HELGOL.

~NDER MEERESUNTERSUCHUNGEN
Helgol6nder Meeresunters. 43, 171-181 (1989)

E f f e c t s of d i e t o n p o p u l a t i o n d e v e l o p m e n t of t h e r o t i f e r
Brachion us p l i c a tiffs in c u l t u r e
M. Planas 1 & A. Est6vez 2
1 Instituto de Investigaciones Marinas, CSIC; Muelle de Bouzas 6, 36208 Vigo, Spain
2 Granja Atl~ntica de Couso S.A.; 15960 Riveira, La Corufia, Spain

ABSTRACT: Experiments were conducted in order to observe the effect of five diets on the
population development of the rotifer Brachionus plicatilis M/lller under laboratory conditions. Diets
were based on baker's yeast (Saccharomyces cerevisiae) and the algae Tetraselmis suecica and
Isochrysis galbana, mixed, or as simple diets. Growth rates, fecundity and biometric parameters
were studied for 15 days. The cultures were divided in a logarithmic phase and a harvesting phase.
Rotifers fed on Tetraselmis, alone or mixed with yeast or Isochrysis, gave good performances with
the best results in all the parameters studied. Average growth rates in all diets were similar during
the exponential phase, with values ranging from 0.72 (Tetraselmis and Tetraselmis + yeast) to 0.47
(yeast). During the harvesting phase there were high differences between diets, with rates highly
reduced in the yeast-group (0.17) and good rates when Tetraselmis was ingested (0.65-0.51). This
alga had a positive influence on the rotifersl increasing individual growth and fecundity.

INTRODUCTION

Following the first e x p e r i m e n ts carried out by Ito (1960) on cultures of Brachionus

plicatilis Mfiller and their initial application in a q u a c u l t u r e (Theilacker & McMaster,
1971; Howell, 1973), this species has n o w b e c o m e a prey of prime importance in the food
chain for raising a large variety of marine fish larvae. A l t h o u g h there has recently b e e n a
t e n d e n c y to suppress or, at least, to r e d u c e live food in the diet, b r o u g h t about by the
d e v e l o p m e n t of t e c h n i q u e s for p r o d u c in g microcapsules, rotifers remain an indispensible
prey in E u r o p e a n marine aquaculture.
The semicontinuous culture of rotifers b e g i n s with an initial inoculation of a small
quantity at low density in controlled conditions of light, t e m p e r a t u r e and salinity
(Theilacker & McMaster, 1971; D e v a u c h e l l e & Girin, 1974; Amat, 1975; Person le Ruyet,
1975; Hirata, 1979; Yfifera & Pascual, 1980; G a t e s o u p e & Luquet, 1981; Giliberto &
Mazzola, 1981), using yeast or a lg a e as a food source. H a r v e s t i n g occurs w h e n culture
density and v o l u m e are a d e q u a t e .
Although, initially, cultures of B. plicatilis w e r e fed exclusively on algae, the high
cost of al g ae production (Helm & Laing, 1981) initiated the use of b a k e r ' s yeast (Sac-
charomyces cerevisiae). H o w e v e r , using yeast as an only food source sometimes gener-
ated substantial d e c r e a s e s in production rates, so s u p p l e m e n t a r y food sources h ad to be
given, notably algae. T h e two algae c o m m o n l y used in Europe are Tetraselmis suecica
and Isochrysis galbana. Certain research p a p e r s h a v e u s e d both species an d yeast as the

9 Biologische Anstalt Helgoland, Hamburg

172 M. Planas & A. Estdvez

food source for B. plicatilis (Person le Ruyet, 1975; Scott & Baynes, 1978; Hirata, 1979;
Hirata & Mori, 1967; Yfifera & Pascual, 1980), but n o n e has c o m b i n e d all three. Existing
d a t a therefore m a k e a comparison b e t w e e n the three food sources difficult. In this
investigation w e h a v e b r o u g h t all t h r e e t o g e t h e r to establish their effect, s i n g l y or in a
m i x e d diet, on the population d e v e l o p m e n t of B. plicatilis u n d e r i d e n t i c a l e x p e r i m e n t a l
conditions. Furthermore, given that in fish farms rotifer cultures a r e h a r v e s t e d d u r i n g
rotifer m a s s production, we h a v e i n c o r p o r a t e d in our study a h a r v e s t i n g p h a s e in o r d e r to
collect d a t a r e g a r d i n g exploitation, since the latter a s p e c t is missing in the existing
studies on B. plicatilis p o p u l a t i o n d e v e l o p m e n t .

