acyl lyso D SL
(L -D S L ) an d C a2+-enriched D SL (C a-D S L ) have, respectively, h ig h e r and lo w er em ulsifying capability in
vitro than the untreated D SL. See also Fig. 3 for explanation (fro m G eu rd en el al„ 1995c).
45
160
P. C outteau et al. / A quaculture 155 (1997) 1 4 9 -1 6 4
et al., 1981). B esides, ju v e n ile P. ja p o n ic u s w ere capable o f assim ilatin g tripalm itin and
cholesterol efficien tly w ithout dietary PL (T eshim a et al., 1986c,d). In a recent study
w ith start-feeding carp larvae, it w as show n that PL added to a casein -b ased d iet as the
sole lipid source en h an ced grow th in the sam e w ay as w hen ad d ed to the sam e diet
supplem ented w ith 4% neutral lipids (G eurden et aí., 1995c). In the sam e study, it was
show n that the in vivo em ulsifying properties o f dietary soybean lecithin could not
explain the fish p erform ance: C a2+ enrichm ent o f the soybean lecithin, w hich decreases
its em ulsifying properties, did not affect its positive effect on gro w th and survival,
w hereas 1-acyl lyso lecithin, being a better o il/w a te r em ulsifier, g ave p o o re r culture
results than u ntreated soybean lecithin (Fig. 4).
3.5. R ole in lip id transport
V arious findings support the hypothesis that dietary PL e n h an c e the transport o f
lipids in crustaceans, i.e., the export o f absorbed lipids from the gut ep ith eliu m into the
haem olyrnph, and the m obility o f lipids betw een the various tissues and organs. T he lack
o f dietary lecithin in casein diets for ju v en ile lobster w as asso ciated w ith low er
haem olyrnph levels o f p h ospholipids and cholesterol (D ’A bram o et al., 1982). U sing
3H -cholesterol, D ’A bram o et al. (1985) found relatively h igher levels o f cholesterol in
the m idgut glan d and lo w er haem olyrnph levels in lobsters fed diets w ithout lecithin,
presum ably due to a decrease o f the transport rate o f cholesterol out o f the m idgut gland
into the hem olym ph. L o b sters fed lecithin-supplem ented diets ex h ib ited h igher levels o f
serum and lipoprotein cholesterol than those fed lecithin-deficient diets regardless of
w hether crab protein o r casein w as used as protein source (B aum e t al., 1990). T h e latter
finding show ed that the im pairm ent o f cholesterol transport due to a shortage of
lip o p ro te in -p h o sp h o lip id -c h o le ste ro l com plexes was not a possible cause o f m olt death
syndrom e observ ed in ju v e n ile lobsters fed casein diets, but co n firm ed the role of
lecithin in cholesterol transport. U sing radioactively labelled tripalm itin and cholesterol,
T eshim a e t al. (1986c,d) show ed that dietary PL im prove the m obilization o f cholesterol,
and to a lesse r extent, o f triglycerides from the gut to the hepatopancreas, hem olym ph
and m u scle in P. ja p o n ic u s. T he latter authors hypothesized that the dietary PL,
e specially PC , m ay act as an acyl donor for the lecith in x h o leste ro l acyltransferase w hich
converts free cholesterol into sterol ester (T eshim a et al., 1986d). In contrast to the
above sug g ested en h an cem en t o f cholesterol availability by d ietary PL, several re­
searchers failed to dem onstrate an interaction betw een lecithin and cholesterol require­
m ents (P . ja p o n ic u s : T esh im a et al., 1982; P. p e n ic illa tu s: C hen and Jenn, 1991; M.
rosenbergii: B riggs e t al., 1988). In larval fish, little is know n about the intervention o f
PL in lipid transport.
