Aquaculture 160 1998.

89102

Effect of microalgal and inert cornmeal and

cornstarch / diets on growth performance and
biochemical composition of
Ruditapes decussatus seed
A. Perez
Camacho
a

a,)

b
, M. Albentosa a , M.J. Fernandez-Reiriz
,

b
U. Labarta

Instituto Espanol
S N, 15080 A Coruna,
de Oceanografa,
Muelle de Animas
Spain
b
Instituto de Inestigacions
Marinas,
Eduardo Cabello 6, 36208, Vigo, Spain
Accepted 20 October 1997

Abstract
Research was carried out into the effect of phytoplankton, cornmeal and cornstarch diets on
growth and biochemical composition of the seed of the little-neck clam, Ruditapes decussatus.
The seed of R. decussatus, fed on daily rations of Isochrysis galbana organic weight. of 0.5 and
1% of live weight of the seed, showed an improvement in growth rate when cornstarch, which is
99% carbohydrate, was added to these diets. Thus in the case of a daily ration of 0.5%, daily
growth rates increased by between 33.5 and 32.3%, depending on whether we are referring to
organic weight, dry weight or live weight, when 1.5% cornstarch was added. In the case of a
ration of 1% I. galbana, the addition of another 1% cornstarch lead to an improvement in daily
growth rates, depending on the different weight class in question, of between 14.1 and 15.5%.
When compared to a daily ration consisting of 2% phytoplankton, which was considered to be the
optimal ration for growth in the seed of these clams, the replacement of half the quantity of I.
galbana by a quantity of cornstarch of equivalent weight gave a growth rate in terms of organic
weight of 87.9% that of the phytoplankton diet, while the rates for dry weight and live weight
were 89.6 and 87.9%, respectively. These results improved noticeably when cornmeal, consisting
of 10% protein and 90% carbohydrate, was used instead of cornstarch. In the case of a 2%
phytoplankton diet, if we substituted an equivalent quantity of cornmeal for 50% of the

Corresponding author. Fax: q34-81229077; e-mail: alejandro.perez@co.ieo.es.

0044-8486r98r$19.00 q 1998 Elsevier Science B.V. All rights reserved.

PII S 0 0 4 4 - 8 4 8 6 9 7 . 0 0 2 3 2 - 9

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A. Perez
Camacho et al.r Aquaculture 160 (1998) 89102

phytoplankton, the growth rate in organic matter was the same 99.0%. as those for the diet
consisting of phytoplankton alone, while growth rates in dry weight and live weight were 6.2 and
5.9% higher, respectively, than those of the phytoplankton diet. It would therefore appear that
cornmeal and to a lesser extent cornstarch. can be successfully used as a partial substitute for
phytoplankton in diets for the seed of R. decussatus and its use in hatcheries and nurseries
devoted to the culture of this species would lead to a considerable reduction of production costs.
q 1998 Elsevier Science B.V.
Keywords: Ruditapes decussatus; Seed culture; Inert food; Cornmeal; Cornstarch

1. Introduction

The little-neck clam, Ruditapes decussatus, is one of the most important species in
Spanish aquaculture, commanding high prices in the market. Possibilities for production
are limited due to the scarcity of naturally-occurring seed and the high cost of seed
production in commercial hatcheries. Any saving in this cost is therefore crucial for the
viability of this type of industry and would be a major contribution to the expansion of
aquaculture production.
The basis for the development of the bivalve mollusc culture industry was established
at the mid-point of this century in works by Quayle 1952.; Loosanoff and Davis 1963.
and Walne 1956., among others. Ever since then the production of live phytoplankton
has been seen as one of the major contributory factors to the costs borne by hatcheries
and nurseries Persoone and Claus, 1980; De Pauw, 1981; Lucas and Gerad, 1981.,
accounting for 30% of the total seed production cost in the former Coutteau and
Sorgeloos, 1992. and up to 85% of production costs in the latter Bolton, 1982..
Various authors have attempted to find a substitute for live phytoplankton in the diet
of bivalve molluscs and have experimented, among other products, with spin-dried and
freeze-dried algae Laing et al., 1990; Laing and Gil Verdugo, 1991; Laing and Millican,
1992., yeasts Epifanio, 1979; Urban and Langdon, 1984; Nell, 1985., modified yeasts
Albentosa et al., 1989; Coutteau et al., 1991., microcapsules Langdon and Waldock,
1981; Jones et al., 1984; Langdon and Bolton, 1984; Langdon et al., 1985; Laing, 1987.
and different varieties of cereal starch Haven, 1965; Ingle, 1967; Dunathan et al., 1969;
Castell and Trider, 1974; Wisely and Reid, 1978; Nell and Wisely, 1983, 1984; Langdon
and Bolton, 1984; Langdon and Siegfried, 1984; Urban and Kirchman, 1992.. In none of
these studies is R. decussatus mentioned and their results vary considerably according to
the characteristics of the foodstuff and the species for which it is intended; until now, no
total substitute has been found for live phytoplankton, the best results having been
obtained with 50% substitution or less.
The purpose of this study is to assess the performance of cornmeal and cornstarch in
the diet of the seed of the little-neck clam R. decussatus, whether in diets exclusively
composed of these products or in mixed diets with varying proportions of phytoplankton. Seed growth rates and biochemical composition were used to assess the results
obtained from the different diets.