MATERIAL AND METHODS

The e x p e r i m e n t was carried out using 15 t r a n s p a r e n t cylindrical v e s s e l s e a c h with a
c a p a c i t y of 2 litres, in an isotherm c h a m b e r (20 ___ 1 ~ with constant a e r a t i o n , illumina-
tion (daylight fluorescent l a m p s p r o v i d i n g 60 BE m -2 sec -1, 24 hours) a n d salinity (34 %o).
A p o p u l a t i o n of Brachionus pficatilis from the Instituto Espafiol de O c e a n o g r a f i a in
Vigo was u s e d a n d prior to the c o m m e n c e m e n t of the test was m a i n t a i n e d in s t a r v e d
conditions for 24 hours in order to minimize the effects of previous diet.
The e x p e r i m e n t w a s b e g u n with 500 ml in e a c h vessel with a d e n s i t y of 20 rotifers/ml
(10 000 rotifers p e r vessel) a n d 21.8 % of e g g - c a r r y i n g females. The s e a w a t e r u s e d h a d
previously b e e n sterilised by p a s s i n g it t h r o u g h an ultraviolet light unit.
The 15 vessels w e r e d i v i d e d into 5 groups (3/group) e a c h one of w h i c h w a s fed with
the following nutritional sources a n d doses:
Group T: Tetraselmis suecica (0.75 x 106 cls/ml)
Group I: Isochrysis galbana (2 x 106 clslml)
Group Y: Yeast (Saccharomyces cerevisiae) (0.15 g/l)
Group YT: Yeast (0.075 g/l) and T. suecica (0.375.><106 cls/ml)
Group TI: T. suecica (0.375 x 106 cls/ml) and [. galbana (1 • 106 cls/ml)
The algae c a m e from cultures in exponential growth, were maintained at 20 ~ and
fed the Walne medium. Each day the n u m b e r of rotifers/ml, ovigerous females and the
n u m b e r of eggs carried were counted (5 counts of 1 ml each). The algae cells were
counted daily (i0 countings using a haematocytometer slide) to maintain a set concen-
tration of algae in each vessel and the quantity of algal culture which h a d to be added
w a s determined according to the following formulae:
- T, I and Y T groups:

V A = V0 " (CI - Co)I(CA - CI)

VA: volume of algae to be added; CA: density (cls/ml) of algae culture to be used (for
example: 0.75 • 106 cls/ml in group T); Co: density of algae in each vessel (algae not
ingested by rotifers);Vo: actual volume of rotifer culture in each vessel.

- TI group (mixed algal diet):

Vo 9 (C1 - Col) V2 " C2

VI= + ~
CA1 - C1 CA2
 Effects of diet on population d e v e l o p m e n t of Brachionus plicatilis 173

Vo 9 (C2 - Co2) Vl 9 C1
V2 = + - -
CA2 - C2 CA1
Vl a n d V2: volume of algae (species 1 a n d 2) to b e a d d e d ; Vo: actual v o l u m e of rotifer
culture in each vessel; C1 and C2: p r e d e t e r m i n e d density of a l g a e (species 1 a n d 2) in
e a c h vessel; CA1 a n d CA2: density of algal cultures (species 1 a n d 2) to b e used.
Yeast was a d d e d s u s p e n d e d in sterihzed water.
From d a y 7, w h e n the m a x i m u m volume of 2 litres w a s reached, to d a y 14, w h e n the
e x p e r i m e n t ended, cultures were h a r v e s t e d b y r e m o v i n g half the rotifers a n d half the
v o l u m e of each vessel each day. T h e n e a c h vessel was refilled up to 2 htres with sterilized
w a t e r after the addition of the daily food dose.
On d a y s 1, 4, 7, 10 a n d 13, 50 rotifers of each group w e r e r e m o v e d a n d fixed with
l u g o r s iodine; b o d y (lorica length) a n d e g g sizes w e r e m e a s u r e d .
Using the density m e a s u r e m e n t s of rotifer o b t a i n e d in the culture, the intrinsic
g r o w t h rate (K) a n d the doubling time (Td) w e r e c a l c u l a t e d both during the e x p o n e n t i a l
p h a s e (days 3-7) a n d the exploitation p h a s e (days 8-14), using the following equations:

In Nn+ 1 - In N n w h e r e Nn: density in d a y n (tn)

K=
tn + t -- tn Nn + 1: density in d a y n + 1 (tn + a)

In 2 0.693
Td . . . .
K K
K values during the exploitation p h a s e w e r e o b t a i n e d considering Nn as the density
of rotifers after r e m o v a l of half the volume a n d Nn+ 1 as the density before exploitation.
Analysis of variance (two-way ANOVA) was p e r f o r m e d to d e t e r m i n e the influence of
diet on the population development, using the daily K r e a d i n g s for e a c h vessel, - diet a n d
a g e of the culture b e i n g fixed factors (Sokal & Rohlf, 1979).

RESULTS
T h e c h a n g e s in the population, e x p r e s s e d as the total n u m b e r of rotifers, are shown
in F i g u r e 1.
In each of the 5 groups a l a g - p h a s e was o b s e r v e d which e n d e d on d a y 3 w h e n the
e x p o n e n t i a l p h a s e b e g a n , During the e x p o n e n t i a l p h a s e (days 3 to 7) the b e s t growths
w e r e o b s e r v e d in groups T (21.39 x 104 Rot.) a n d YT (18.55 x 104 Rot.), whilst the worst
growth w a s p r e s e n t in group Y (7.79 x 104 Rot.). The h i g h e r a n d lower densities o b t a i n e d
in this p h a s e w e r e 105 rot/ml (T-group) a n d 46 rot/ml (Y-group) w h e n the 2-htres volume
was r e a c h e d (day 7).
The calculation of the intrinsic growth rate a n d the d o u b l i n g time (Fig. 2 a n d Table 1)
s e r v e d to establish a comparison with the d a t a o b t a i n e d b y other authors as well as to
quantify the p o p u l a t i o n d e v e l o p m e n t in the c o n v e n t i o n a l manner.
Rotifers fed on y e a s t n e e d more t h a n two days to double their p o p u l a t i o n w h e r e a s
those fed on Tetraselmis only require slightly m o r e than one day, w h e t h e r fed on
Tetraselmis alone or c o m b i n e d with a n o t h e r food. Isochrysis fails b e t w e e n the two.
Growth c h a n g e s in the same w a y during the e x p o n e n t i a l phase, in that the five groups
m a i n t a i n a very similar a v e r a g e rate (lower in group Y) v a r y i n g b e t w e e n 0.72 a n d 0.47. In
174 M . P l a n a s & A. E s t 6 v e z

Rot. (104)/vessel

20-

18.

16-

12.

81 YT
TI
61 T

,i
21
1 2 3 4 5 6 7 8 9 10 11 12 13 14 Day

Fig. 1. Rotifer population (Rot. 104/vessel __ S.D. on day 7) in the five groups (Y: yeast; I: Isochrysis;
TI: Tetraselmis + Isochrysis; YT: yeast + Tetraselmis; T: Tetraselmis). Significant differences
b e t w e e n m e a n s on days 7 a n d 14 (**" p < 0.001)

Table I. M e a n g r o w t h rates (K) a n d d o u b l i n g (Td) in this study a n d those o b t a i n e d by: (a) Yfifera &
Pascual, 1980; (b) Scott & Baynes, 1978; (c) H i r a y a m a & W a t a n a b e , 1973; ": m a r i n e y e a s t + Chlorella
(1:1); (d) J a m e s et al., 1983; yeast + Chlorefla. (Y: yeast; Ii Isochrysis; TI: Tetraselrnis + Isochrysis;
YT: y e a s t + Tetraselmis; T: Tetraselmis)