T esh im a e t al. (1986c) assum ed that dietary PL m ay provide specific lipid classes as
substrate for the form ation o f lipoproteins w hich are the m ain m ediators o f lipid
transport in the h em olym ph o f shrim p, and contain polar lipids as the m ain lipid
c o m ponent (T eshim a and K anazaw a, 1980). L ipoproteins also play an essential role in
lipid transport in the c irculatory system o f fish, and PC is the pred o m in an t p o lar lipid
class in fish lipoproteins (H en d erso n and T ocher, 1987; Sheridan, 1988). N evertheless, it
rem ains to be show n to w hat extent dietary PL supply m ay influence the pathw ays for
the synthesis o f lipoproteins responsible for the transport o f absorbed lipids.
46
P. C outteau e t al. / A quaculture 1 5 5 (1 9 9 7 ) 1 4 9 -1 6 4
161
3.6. E ffect o f P L on body lip id com position
T he inclusion o f PL in the d iet affected lipid deposition, resulting in in cre ased lipid
retention and levels in the anim al (T eshim a et al., 1986a; C hen and Jcn n , 1991,
T akeuchi et al., 1992). S om e con tro v ersy rem ains regarding the influence o f the level o f
dietary PL on the lipid class co m p o sitio n o f the anim al. Larval P. ja p o n ic u s feed in g on
a diet containing 3% soybean lecithin show ed higher tissue levels (% o f w et w eight) o f
SE, FS, PC and PI co m pared to larvae fed on a PL deficient diet (T eshim a et al., 1986e).
In a sim ilar study w ith ju v en ile P. ja p o n ic u s, T eshim a et al. (1986a) found increased
body levels o f PL, in p a rticu la r PC, and cholesterol due to the supplem entation o f 3%
soybean lecithin in the diet. Increased total lipid levels in hepatopancreas and hem olym ph
in ju v en ile P. ja p o n ic u s due to PL supplem entation w ere found to be due to an increase
o f both neutral lipids, i.e., triglycerides and cholesterol, and polar lipids, m ainly PC
(T eshim a et al., 1986b). In total m uscle lipid o f P. penicillatus. C hen and Jenn (1991)
did not observe a preferential increase o f the proportion o f PC in total lipids due to the
supplem entation o f purified PC up to 5% o f the diet.
An interaction betw een the incorporation efficiency o f dietary EFA and PL su p p le­
m entation has been d ocum ented in various species. T he proportions o f n-3 H U F A in
larval P. ja p o n ic u s varied w ith the type o f phospholipid supplem ented to the diet, w ith
the highest and low est p ro portion occurring in the larvae receiving soybean PC and
soybean PI, respectively (T esh im a et al., 1986e). A lso, a higher proportion o f E P A and
DFIA w as observed in ju v en ile P. ja p o n ic u s due to the addition o f 3% o f soybean
lecithin in the diet (T eshim a et al., 1986a). T he better efficiency o f dietary E FA due to
PL supplem entation is likely the basis for the interaction betw een E FA and PL
requirem ents, as w as dem o n strated by K anazaw a et al. (1985a) for larval P. ja p o n ic u s,
and deserves further investigation regarding its relevance in practical diets.
4. Conclusions
V arious experim ental results have show n the beneficial effects o f d ietary PL for
larval and ju v en ile stages o f several species o f fish and crustacea. N evertheless, further
research using purified diets and PL sources is still needed to identify the p hysiological
basis o f the requirem ent, and define the optim al dietary PL supply in term s o f PL classes
and PL fatty acid com position. Furtherm ore, the interaction o f PL req u irem en ts with
other nutrients such as the d ietary protein source and essential fatty acids d eserv es m ore
investigation.
Acknowledgements
T his w ork w as supported by the B elgian N ational Fund for S cientific R esearch
(N F W O ); the B razilian C ouncil fo r Science and T echnology (C N Pq); the F rench R egion
A quitaine; the Belgian M inistry for S cience Policy (O O A -Program m e); IN V E A quacul­
ture, B elgium ; V andem oortele, G roup R and D C enter, B elgium and L ucas M eyer,
B e lg iu m /G e rm a n y .