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91

2. Materials and methods

2.1. Seed
The experiments were carried out with seed of R. decussatus obtained from
broodstock conditioned in the Instituto Espanol
de Oceanografa.
The methods used to
induce spawning and to cultivate the larvae and the seed were those described by Perez

Camacho et al. 1977..

2.2. Diet
The basal diet consisted of the microalga Isochrysis galbana, which was replaced in
varying proportions by cornmeal or cornstarch. Preliminary trials were carried out to
define the sedimentation rates of the corn products concerned in the culture vessels and
equivalent amounts of the products were then added to the daily ration to compensate
for losses caused by sedimentation.
The microalgae were grown in 6-l glass flasks in a temperature-controlled chamber,
at a constant temperature of 188C and with constant illumination at 9900 lux. Salinity
was maintained at a constant 33. The culture medium used was that described by
Walne, 1956 and the microalgae were harvested in the initial stationary growth phase.
The cornstarch contained 99% carbohydrate, while the cornmeal contained 90%
carbohydrate and 10% protein. Both products were suspended in seawater by shaking
and sieved through a 20 m m sieve. A Coulter Counter Multisizer was used to measure
the concentration of the foodstuff in the seawater.
2.3. Experimental design
The base diet for the different experiments was an optimal daily ration of 2% organic
seed weight of the microalga I. galbana Albentosa et al., 1996.. The experiments were
carried out in triplicate for each diet. Each replica contained an initial biomass of 200
mg of clams live weight..
2.3.1. Experiment 1
This experiment was used to compare the growth rates achieved with the basal diet
F100, consisting of a daily ration of 2% I. galbana, with the two mixed diets M50 and
M25, in which 50% and 75%, respectively, of the phytoplankton was replaced by an
equivalent weight of cornstarch. Three control diets were established: F50, consisting of
a daily ration of 1% I. galbana; F25, consisting of a daily ration of 0.5% I. galbana; and
CS, consisting of 2% cornstarch. Percentages are expressed in terms of the live weight
of the clams. Initial figures for the seed were as follows: mean length, 2.10 . 0.20 mm;
mean live weight, 2.46 . 0.04 mg; mean dry weight, 1.58 . 0.04 mg; mean organic
weight, 0.20 . 0.01; and mean organic matter 12.5 . 0.1%. The experiment lasted 5
weeks.

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Camacho et al.r Aquaculture 160 (1998) 89102