Day Y I TI YT T

K 0.47 0.65 0.63 0.65 0.72

3-7
Td 1.47 1.07 1.12 1.07 0.96
K 0.17 0.34 0.65 0.58 0.51
8-14
Td 4.08 2.04 1.07 1.19 1.36

0.29 0.45 0.64 0.61 0.59

K 0.20a 0.43b 0.21a 0.42a
0.36c 0.64c*
3-14
2.39 1.54 1.08 1.13 1.17
Td 3.46a 1.62b 3.30a 1.65a
1.92c 1.08c ~
1.4-5.9d
 Effects of diet on population d e v e l o p m e n t of Brachionus plicatilis 175

1,0'

0.8,
TI
YT
0.6'

0.4'
T
0.2.

O.

-0,2.

-0.4.

-0.6

2 3 4 5 6 7 8 9 10 11 12 13 14 Day

Fig. 2. Rotifer growth rates (K) in the five groups (Y: yeast; I: Isochrysis; TI: Tetraselrnis + Isochrysis;
YT: yeast + Tetraselrnis; T: Tetraselmis)

this phase, d o u b l i n g times are quite similar with values r a n g i n g from 0.96 d a y s (T group)
to 1.47 days (Y-group). During the exploitation p h a s e (days 8-14), only the g r o u p that
receive Tetraselmis suecica maintain or i m p r o v e the growth rate of the previous phase,
group Y d r o p p i n g to a m e a n value of 0.17, clearly insufficient to support exploitation.
After onset of exploitation there is a g r a d u a l decline in the population size d e p e n d i n g on
the type of food given (Fig. 1). This decline is most noticeable in groups I a n d Y whilst in
the others there is a h i g h e r resistance to exploitation.
Table II shows the results o b t a i n e d from the analysis of variance. A level of
significance of p < 0.01 was o b t a i n e d w h e n differences of dietary effects on p o p u l a t i o n
d e v e l o p m e n t during the e x p o n e n t i a l p h a s e w e r e studied. T h e s e differences w e r e g r e a t e r
still during the exploitation p h a s e (p < 0.001). The s a m e significance level w a s o b t a i n e d
for the a g e factor during the e x p o n e n t i a l growth but it was lower (p < 0.01) during
harvesting. The interaction b e t w e e n diet a n d a g e of the p o p u l a t i o n does not s e e m to
affect the growth rate during the e x p o n e n t i a l p h a s e but its influence i n c r e a s e s d u r i n g the
harvesting p h a s e (p < 0.01). This interaction during the exploitation p h a s e can be
e x p l a i n e d as the p r e s e n c e of a different p o p u l a t i o n structure in the five groups as a result
of exploitation (rejuvenation) a n d different fecundities d e p e n d i n g on diet quality.
Thus, during the e x p o n e n t i a l growth phase, diet and a g e factors act i n d e p e n d e n t l y ,
with K values following a parallel path in the five groups (Fig. 2) although the r e a d i n g s
are h i g h e r in some than in others. However, w h e n half the volume of the cultures is
r e m o v e d daily the influence of a g e is not so important as an individual factor, b u t this
I76 M. Planas & A. Est4vez

Table 2. Two-way ANOVA from rotifer growth rates (K) during the exponential phase (days 3-7) and
the exploitation phase (days 8-14) (" : p < 0.05; ** : p < 0.01; * * *: p < 0.001)

Days 3-7 g.1. S.S. M.S. F

Diet 4 0.404 0.101 3.87" *

Age 3 4.256 1.419 54.37 ***
Diet x Age 12 0.461 0.038 1.46 N.S.
Error 40 1.045 0.026

Days 8-14 g.1. S.S. M.S. F

Diet 4 3.132 0.783 22.97 *" *

Age 6 0.859 0.143 4.19""
Diet x Age 24 1.796 0.075 2.20*"
Error 70 2.386 0.034

factor continues to h a v e an i m p o r ta n t influence in conjunction with the t y p e of nutritional

source.

