Aquaculture, 79 (1989) 157-161 157

Elsevier Science Publishers B.V., Amsterdam - Printed in The Netherlands

The Effect of Enrichment Diets on the Fatty Acid

Composition of the Rotifer Brachionus plicatilis

JOSE RODRIGUEZ RAINUZZO’, YNGVAR OLSEN’ and GRETHE ROSENLUND’

‘University of Trondheim, Laboratory of Biotechnology, N-7034 Trondheim (Norway)

‘SINTEF Division of Applied Chemistry, Center of Aquaculture, N-7034 Trondheim (Norway)

ABSTRACT

Rodriguez Rainuzzo, J., Olsen, Y. and Rosenlund, G., 1989. The effect of enrichment diets on the
fatty acid composition of the rotifer Brachionusplicatilis. Aquaculture, 79: 157-161.

Cultures of the rotifer Brachionus plicatilis (200 ind./ml, 0.5 ,ug dry weightlind.) pre-fed on
yeast and capelin oil (1O:l w/w) were given different enriched diets and harvested after 24 h. The
fatty acid composition of the rotifers was affected by that of the feed. The highest level of n-3
highly unsaturated fatty acids (HUFA) (38% ) and 22:6 n-3 ( 19% ) was obtained after enrichment
of the rotifers fed with squid meal. A positive relation was found between the n-3 HUFA in the
enrichment diet and that of the rotifers (P -=0.05, t-test). The cultivation feed of the rotifers may
nevertheless be more important for the n-3 HUFA level than the enrichment feed. Starvation for
49 h did not change the fatty acid composition of the rotifers.

INTRODUCTION

Since Japanese workers (Ito, 1960) pioneered the use of the rotifer Bru-
chionus plicatilis as a live feed for the rearing of small fish larvae, much infor-
mation on the nutritional quality of rotifers has appeared (Watanabe et al.,
1978,1983; Imada et al., 1979; Kitajama et al., 1979; Ben-Amotz et al., 1987;
Dendrinos and Thorpe, 1987; Lubzens et al., 1987). Several reports have dem-
onstrated that the content of highly unsaturated fatty acids (HUFA) of the n-
3 series in rotifers is an important factor for their nutritional value for marine
fish larvae.
Baker’s yeast, Saccaromyces cereuisiue, supplemented with small amounts
of fish oil may be used as cultivation feed for rotifers. Their nutritional value
may be improved by feeding a high quality diet shortly before they are to be
used as larval feed (Watanabe et al., 1978; Ben-Amotz et al., 1987).
In the present study we investigated the effect of different artificial enrich-
ment diets on the fatty acid composition of the rotifer Brachionus plicatilis.

0044~8486/89/$03.50 0 1989 Elsevier Science Publishers B.V.

158

MATERIALS AND METHODS

In the first experiment the rotifers were cultured in a ZOO-1tank (20’ C and
21 ppt salinity). The animals were fed once daily with baker’s yeast and capelin
oil (1O:l w/w) and cultured to a density of approximately 200 ind./ml (100 pg
dry wt/ml). The culture was distributed into 10 vessels (20 1) and thereafter
given different artificial diets as shown in Table 1.
The artificial feed (2 g) and 200 ml of sea water were homogenized in a
blender for 10 s and added to the vessels, which then were transferred to a
refrigerated room (7-8°C). After 24 h the rotifers were collected on 90-ym
plankton netting, thoroughly washed (20 ppt sea water) and immediately
frozen.
In another experiment a culture of rotifers (200 ind./ml) fed baker’s yeast
and capelin oil for several weeks, was starved for 49 h. Samples were collected
after 24 and 49 h and treated as above.
The total lipids were extracted by the method of Bligh and Dyer (1959), and
calculated on a dry weight basis. The fatty acids were isolated after alkaline
hydrolysis (0.5 N NaOH) and methylated with 12% BFB in methanol (Met-
calfe et al., 1966). The fatty acid methyl esters were determined by capillary
gas chromatography (Carlo Erba HRGC 5160 series equipped with a SP-2330
glass capillary column, splitless injection and flame ionization detector). The

TABLE 1

Total lipid content ( f SE*) of the diets and in rotifers fed for 24 h on the diet