2.3.2. Experiment 2
Cornmeal was used in place of cornstarch in this experiment. Four diets were tested,
consisting of varying proportions of phytoplankton and cornmeal, as follows: Diet F100,
2% I. galbana; diet M50, 50% phytoplankton and 50% cornmeal; diet M25, 25%
phytoplankton and 75% cornmeal; and diet CM, 2% cornmeal. The seed used in this
experiment was of similar size to that used in experiment 1 mean lengths 1.76 . 0.16
mm, mean live weights 1.34 . 0.06 mg, mean dry weights 0.85 . 0.04 mg, mean
organic weights 0.12 . 0.01, mean organic matters 13.2 . 0.1%.. The experimental
period was 3 weeks.
2.4. Experimental conditions
The seed was placed on the bottom of 6-l plastic vessels which were fitted with an air
point in order to reduce the sedimentation rates of the foodstuff. The experimental
cultures were kept in a temperature-controlled chamber at a constant temperature of
208C. The water in the vessels was changed daily, using seawater filtered through a 1
m m sieve and sterilised by UV. The experimental period was from 3 to 5 weeks.
The cornmeal and cornstarch were suspended in seawater, in 6-l round-bottomed
flasks with strong aeration in order to keep the food in suspension. The food was
supplied over 15-min periods at 4-h intervals by means of a 12-channel Ismatec MV-CA
pump. Each diet was assayed in triplicate and a fourth culture vessel was used to
determine the sedimentation rate.
The initial volume in the culture vessels was 3-l. In order to avoid the production of
pseudofaeces the concentration of the food in the vessels was kept below 1.6 m g mly1
of organic matter Albentosa et al., 1996.. The initial volume of water in the culture
vessels was increased weekly, according to the increase in biomass of the clams during
the same period, in order to maintain constant conditions throughout the experiment.
2.5. Diet assessment parameters
The live weight LW. of the clams in each experimental batch was obtained weekly,
after a 10-min draining period on absorbent paper. At the end of the experimental period
half of the clams in each batch were used to obtain the length of the seed L., the dry
weight DW., the weight of inorganic matter IW., the organic weight OW s DW y IW.
and the percentage of organic matter %OM s OWrDW= 100.. DW was determined
by drying the clams at 1008C for 24 h, while IW was obtained by heating to 4508C for
12 h. The other half of the batch was used for biochemical analysis.
The daily growth rate DGR. was calculated according to the equation
)

DGR: LnW1 y LnW0 . rt . 100

where W1 and W0 are the values of the different variables OW, DW, LW and L. at the
end and beginning, respectively, of each experiment and t is the number of days.

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93

2.6. Analytical methods

The organic content of the phytoplankton was determined by filtering the algal
culture through Whatman GFrC glass-fibre filters which had previously been rinsed and
ashed at 4508C. The filters were rinsed in a 0.5 M ammonium formate solution to
eliminate salt residues, dried to a constant weight and ashed at 4508C. The organic
content and water of the corn products were determined at 1008C and 4508C respectively.
The method described by Lowry et al. 1951. was used to determine the protein
content, following alkaline hydrolysis with NaOH 0.5 Nr24 hr308C. Total carbohydrates were quantified as glucose by the phenolsulphur method Strickland and
Parsons, 1968.. Lipids were extracted by a modification of the method of Bligh and
Dyer 1959. Fernandez-Reirz

et al., 1989. as follows: lipids were extracted by means

of chloroform:methanol 1:2. and then centrifuged, after which the sediment was once
again extracted with chloroform:methanol 2:1.. The resulting extract was purified by
rinsing both supernatants in a mixture of chloroform, methanol and water 8:4:3. as
described by Folch et al. 1957.. Total lipids were gravimetrically determined through
evaporation of the solvent in the purified extract on aluminium sheets at 60808C.
2.7. Statistical methods
The statistical package Statgraphics was used to analyse the results. For each
experiment growth was compared by examining the daily increase for each of the
variables listed above, while the daily growth rate DGR. was used to compare the
results of different experiments. Comparison between the different variables used to
evaluate the diets was performed by an ANOVA with a significance level of P - 0.05.
Angular transformation was used for values expressed as percentages and logarithmic
transformation for the comparison of weight increases. The Bartlett test was used to test
for variance homogeneity. Multiple comparisons were carried out with the Newman
Keuls multiple range test Snedecor and Cochran, 1971; Zar, 1974..

3. Results
3.1. Cornstarch
3.1.1. Experiment 1
When the daily diet of 2% I. galbana diet F100. was reduced to 1% diet F50., the
growth rate of the seed of R. decussatus decreased to 43% of that obtained with diet
F100 and decreases still further to 24% if the diet was reduced to 0.5% phytoplankton
diet F25..
If cornstarch, which is 99% carbohydrate Table 1., was added to diets F50 and F25,
growth rates increased considerably. Thus, in the case of the 0.5% ration, DW growth
increased by 80.1% when 1.5% cornstarch was added, while OW increased by 55.0%,

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94

Table 1
Mean daily increase in organic weight OW., dry weight DW., live weight LW. and shell length L., and %
organic matter OM. of clams fed on different diets average values of 3 replicates"stand. error.. Diet F100:
2% I. galbana OW. of clam LW daily; diet F50: 50% F100; diet F25: 25% F100; diet M50: 50% F100 and
50% cornstarch; diet M25: 25% F100 and 75% cornstarch; diet CS: 2% cornstarch OW. of clam live weight.
Initial figures: mean length, 2.10.0.20 mm; mean live weight, 2.459.0.041 mg; mean dry weight,
1.578.0.038 mg; mean organic weight, 0.198.0.005; and mean organic matter 12.5.0.1%.