47
P. C outteau e t a i / A q u a c u ltu re J5 5 (1 9 9 7 ) 1 4 9 -1 6 4
162
References
B aum , N .A ., C onklin, D .E ., C hang, E .S., 1990. E ffect o f d ie tary lecithin in com bination w ith casein o r crab
protein on cholesterol uptake and transport in the lo b ste r H o m a ru s am ericanus. J. W orld A quacult. Soc.
21, 2 7 7 -2 8 7 .
B riggs, M .R .P ., Jauncey, K ., B row n, J.H ., 1988. T h e ch o lestero l an d lecithin requirem ents o f ju v e n ile praw n
( M acrobrachium rosenbergii) fed sem ipurified diets. A quacultu re 70, 1 2 1 -1 2 9 .
C am ara, M .R ., 1994. D ietary phosphatidylcholine req u irem en ts o f P enaeus ja p o n icu s Bate and P enaeus
va n n a m ei B oone (C rustacea, D ecapoda, P enaeidae). P h D T hesis. U niv. o í G hent, B elgium . 173 pp.
C astell, J., B oston, L.D ., C onklin, D .E., B aum , N .A ., 1991. N u tritio n ally induced m olt death syndrom e in
aquatic crustaceans: II. T he effect o f B vitam in and m an g an ese d eficiencies in lobster ( H om a ru s
am ericanus). C rustacean N utrition N ew sletter 7, 1 0 8 -1 1 4 .
C hapelle, S ., A bdul M alak, N ., Z ingelstein, G ., 1985. Inco rp o ratio n o f w -6 polyunsaturated fatty acids into
phospholipids o f the crab C arcinus m aenas. B iochem . S ystem . Eco!. 13, 4 5 9 -4 6 5 .
C hen, H .Y ., 1993. R equirem ents o f m arine shrim p, P e n a eu s m o n o d o n , ju v en iles for phosphatidylcholine and
ch o lestero l. A quaculture 109, 1 6 5 -1 7 6 .
C hen, H .Y ., Jenn, J.S., 1991. C om bined effects o f d ietary phosph atid y lch o lin e and cholesterol on the grow th,
.s u r v iv a l and body lipid com position o f m arine shrim p. P e n a eu s p en icilla tu s. A quaculture 96, 1 6 7 -1 7 8 .
C on k lin , D .E ., D evers, K., B ordner, C .E., 1977. D ev elo p m en t o f artificial diets for the lobster. H om a ru s
am ericanus. P roc. W orld M aricult. Soc. 8, 8 4 1 -8 5 2 .
C onklin, D .E ., D ’A bram o, L.R ., B ordner, C .E ., B aum , N .A ., 1980. A successful purified diet for the culture o f
ju v e n ile lobsters: the effect o f lecithin. A quaculture 21, 2 4 3 -2 4 9 .
C on k lin , D .E ., B aum , N .A ., C astell, J.D ., B oston, L .D ., H afang, L ., 1991. N utritionally induced m oll death
sy n d ro m e in aquatic crustaceans: I. Introduction to the problem . C rustacean N utrition N ew sletter 7,
1 0 2 -1 0 7 .
C o u tteau , P ., C am ara, M .R ., Sorgeloos, P ., 1996. T h e e ffec t o f differen t levels and sources o f dietary
phosphatidylcholine on the grow th, survival, stress resistance, and fatty acid com position o f postlarval
P en a eu s vannam ei. A quaculture 147, 2 6 1 -2 7 3 .
D ’A b ram o , L.R ., B ordner, C .E., C onklin. D .E ., B aum , N .A ., 1981. E ssentiality o f dietary phosphatidylcholine
fo r th e survival o f ju v e n ile lobsters. J. N utr. I l l , 4 2 5 - 4 3 1 .