Diet Enrichment diets Total lipid content ( % of dry weight)

no.
Feed Rotifers

1 Baker’s yeast + capelin oil 2.1kO.l 9.4kO.2

2 Baker’s yeast 2.2kO.l 9.OkO.2
3 Baker’s yeast + cod liver oil N.A. 11.8&0.3
4 Squid meal** 11.9f0.2 10.5 + 0.1
5 Shrimp meal** 3.1 kO.1 8.3 f 0.2
6 Herring roe meal** 15.0+0.1 9.1 f 0.1
7 Fish meal** 11.7kO.2 8.7fO.l
8 Mixed diet l** (containing squid meal,
fish meal, vitamins and minerals) 9.9kO.l 8.1_+ 0.1
9 Mixed diet 2** (containing shrimp meal,
fish meal, vitamins and minerals) 9.7+0.1 8.1 f0.2
10 Mixed diet 3** (containing squid meal,
herring roe meal, vitamins and minerah) 11.7fO.l 8.7fO.l

N.A., no date available.

*Standard error of the mean.
**go-pm micronized.
 159

concentrations were estimated by a Shimadzu Chromatopac C-R3A computing

integrator. The identification was based on standard references (Nu-Chek
Prep., Elysian, MN, U.S.A.) and graphical procedures (Ackman, 1969).

RESULTS

The rotifers had a rather constant total lipid content (range &l-1,1% ) after
24 h feeding on different diets, even though the lipid content of the diets varied
widely (range &l-15%). The groups fed on baker’s yeast and cod liver oil (3)
and on squid meal (4) had the highest levels of total lipids (Table 1).
The contents of n-3 HUFA in the feed and in the rotifers after 24 h feeding
are shown in Figs. la and b, respectively. The lipids in herring roe meal (6)
contained the highest amount of n-3 HUFA (48% ) , whereas baker’s yeast did
not contain such fatty acids at all. The variation of n-3 HUFA in rotifers was
less than that in the feed. The rotifers fed on squid meal (4) showed the highest

1 2 3 4 5 6 7 8 9 10

E
: 40 [3 22:6 n-3
;
P
9 3o
u
fj 20
e

$ 10

E
0
1 2 3 4 5 6 7 8 9 10

Fig. 1. (a) n-3 HUFA content (% of total fatty acids) of different enrichment diets; (b) n-3 HUFA
content(% of totalfatty acids) in rotifers fed for 24 h on different enrichment diets. Numbers
indicate the diets (cf. Table 1) .
160

40r
A4

A n-3 HUFA

0 22:6 n-3

0 10 20 30 40 50

Fig. 2. n-3 HUFA and 22:6 n-3 content (% of total fatty acids) in 24-h enriched rotifers as a
function of the respective content of the enrichment feed. Numbers indicate the diets (cf. Table
I).

level of n-3 HUFA (38%) even though squid meal had a lower n-3 HUFA
content than the herring roe meal (6). The content of 22:6 n-3 in the rotifers
showed the same pattern of variation as the n-3 HUFAs, with the highest level
in squid meal (19% ).
The results revealed a significant (P~0.05) positive relationship between
the n-3 HUFA and 22:6 n-3 content in the diet and that of the rotifers (Fig.
2).
No essential changes in the fatty acid composition of rotifers occurred when
the rotifers were starved for 49 h at 16 oC. The content of lipids was, however,
reduced by 25% during the phase of starvation.

DISCUSSION

The results revealed that the fatty ac?d composition of enriched rotifers was
dependent on the fatty acid composition of the feed. The 24-h enrichment
procedure applied in this study turned out to yield predictable fatty acid com-
position and content of n-3 HUFA in the rotifers, although the data were some-
what scattered (Fig. 2). This may be due to the availability of the dietary par-
ticles for the rotifer as well as to the com&ition and digestibility of the lipids
in the different diets.
The rotifers maintained their relative n-3 HUFA content through 49 h star-
vation, as has also been found by Lubzens et al. (1985 ) . This suggests that the
feed used for cultivation of the rotifers prior to enrichment may be important
for the content of n-3 HUFA in the rotifers after enrichment, as well as the
enrichment diet. This is believed to be important if the n-3 HUFA content of
 161

the enrichment diet is lower than that of the cultivation feed, or if the culti-
vation diet is rich in n-3 HUFA.
This study suggests some possible enrichment diets that may be useful to
improve the nutritional value of rotifers as a food for marine fish larvae.

ACKNOWLEDGEMENTS

We thank John Korstad, Olav Vadstein and Marte Morkved for their advice
and assistance. This study forms part of a research project on live feed for
marine fish larvae financed by the Royal Norwegian Council for Scientific and
Industrial Research.

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