F100
F50
F25
M25
M50
CS

OW m g indy1 .

DW m g indy1 .

LW m g indy1 .

L mm.

OM %.

45.3"4.8
19.5"0.4
10.9"0.4
31.4"1.5
16.9"0.9
2.6"0.9

375.7"21.9
201.4"0.7
102.7"6.3
263.6"6.1
185.0"0.9
46.4"2.4

593.8"33.2
321.9"11.1
170.9"7.0
433.9"12.4
288.4"2.6
65.3"3.6

85.1"0.9
59.2"2.1
38.9"3.1
70.1"1.0
55.4"1.3
12.7"1.0

12.6"0.7
10.2"0.1
10.6"0.3
12.0"0.4
9.8"0.4
10.2"0.4

Table 2
Mean daily increase in organic matter OW., dry weight DW., live weight LW. and shell length L., and %
organic matter OM. of clams fed on different diets average values of 3 replicates"SE.. Diet F100: 2% I.
galbana in OW. daily with regard to clam live weight; diet M75: 75% F100 and 25% cornmeal; diet M50:
50% F100 and 50% cornmeal; diet M25: 25% F100 and 75% cornmeal; diet CM: 2% cornmeal in OW. with
regard to clam live weight. Initial figures: mean length, 1.76.0.16mm; mean live weight, 1.341.0.059 mg;
mean dry weight, 0.854.0.039 mg; mean organic weights 0.119.0.005; mean organic matter, 13.2.0.1%.

F100
M75
M50
M25
CM

OW m g indy1 .

DW m g indy1 .

LW m g indy1 .

L mm.

OM %.

17.6"1.2
20.6"1.0
17.5"2.8
8.6"1.2
1.8"0.3

131.0"1.0
173.0"4.1
151.3"4.3
89.4"5.1
26.2"3.1

207.6"2.4
261.3"2.7
235.6"4.7
136.8"2.9
28.1"2.7

57.6"0.6
69.0"1.0
61.4"1.0
43.3"2.0
4.8"1.1

14.1"0.3
12.1"0.2
12.0"0.5
10.9"0.7
10.7"0.3

LW by 68.8% and L by 42.4%. For 1% rations of I. galbana, when a 1% supplement of

cornstarch was added, DW increased by 30.9%, OW by 61.0% and L by 18.4%.
In comparison with diet F100, when half the quantity of I. galbana was replaced by
an equivalent weight of cornstarch diet M50. the growth rate for OW was 69.3% of that
obtained with a phytoplankton only I. galbana. diet; DW was 73.7%, LW was 73.1%
and L was 82.4%. A 25% phytoplanktonr75% starch diet diet M25. gave growth rates
of 37.4, 51.7, 48.6 and 65.1% for OW, DW, LW and L, respectively, when compared to
the growth rates obtained with diet F100.

Fig. 1. Variation in daily growth rates of organic matter, dry weight, live weight and seed length for the seed
of the little-neck clam R. decussatus fed different proportions of phytoplankton, cornstarch A. and cornmeal
B.. Initial figures: mean length, 2.10.0.20 A. and 1.76.0.16 mm B.; mean live weight, 2.459.0.041 A.
and 1.341.0.059 mg B.; mean dry weight, 1.578.0.038 A. and 0.854.0.039 mg B.; mean organic
weight, 0.198.0.005 A. and 0.119.0.005 mg B.; mean organic matter, 12.5.0.1 A. and 13.2.0.1% A..
Vertical lines represent 95% confidence intervals.