Reproduction

Figures 3a and 3b give the daily fecundity rates. The greatest n u m b e r of e g g s carried
p er ovigerous f e m a l e per day (Fig. 3a) are 2.81 for group YT, 2.66 for g r o u p TI, 2.57 for
group T, 2.12 for group I and 2.09 for group Y, falling on day 4 of t h e experiment.
T h r o u g h o u t the experiment, groups T, YT an TI h a v e a n a v e r a g e n u m b e r of 2 e g g s w h i l e
groups I and Y h a v e a significantly l o w e r fecundity.
T h e daily fecundity rate for the population as a whole, i.e. the n u m b e r of e g g s p er
individual, w h e t h e r mature or i m m a t u r e (Fig. 3b) r each es a p e a k on d a y 3 w h e n the
values rise a b o v e 1 in all cases, doubling the population and t r i g g e r i n g t h e e x p o n e n t i a l
phase. Th e rate then drops, but less in groups T, YT and TI, with n e w p e a k s ev er y 3 days.
Growth rates show 3-day cycles w h i c h a g r e e with the e g g production cycles e x c e p t in the
Y-group (compare Figs 2 a n d 3: days 6, 7, 9, 11 and 13).

Biometry
We have studied the variation in sizes of both rotifers and eggs on 50 individuals per
group (days I, 4, 7, 10 and 13). Significant differences (p < 0.001) in body length of
rotifers according with the diet administered were observed although the difference
between the largest size (T) and the smallest (Y) was only 7.85 %. Rotifers fed on diets
including Tetraselmis were nearly 220 ~m long. The smallest individuals were those fed
on yeast (204 ~m). The influence of the nutritional source on the ovigerous female length
and on egg size was also significantly different (p < 0.001), both being greater when T.
suecica was ingested.
 Effects of d i e t o n p o p u l a t i o n d e v e l o p m e n t of Brachionus plicatilis 177

E / ~ ovig.
3. a

TI
YT
2. T

1 I

I ...... "'" .... .. ":

1.5.

1.0.
u

0.5-

I +
n.s. 0,t.m

; 2 3 4 5 6 7 8 9 1'0 1'1 1"2 1'3 14 da

Fig. 3. Rotifer fecundity rates (Y: yeast; I: Isochrysis; T[: Tetraselmis + Isochrysis; YT: yeast +
Tetraselmis; T: Tetraselrnis). (a) eggs carried per ovigerous female, ,(b) eggs carried per individual
(F-test in both fecundity-a-and-b-on days 7 and 14: * * * p < 0.001; n.s.: non significant)

F r o m F i g u r e 4, w h i c h s h o w s t h e v a r i a t i o n i n l e n g t h d i s t r i b u t i o n w i t h r e s p e c t to t i m e ,
the following deductions can be made:
- Initial size of t h e i n d i v i d u a l s fairly h i g h ( 2 1 8 - 2 3 0 ~ m o n d a y 1)
- A p p e a r a n c e of a h i g h p r o p o r t i o n of y o u n g r o t i f e r s o n d a y 4, a c h a r a c t e r i s t i c of t h e
b e g i n n i n g of t h e a c c e l e r a t i o n p h a s e
- P r e s e n c e of a m o r e h o m o g e n e o u s p o p u l a t i o n o n d a y 7 w i t h a g r e a t e r p r o p o r t i o n of
adults than on day 4
- R e j u v e n a t i o n of t h e p o p u l a t i o n s o n d a y 10 e x c e p t i n TI. In t h e Y - g r o u p t h e p r e s e n c e of
i m m a t u r e s p e c i m e n s is v e r y n o t i c e a b l e
- O n d a y 13 t h e r e is a b a l a n c e d p o p u l a t i o n o n l y w h e n 7+. suecica is u s e d i n t h e diet.
178 M, Planas & A. E s t f v e z
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 Effects of diet on population d e v e l o p m e n t of Brachionus plicatilis 179