D ’A bram o, L .R ., B ordner, C .E ., Conklin, D .E ., 1982. R elatio n sh ip b etw een dietary phosphatidylcholine and
seru m cholesterol in the lobster H om arus sp. M ar. B iol. 67, 2 3 1 -2 3 5 .
D ’A b ram o , L .R ., B aum , N .A ., B ordner, C .E., C on k lin , D .E ., C hang, E .S., 1985. D iet-dependent cholesterol
tra n sp o rt in the A m erican lobster. J. Exp. M ar. B iol. E col. 87, 8 3 -9 6 .
F raser, A .J., G am ble, J.C ., S argent, J.R ., 1988. C hanges in lipid co n ten t, lipid class com position o f developing
eggs o f co d ( G adus m orhua). M ar. B iol. 99, 3 0 7 -3 1 3 .
G eu rd en , I., C harion, N ., M arion, D ., B ergot, P ., 1995a. D ietary p h o spholipids and body deform ities in carp
C yp rin u s carpio L. larvae. In: L avens, P., Jaspers, E ., R o elan ts, I. (E ds.), Larvi ’95 — Fish and Shellfish
S y m p o siu m , G ent, B elgium . E urop. A quacult. S o c., S pec. P ub ., 24, 1 6 2 -1 6 5 .
G eu rd en , I., C outteau, P., Sorgeloos, P., 1995b. D ietary phospholip id s fo r European sea bass ( D icentrarchus
la b ra x L .) during first ongrow ing. In: L avens, P ., Jasp ers, E., R o elan ts, I. (E ds.), Larvi ’95 — F ish and
S h ellfish Sym posium , G ent, B elgium . E urop. A quacult. S oc., Spec. Puhi., 24, 175 -1 7 8 .
G eu rd en , I., R adiinz-N eto, J., B ergot, P., 1995c. E ssen tiality o f d ietary phospholipids for carp ( C yprinus
c a rp io ) larvae. A quaculture 131, 3 0 3 -3 1 4 .
H arad a, K ., 1987. R elationships betw een structure and feeding attractio n activity o f certain L-amino acids and
le cith in in aquatic anim als. N ippon Suisan G akkaishi 53, 2 2 4 3 -2 2 4 7 .
H en d erso n , R .J., T ocher, D .R ., 1987. T he lipid co m position and b iochem istry o f freshw ater fish. Prog. Lipid
R es. 2 6 , 2 8 1 -3 4 7 .
H ertram p f, J.W ., 1992. F eeding aquatic anim als w ith phospholipid s: II. Fishes. Lucas M eyer Publication No.
11. L u c as M eyer, H am burg, 70 pp.
H ilton, J.W ., H arrison, K .E ., Slinger, S.J., 1984. A sem i-purified test diet fo r M acrobrachium rosenbergii and
th e la ck o f need for supplem ental lecithin. A quaculture 37, 2 0 9 -2 1 5 .
H u n g , S .S.O ., 1991. N utrition and feeding o f hatch ery -p ro d u ced ju v e n ile w hite sturgeon ( A cip en ser transm on­
tanus): an overview . In: W illiot, P. (E d.), A cipenser. C E M A G R E F , pp. 6 5 -7 7 .
48
P. C onneau et al. / A q u a c u ltu re 155 ( 1997) 1 4 9 -1 6 4
163
H ung, S .S ., L utes, P .ß ., 1988. A prelim inary study on the n on-essen tiality o f lecithin for hatchery-produced
ju v e n ile w hite sturgeon ( A c ip en ser transm ontanus). A quacu ltu re 68, 3 5 3 -3 6 0 .
lijim a, N ., A ida, S., M ankura, M ., Kuyam a, M ., 1990. Intestinal ab so rp tio n and plasm a transport o f dietary
trig ly ce rid e and phosphatidylcholine in the carp (C yp rin u s ca rp io ). C om p. B iochem . P hysiol. 96A , 4 5 - 5 5 .