A. Perez
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A. Perez
Camacho et al.r Aquaculture 160 (1998) 89102

The ANOVA gave significant differences P - 0.001. between daily growth rates for
OW, DW, LW and L. The NK test detected significant differences P - 0.05. in OW,
DW, LW and L between all groups with the exception of diets F50 and M25, where
there was no significant difference P ) 0.05..
Where the %OM of the seed is concerned, the NK test showed two homogeneous
groups, one formed by diets F100 and M50, with a %OM of between 12.0 and 12.6%
and the other by diets F50, F25, M25 and CS, with a %OM ranging from 9.8 to 10.6%.
There were significant differences between the two groups P - 0.05..
3.2. Cornmeal
3.2.1. Experiment 2
The growth rates of clams on inert diets obtained in experiment 1 were considerably
improved if we substitute cornmeal, which consisted of 90% carbohydrate and 10%
protein, for cornstarch Table 2.. Thus, compared with a base daily ration of 2% organic
weight of I. galbana with respect to the live weight of the clams diet F100., clams fed
on a diet consisting of 75% phytoplankton and 25% cornmeal diet M75. achieved OW
growth rates of 116.6% compared to those of the basal diet, while growth rates for DW,
LW and L were 32.1, 25.9 and 19.8% higher, respectively, than those corresponding to
the basal diet Table 2.. When 50% of the basal diet was replaced by cornmeal diet
M50., OW growth rates were the same 99.3%. as those for the diet composed of
phytoplankton only and growth in DW, LW and L were higher by 15.5, 13.6 and 6.6%,
respectively.
When the proportion of phytoplankton was reduced to 25% and the remaining 75%
replaced by cornmeal diet M25., the growth rates for OW, DW, LW and L were 48.6,
68.2, 65.9 and 75.2%, respectively, of the growth rates for diet F100.
The ANOVA showed significant differences between the diets P - 0.001. and the
NK test gave significant differences P - 0.05. between DW and LW among all the
diets, while the only differences for L which were not considered significant P ) 0.05.
were those between diets F100 and M50. In the case of OW, the differences between
diets F100 and M50, as well as those between M50 and M75, were not significant.
Diet had a significant effect on the percentage of organic matter in the seed
ANOVA, P - 0.001., although this effect was not the same as that on growth. Thus the
highest %OM corresponded to diet F100, followed by M75, M50 and M25, with the diet
consisting exclusively of cornmeal showing the lowest %OM. The NK test detected
significant differences P - 0.05. between all diets, except between M75 and M50, as
well as between CM and M25, where the differences were not significant.
3.3. Comparison of cornstarch and cornmeal diets
The use of cornmeal in place of cornstarch in mixed diets had a notably positive
effect on growth rates of the seed of R. decussatus. Thus, with cornstarch diet M25 the
daily growth rate DGR. in OW was 3.9, as opposed to 4.5 for the cornmeal diet M25
Fig. 1., which was significantly different ANOVA, P - 0.05.. Equally significant was
the difference in DGR between the two different M50 diets, which gave values of 6.7
for cornmeal and 5.3 for cornstarch.

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97

So positive was the effect on growth when cornstarch was substituted for cornmeal
that, as can be seen in Fig. 1, the DGRs for the M25 cornmeal diet were comparable to
those of the F50 phytoplankton and M50 cornstarch diets, with no significant differences
between them when compared by an ANOVA P ) 0.05.. The differences between diets
Table 3
Biochemical composition %. of the different cornstarch and cornmeal diets used in the experiments
Biochemical composition of diets %.
Diets

Protein

Carbohydrate

Lipid

Diets

Protein

Carbohydrate

Lipid

F100
M25
M50
M75
CM

33.4
16.1
21.9
27.7
10.3

30.9
74.3
59.8
45.3
88.8

35.7
9.6
18.3
27.0
0.9

F100
F50
F25
M25
M50
CS

33.4
33.4
33.4
8.7
17.0
0.5

30.9
30.9
30.9
82.3
65.1
99.4

35.7
35.7
35.7
9.0
17.9
0.1

Table 4
Cornstarch experiments. Biochemical composition m g indy1 and %. of the clams at the beginning and at the
end of the experimental period
Initial biochemical composition of clams
Protein m g indy1 .

Carbohydrate m g indy1 .

Lipid m g indy1 .

106.5"10.1

53.2

53.7"12.4

26.8

39.9"10.9

19.9

Final biochemical composition of clams

Diets
Protein m g indy1 .
%
Carbohydrate m g indy1 .