DISCUSSION
In the e x p e r i m e n t a l conditions adopted, the p o p u l a t i o n a n d biometric p a r a m e t e r s
show the positive effect of Tetraselmis suecica, w h e t h e r fed singly or in a m i x e d diet, on
Brachion us plica ~flis.
The initial u n d e r n o u r i s h m e n t of the rotifers due to the starvation of the i n o c u l a (to
minimize the effects of the previous diet) b r o u g h t a b o u t a fairly long l a g - p h a s e of 2-3
days (2 days according to Yfifera & Pascual, 1984). The l a g - p h a s e l e d to a p e a k of e g g s
p e r individual, which m e a n s that starved females p r o d u c e e g g s in 1-2 d a y s w h i c h is in
a c c o r d a n c e with the opinion of Scott & Baynes (1978). The e x p o n e n t i a l p h a s e b e g i n s on
d a y 3 after the i n c r e a s e in reproduction rates. The p e a k of the p a r a m e t e r e g g s / o v i g e r o u s
female occurs a d a y later than that of the previous p a r a m e t e r . That is due to the fact that
on d a y 3 the p o p u l a t i o n consists, in the main, of well fed adults c a p a b l e of p r o d u c i n g a
high n u m b e r of e g g s in a relatively short time. These will h a t c h quickly, within 15-30 h of
a p p e a r i n g externally (Snell & Carrillo, 1984; Ydfera, 1987), giving rise in one or two days
to a y o u n g p o p u l a t i o n which m a r k s the start of e x p o n e n t i a l growth. Up to that point, the
difference in the rotifer densities of the five groups is small. On d a y 4, the adults p r o d u c e d
the previous d a y will carry more e g g s giving the p e a k a l r e a d y noted, b u t the r e j u v e n a t i o n
of the p o p u l a t i o n m e a n s that the m a x i m u m ratio of e g g s p e r individual c a n n o t occur on
this day.
The increase in population d e p e n d s on the initial production of e g g s a n d is deter-
m i n e d to a large extent b y the quality of the food. The A N O V A shows that the five diets
p r o d u c e different effects in this respect throughout the 15 days of the e x p e r i m e n t a n d the
difference is n o t i c e a b l e from days 3-4, d e m o n s t r a t i n g a clear i m p r o v e m e n t in a popula-
tion fed on T. suecica, whose individuals p r e s e n t far superior size, n u m b e r of eggs,
fecundity a n d growth rates with K values a v e r a g i n g 0.58-0.65. The y e a s t diet is less
effective in all respects. From the A N O V A it can also b e d e m o n s t r a t e d that the p o p u l a -
tion b e h a v e s differently in the exponential p h a s e c o m p a r e d to the exploitation phase. In
the former, the p o p u l a t i o n is m a i n t a i n e d due to 3-day growth cycles closely l i n k e d to e g g
production cycles. The a b o v e m e n t i o n e d l i n k a g e is less e v i d e n t in Y group w h o s e e g g
production cycles are slightly longer. In the latter, however, the cyclical p r o c e s s e s t e n d to
d i s a p p e a r or to diminish due to the progressive rejuvenation c a u s e d b y harvesting, which
brings as a result the growing p r e s e n c e of i m m a t u r e individuals.
It is quite clear that T. suecica stimulates e g g production, a n d its effect is even
g r e a t e r than that o b s e r v e d b y other authors with other a l g a e (Hirayama & N a k a m u r a ,
1976; Yfifera & Pascual, 1984). In the case of Isochrysis galbana a n d yeast, a daily
harvesting of 50 % of the volume p r o d u c e s a minimal n u m b e r of adults, not sufficient to
m a i n t a i n the population size. This l e a d s to a drop in the reproduction rate a n d a fall in the
density of the populations until they reach m i n i m u m levels. The low n u m b e r of adults
produced, especially w h e n fed on yeast, is p r o b a b l y due to l o n g e r e m b r y o n i c develop-
m e n t times a n d lower growth rates. Y6fera (1987) p o i n t e d out the relationship b e t w e e n
embryonic d e v e l o p m e n t times a n d quahty of a l g a e ingested, so that the former is longer
w h e n the diet quality is poorer. This l e a d s to lower g r o w t h rates, e s p e c i a l l y w h e n
populations are s u b m i t t e d to exploitation, b e c a u s e most individuals are h a r v e s t e d before
maturity is reached.
The nutritional quality of the diets u s e d in this e x p e r i m e n t is quite different (Est~vez
180 M. Planas & A. EstSvez