K anazaw a, A ., 1993. E ssential phospholipids o f fish and cru stacean s. In: K aushik, S .J., L uquet, P. (E ds.), Fish
N u tritio n in P ractice. Biarritz (F rance), June 2 4 -2 7 . Edn. IN R A , P aris 1993, Les C olloques nr. 61,
5 1 9 -5 3 0 .
K anazaw a, A .. K oshio, S.,
1994. Lipid nutrition o f the spin y lo b ster P a n u liru s ja p o n icu s (D ecapoda
P alin u rid ae): a review . C rustaceana 67, 2 2 6 -2 3 2 .
K anazaw a, A ., T eshim a, S., T okiw a, S., Endo, M .. A bdel R azek , F.A ., 1979. Effects o f short-necked clam
phosp h o lip id s on the grow th o f praw n. Bull. Jpn. Soc. S ei. F ish. 45, 9 6 1 -9 6 5 .
K anazaw a, A ., T eshim a, S., Inam ori. S., Iw ashita, T ., N agao, A ., 1981. Effects o f phospholipids on grow th,
survival rate and incidence o f m alform ation in the larval ayu. M em . Fac. F ish., K agoshim a U niv. 30.
3 0 1 -3 0 9 .
K anazaw a, A ., T eshim a. S., Inam ori, S., M atsubara, H ., 1983a. E ffects o f dietary phospholipids on grow th o f
the larval red sea bream and knife jaw . M em . Fac. F ish., K ag o sh im a U niv. 32, 1 0 9 -1 1 4 .
K anazaw a, A .. T eshim a. S., K obayshi, T ., T akae. M ., Iw ashita. T ., U ehara, R ., 1983b. N ecessity o f dietary
phosp h o lip id s for grow th o f the larval ayu. M em . Fac. Fish., K ag o sh im a U niv. 32, 1 1 5 -1 2 0 .
K anazaw a, A ., T eshim a, S., S akam oto. M ., 1985a. E ffects o f d ietary lipids, fatty acids, and phospholipids on
gro w th and survival o f praw n ( Penaeus ja p o n ic u s ) larvae. A q u acu ltu re 50, 3 9 - 4 9 .
K anazaw a, A ., T eshim a, S., S akam oto, M „ 1985b. E ffects o f d ie tary bo n ito -eg g phospholipids and som e
phosp h o lip id s on grow th and survival o f the larval ayu. P leco g lo ssu s a ltivelis. 2 . A ngew . Ichthyol. 4,
1 6 5 -1 7 0 .
K ean, J.C ., C astell, J.D ., B oghen, A .G ., D ’A bram o, L .R ., C on k lin , D .E ., 1985. A réévaluation o f the lecithin
and cholesterol requirem ents o f juvenile lobster ( H om arus a m erica n u s) using crab protein-based diets.
A q u acu ltu re 47, 1 4 3-149.
K ing, M .F ., B oyd, L .C ., Sheldon, B.W ., 1992. E ffects o f phosp h o lip id s o n lipid o x idation o f a salm on oil
m odel system . J. A m . O il Chem . Soc. 69, 2 3 7 -2 4 2 .
K oven, W .M ., K olkovski, S., T andler, A ., K issii, G .W ., S klan, D ., 1993. T h e effect o f dietary lecithin and
lipase, as a function o f age, on n-9 fatty acid incorporation in the tissu e lipids o f S p arus aurata larvae.
F ish P hysiol. Biochem . 10, 3 5 7 -3 6 4 .
L éger, P., B engtson, D .A ., S im pson, K .L., Sorgeloos, P ., 1986. T h e use and n u tritional value o f A rtem ia as a
food source. O ceanogr. M ar. B iol. A nn. Rev. 24, 5 2 1 -6 2 3 .