Lipid m g indy1 .

F100
F50
F25
M25
M50
CS

31.1
34.0
27.0
32.3
35.3
48.0

298.1"73.3
123.3"17.1
83.2"11.0
137.4"72.0
226.0"70.0
37.3"12.0

16.7
13.9
14.2
17.4
17.4
12.8

931.0"64.6
460.8"23.9
343.3"10.9
398.3"42.4
615.1"18.0
114.0"11.0

52.2
52.1
58.8
50.3
47.5
39.2

555.3"60.3
300.6"40.3
157.9"8.0
255.2"42.2
458.0"13.0
139.5"24.1

Table 5
Cornmeal experiments. Biochemical composition of the clams at the beginning and at the end of the
experimental period m g indy1 and %.
Initial biochemical composition of clams
Protein m g indy1 .

Carbohydrate ( m g indy1 .

Lipid m g indy1 .

54.4"4.1

46.1

37.8"3.9

32.0

25.8"2.5

21.9

Final biochemical composition of clams

Diets
Protein m g indy1 .
%
Carbohydrate m g indy1 .

Lipid m g indy1 .

F100
M25
M50
M75
CM

31.5
31.4
32.6
33.4
44.7

99.0"11.6
60.4"6.2
98.7"27.1
104.4"5.5
20.8"2.3

20.2
20.2
20.3
18.9
14.1

236.6"30.4
145.2"7.1
229.4"12.0
262.9"15.0
61.0"4.3

48.3
48.4
47.1
47.7
41.2

154.4"15.9
94.1"6.6
159.1"17.7
184.3"8.2
66.2"4.4

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Camacho et al.r Aquaculture 160 (1998) 89102

CS and CM, consisting wholly of cornstarch and cornmeal, were not significant
P ) 0.05. and neither were those between the F100 diets in experiments 1 and 2.
3.4. Biochemical composition
Tables 35 show the biochemical composition of each of the diets studied in
experiments 1 and 2, as well as the initial and final biochemical composition of the
clams fed these diets. As can be seen in these tables, the degree to which food was
assimilated depended to a large extent on the initial biochemical composition of the
clams: in experiment 1, for example, carbohydrate content was low at the beginning of
the experiment and this component was therefore stored, as an energy reserve, to a
greater extent than were lipids, thus giving rise to a relatively lower percentage of the
latter in the total content. The composition of the diet only produced major variations in
the biochemical composition of the clams in the case of extremely unbalanced diets such
as those consisting exclusively of cornmeal and cornstarch. In these cases the clams
maintained their initial quantities of protein and lipid and only assimilated carbohydrate,
leading to a relatively large increase in the proportion of this component and a
corresponding decrease in the proportions of protein and lipid.

4. Discussion
The results of the present study demonstrate that cornmeal and to a lesser extent
cornstarch, constitute a good complement to live phytoplankton in diets for the seed of
the little-neck clam R. decussatus. Cornstarch has been used to complement natural food
in order to improve the condition index of adult oysters Haven, 1965; Ingle, 1967;
Dunathan et al., 1969., thus emphasizing the importance of the accumulation of reserves
in the form of glucogen in gametogenesis Gabbott, 1976.. The use of artificial diets for
juvenile oysters containing carbohydrate in the form of starch has also been described
Castell and Trider, 1974; Wisely and Reid, 1978; Langdon and Bolton, 1984, Langdon
and Siegfried, 1984; Nell and Wisely, 1984; Urban and Langdon, 1984.. Growth rates
achieved with these diets were on the whole lower than those for natural diets.
Other authors have examined freeze-dried algae, dried- heterotrophically grown algae
and various microparticles as substitutes for live phytoplankton. The results of these
studies vary widely: freeze-dried and dried-heterotrophically grown phytoplankton can
only be used as partial substitutes for live phytoplankton, with very similar production
costs Curatolo et al., 1993; Laing et al., 1990; Laing and Gil Verdugo, 1991, Laing and
Millican, 1992., while microcapsules, apart from their very high cost, suffer from the
disadvantages of sedimentation, bacterial degradation, leaching of nutrients and poor
digestibility Langdon and Bolton, 1984; Chu et al., 1987..
Epifanio 1979., using diets consisting of 50% dried yeast Candida utilis and 50%
phytoplankton, achieved growth rates in length and dry weight of soft tissue comparable
to those of a 100% algal ration for Argopecten irradians, Mytilus edulis and Mercenaria mercenaria, while on the other hand the growth rate of Crassostrea irginica was
in inverse proportion to the quantity of yeast in the diet. Albentosa et al. 1989., working

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99

with R. philippinarum and Coutteau et al. 1994. with M. mercenaria, found that
replacing 50% of the algal ration with manipulated yeast did not result in a significant
decrease in growth rate relative to the algal-fed controls and the substitution of 80% of
the algal diets resulted in growth rates reaching 90%. Experiments carried out in our
laboratories have shown that the behaviour of R. decussatus resembles that of C.
irginica described above, since growth rates for mixed diets of yeast and phytoplankton
were considerably lower than those for both the phytoplankton control diets and the
cornstarch and cornmeal diets described in the present study.
In this study, the replacement of 50% phytoplankton by cornmeal leads to DGRs for
DW that were comparable to those for purely phytoplankton diets and when 75%
phytoplankton was replaced by cornmeal, DGRs for DW of 67% those of the control
diet were attained. When the results are expressed in terms of LW, the DGRs for the
M50 and M25 diets are 106 and 81%, respectively, of the DGRs for the control diet,
which underlines the importance of determining the variation in OW in this kind of trial,
since it provides a better indication of the quality of the diet than LW alone.
The results of our study, together with those on the use of wheatgerm Fernandez
Reirz
et al., 1998., are the only ones which consider the use of artificial diets for R.
decussatus. According to these results, cornmeal gives growth rates that are comparable
to those of the best quality artificial diets that have been examined to date, with the
added advantage of ease of handling and lower cost, which are common to all cereal
flours. If we consider that at present live phytoplankton is the usual food for broodstock
and seed of molluscs in bivalve hatcheries and nurseries and its production accounts for
30% of total seed production cost in the former Coutteau and Sorgeloos, 1992. and up
to 85% of the cost of production in the latter Bolton, 1982., the use of cornmeal as a
partial substitute for phytoplankton would lead to a considerable reduction of production
costs in those hatcheries that produce R. decussatus.
No clear definition of the nutritional requirements of bivalves is forthcoming from the
various studies that are available. Enright et al. 1986. achieved good growth rates in
juvenile Ostrea edulis with diets containing 15.5% protein and 59.4% carbohydrate,
while Wisely and Reid 1978. and Nell and Wisely 1983, 1984., in their study of
artificial diets for the fattening of Saccostrea commercialis, indicate that the optimal
level of carbohydrate and protein in the diet were 74 and 18%, respectively, while lipid
levels of around 10% total organic matter of the diet were sufficient to ensure optimal
indices of condition. On the other hand, Kreeger and Langdon 1993. achieved optimal
growth rates in juvenile Mytilus trossulus with diets that contain over 40% protein.
A comparison between the M50 diets in experiments 1 and 2 of this study, which
both contained similar amounts of lipid, showed that an increase in protein from 17 to
22% leads to a large increase in DGR. It may equally be true that the lower growth rate
for the M25 diet in experiment 2 compared to that of the M50 diet in experiment 1 is
related to the greater lipid content of the latter diet 18% as opposed to 9%., since both
diets contained a similar amount of protein, while postlarvae and see in the first months
after metamorphosis have a high lipid requirement Holland and Hannant, 1974.. The
results of experiment 2 show that a substitution of up to 50% phytoplankton for
cornmeal allows growth rates similar to those of wholly phytoplankton diets to be
achieved, suggesting that in the case of the seed of R. decussatus minimum protein and

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A. Perez
Camacho et al.r Aquaculture 160 (1998) 89102

lipid requirements are of the order of 22% and 18%, respectively Table 5.. The fact that
there are only minimal differences between diets CM and CS, composed entirely of
cereals, in spite of the higher protein content of the former, may indicate a lack of
essential amino acids in these proteins, together with an almost total absence of lipid in
both diets.

Acknowledgements
We would like to thank C. Fernandez
Pena, J. Daz,

L. Nieto and B. Gonzalez

for
their helpful technical assistance. This study was funded by CICYT-IEO-CSIC project
I q D number AGFC-1003-CO2-01.

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