M cE v o y , L .S ., N avarro, J.C ., A m at, F., S argent, J.R ., 1995. O ptim isatio n o f a p h o spholipid-enhanced
en rich m en t em ulsion for A rtem ia. In: L avens, P ., Jasp ers, E., R o elan ts, I. (E d s.), L arvi ’95 — Fish and
S h ellfish S ym posium , G ent, B elgium . Europ. A quacult. S oc., S pec. P ubl., 24, 1 4 1 -1 4 4 .
O lsen, R .E ., H enderson, R .J., Pedersen, T ., 1991. T h e influen ce o f dietary lipid classes on the fatty acid
c o m p o sitio n o f sm all cod G adus m orhua L. ju v e n ile s reared in an en clo su re in northern N orw ay. J. Exp.
M ar. B iol. Ecol. 148, 5 9 -7 6 .
P ied a d -P a sc u al, F., 1986. E ffect o f supplem ental lecithin and lip id so u rces on the g row th and survival o f
P en a eu s m onodon ju v en iles. In: M acL ean, J.L ., D izon, L .B ., H o sillo s, L.V . (Eds.), T he F irst A sian
F ish erie s F orum , A sian F isheries Soc., M anila, P hilippines, pp. 6 1 5 -6 1 8 .
P o sto n , H .A ., 1990a. P erform ance o f rainbow trout fed supplem en tal so y b ean lecithin and choline. Prog. Fish
C u lt. 52, 2 1 8 -2 2 5 .
P o sto n , H .A ., 1990b. Effect o f body size on grow th, survival, an d ch em ical co m position o f A tlantic salm on
fed s o y lecithin and choline. Prog. Fish Cult. 52, 2 2 6 -2 3 0 .
P o sto n , H .A ., 1991. R esponse o f A tlantic salm on fry to feed-grad e lecithin and choline. Prog. Fish Cult. 53,
2 2 4 -2 2 8 .
R ad ü n z -N eto , J., Com aze, G ., C harlon, N „ B ergot, P., 1994. L ip id sup p lem en tatio n o f casein-based purified
die ts for ca rp ( C yprinus ca rp io L .) larvae. A quaculture 128, 1 5 3 -1 6 1 .
R ain u zzo , J.R ., R eitan, K.I., Jorgensen, L., 1992. C om parative stu d y on the fatty acids and lipid com position
o f fo u r m arine fish larvae. C om p. Biochem . Physiol. 103B, 2 1 - 2 6 .
R ain u zzo , J.R ., R eitan, K .I., O lsen, Y ., 1994a. E ffect o f short- and lo n g -term en rich m en t on total lipids, lipid
class and fatty acid com position in rotifers. A quacult. Int. 2, 1 9 -3 2 .
49
164
P. C outteau et a i / A quaculture 155 (1997) 1 4 9 -1 6 4
R ainuzzo, J.R ., R eitan, K .I., Jorgensen. L., O lsen, Y., 1994b. Lipid co m p o sitio n in tu rb o t larvae fed live feed
cu ltu red by em u lsio n o f d ifferent lipid classes. C om p. B iochem . P hysiol. 107A, 6 9 9 - 7 1 0 .
S argent, J.R ., B ell, J.G ., B ell, M .V ., H enderson, R .J., T o cher, D .R ., 1993. T h e m e tab o lism o f phospholipids
and poly u n satu rated fatty acids in fish. In: Lahlou, B., V itiello, P., (E ds.), A q u acu ltu re: F undam ental and
A pplied R esearch, C oastal and E stuarine Studies, 43. A m erican G eo p h y sical U nion, W ashington, pp.
1 0 3 -1 2 4 .
S heridan, M .A ., 1988. L ip id dynam ics in fish: aspects o f absorption, tran sp o rtatio n , dep o sitio n and m obiliza­
tion. C om p. B iochem . P hysiol. 90B , 6 7 9 -6 9 0 .
S hieh, H .S ., 1969. T h e biosynthesis o f phospholipids in the lobster H o m a ru s a m erica n u s. C om p. Biochem .
Physiol. 30, 6 7 9 -6 8 4 .
Sorgeloos, P ., C outteau, P., D hert, P., M erchie, G ., L avens, P., 1997. U se o f b rine sh rim p A rtem ia sp. in
larval cru sta cean nutrition: a review Rev. Fish. Sc., in press.
S zlam inska, M ., E scaffre, A .M ., B ergot, P., 1993. P relim inary data on sem isy n th etic diets for goldfish
( C arassius a u ra tu s L .) larvae. In: K aushik, S.J., L uquei P. (E ds.), Fish N u tritio n in P ractice, B iarritz
(F rance), June 2 4 - 2 7 . E d. IN R A , P aris 1993 (Les C olloques nr. 61), 6 0 7 -6 1 2 .
T ackaert, W ., C am ara, M .R ., S orgeloos, P., 1991. T h e effect o f d ietary p h o sp h atid y lch o lin e in postlarval
pen aeid shrim p. I. D iet preparation. In: L avens, P., S orgeloos, P., Jaspers, E., O lle v ier, F. (E d s.), Larvi '91
.— F ish and C rustacean L arviculture S ym posium . G ent, B elgium . E urop. A quacult. Soc., Spec. Pub!., 15.
7 6 -7 9 .
T akeuchi, T ., A rakaw a, T ., S atoh, S., W atanabe, T ., 1992. S u p p lem en tal effect o f pho sp h o lip id s and
req u irem en t o f eicosapentaenoic acid and docosahexaenoic acid o f ju v e n ile strip ed ja ck . N ippon Suisan
G akkaishi 58. 7 0 7 -7 1 3 .
T eshim a, S., K anazaw a, A ., 1980. Lipid constituents o f serum lipoproteins in the praw n . Bull. Jpn. Soc. Sei.
F ish. 4 6 , 5 7 -6 2 .
T eshim a, S ., K anazaw a, A ., S asada, H „ K aw asaki, M ., 1982. R eq u irem en ts o f larval praw n. Penaeus
ja p o n ic u s , for cholesterol and soybean phospholipids. M em . Fac. Fish., K ag o sh im a U niv. 31, 1 9 3 -1 9 9 .
T eshim a, S., K anazaw a, A ., K akuta, Y., 1986a. Effects o f dietary p h o sp h o lip id s o n g row th and body
co m position o f the ju v e n ile praw n. Bull. Jpn. Soc. Sei. Fish. 52, 1 5 5 -1 5 8 .
T eshim a, S ., K anazaw a, A ., K akuta, Y., 1986b. E ffects o f dietary ph o sp h o lip id s on lipid transport in the
ju v e n ile praw n. B ull. Jpn. Soc. S ei. Fish. 52, 1 5 9 -1 6 3 .
T eshim a, S., K anazaw a, A ., K akuta, Y ., 1986c. R ole o f dietary
tripaim itin in the praw n. Bull. Jpn. Soc.
pho sp h o lip id s in the tran sp o rt o f l4C
Sei. Fish. 52. 5 1 9 -5 2 4 .
T eshim a, S., K anazaw a, A ., K akuta, Y., 1986d. R ole o f dietary
ch o lestero l in the praw n. B ull. Jpn. Soc. Sei. Fish. 52, 7 1 9 -7 2 3 .
p h o spholipids in the transport o f MC
T eshim a, S ., K anazaw a. A ., K akuta, Y ., 1986e. G row th, survival and body lipid co m p o sitio n o f the praw n
larvae receiving several dietary phospholipids. M em . Fac. Fish., K ag o sh im a U niv. 35, 1 7 -2 7 .
T eshim a, S., K anazaw a, A ., H orinouchi, K ., Y am asali, S., H irata, H., 1987. P h o sp h o lip id s o f rotifer, praw n
an d larv al Fish. N ip p o n S u isan G akkaishi 53, 6 0 9 -6 1 5 .
T eshim a, S ., K anazaw a, A ., K oshio, S., 1993. R ecent developm ents in n u trition and m icroparticulate diets o f
larval praw ns. Israeli J. A quacult. 45, 1 7 5-184.
T ocher, D .R ., F raser, A .J., S argent, J.R ., G am ble, J.C., 1985. Lipid class com p o sitio n du rin g em bryonic and
ea rly larval dev elo p m en t in A tlantic herring ( C lupea harengus). L ipids 20, 8 4 -8 9 .
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Freshwater Biology (1997) 38, 501-512
Manipulation of dietary lipids, fatty acids and
vitamins in Zooplankton cultures
PE T ER C O U T T E A U * A N D P A T R I C K S O R G E L O O S
Laboratory o f Aquaculture & A rtem ia Reference Centre, U niversity o f Ghent, Rozier 44, B-9000 G hent, B elgium
*A u th o r to w hom correspondence should be sent
SUMMARY
1. A w ide range of species that are cultivated in commercial m ariculture are planktonic
during at least p art of their life cycle; for example, the larval stages of shellfish (shrimp
and molluscs) and the live feeds (rotifers, brine shrimp, copepods) used in the
larviculture of m arine fish and shellfish. Over the last decades various techniques have
been developed to deliver nutrients to these zooplanktonic organisms either through
artificial diets or by m anipulating the composition of the live prey fed to the
carnivorous stages. This paper reviews the methodology that has allow ed aquaculturists
to gain know ledge of nutritional requirements and m ay offer interesting opportunities
for ecologists to verify the importance of key nutrients in the natural food chain of
m arine as well as freshw ater ecosystems.
2. Live micro-algae can be replaced partially or completely in the diet of filter-feeders
such as rotifers, Artemia, shrim p larvae and bivalves, by various types of preserved
algae, m icro-encapsulated diets and yeast-based diets, whereas lipid em ulsions and
liposomes m ay be utilized to supplem ent specific lipid-and water-soluble nutrients,
respectively. M icrobound and micro-encapsulated diets have been designed to
supplem ent live feed in the culture of m icro-predators such as fish and shrim p larvae.
3. Live prey organisms, in particular rotifers and Artemia, can be 'bio-encapsulated' with
a variety of enrichm ent diets to m anipulate their content in certain nutrients, including
(»3 highly unsaturated fatty acids (FA) and the vitam ins C, A and E. Nevertheless, the
enrichm ent techniques are not applicable for all nutrients and prey organisms.
Phospholipid composition is difficult to m anipulate through the diet of live feed and the
enrichm ent of the essential FA docosahexaenoic acid (DHA) is ham pered in m ost
Artemia species due to the catabolism of this FA following enrichment.
Introduction
Commercial mariculture comprises a wide range of
species that are planktonic during at least part of their
life cycle, i.e. the larval stages of cultivated species
of fish and shellfish, and a few selected species of
Zooplankton (copepods, rotifers, brine shrimp) that
are used as live feed for the former. The natural diet
of these planktonic organisms, feeding either as filterfeeders or micropredators, consists of a wide diversity
of bacteria, detritus, phytoplarikton a n d /o r smaller
Zooplankton (Omori & Ikeda, 1994). This high
diversity of food organisms of different sizes and
© 1997 Blackw ell Science Ltd
biochemical composition provides good chances for
meeting all the nutritional requirements of the larvae.
Larviculture nutrition, more particularly for startfeeding early larval stages, appears to be one of the
major bottlenecks for the industrial upscaling of the
culture of many species of marine fish and shellfish.
Collecting and feeding natural plankton is not practical
on an industrial scale and natural food organisms are
not consistently available. Over the past decades, trial
and error approaches have resulted in the adoption
of selected larviculture diets, various species of
501