& Planas, 1988). Data on protein content in diets s h o w e d a similar level in all of t h e m
(about 10 % dry weight). Yeast is highly deficient in lipids (1.9 %) a n d in w - 3 H U F A
(traces) while Tetraselmis a n d Isochrysis h a v e 19.9 a n d 31.2 % lipids, with 6.4 a n d 9.3 %
w-3HUFA. T h e i m p r o v e m e n t of rotifer growth rates after the e n r i c h m e n t of diet with fish
oil (rich in w-3HUFA) has b e e n r e p o r t e d b y H i r a y a m a & F u n a m o t o (1983). T h e fatty acid
20:5n-3 is the m a j o r c o m p o n e n t of w - 3 H U F A in b o t h Isochrysis a n d Tetraselmis strains
used. This fatty acid is a b s e n t in yeast. The lipid content w a s similar in rotifers fed on
diets including a l g a e (about 10 % dry weight). H o w e v e r , the 20:5n-3 c o n t e n t w a s twice as
h i g h in rotifers fed on Tetraselmis (0.55 % dry weight). This could b e one of the reasons
w h y the ingestion of Tetraselmis improves g r o w t h rates. Other nutritional r e q u i r e m e n t s
(vit. B12, cystine) for rotifers h a v e b e e n r e p o r t e d b y Scott (1981) a n d H i r a y a m a &
F u n a m o t o (1983). Most a l g a e h a v e the ability to t a k e up vitamine B12 b u t this v i t a m i n e is
a b s e n t in b a k e r ' s yeast. According to H i r a y a m a & Funamoto, this is one of t h e c a u s e s w h y
mass culture of Brachionus with b a k e r ' s y e a s t is unstable. In the opinion of t h e s e authors,
rotifers fed on b a k e r ' s y e a s t in a b s e n c e of vit. B12 could not grow, a n d their e g g s w e r e not
viable. This could also explain w h y growth rates in Y-group s o m e t i m e s d e c r e a s e while
e g g production increases.
Given that the lorica size is genetically d e t e r m i n e d , e n v i r o n m e n t a l conditions can
only modify it to a smaU extent (Snell & Carrfllo, 1984). Accordingly, m e a n differences in
lorica l e n g t h c a u s e d b y diet are small (7.85 %) a n d are in the r a n g e i n d i c a t e d in the
literature (15 % according to Snell & Carrillo, 1984; 13 % according to Yfifera, 1982; 4.5 %
according to F u k u s h o & Iwamoto, 1981). M e a n size can be d i r e c t l y l i n k e d to the food
particle size as was o b s e r v e d b y Scott & Baynes (1978), p r o b a b l y d u e to a n effect on
filtration rates. Yfifera (1982) p o i n t e d out that in starved conditions b o d y g r o w t h is small
a n d longevity short. The s a m e thing must occur w h e n diets of p o o r nutritional v a l u e for
the rotifers are used, such as y e a s t and, to a lesser extent, I. galbana a s c o m p a r e d to
others of b e t t e r quality, n a m e l y T. suecica.
In the opinion of Yfifera & Pascual (1984), an i n c r e a s e in the l e n g t h of adults a n d in
the size of e g g s brings a longer e m b r y o n i c s t a g e and c o n s e q u e n t l y a d e c r e a s e in the
growth rate. This d e c r e a s e did not occur in this e x p e r i m e n t in the case of T. suecica since
the a v e r a g e production of e g g s for female increased.
The results o b t a i n e d with a m i x e d diet, n o t a b l y T. suecica a n d yeast, are of special
i m p o r t a n c e from an economical point of view since they give g r o w t h r a t e s w h i c h are as
g o o d as those shown b y the a l g a e alone, m a k i n g it possible to obtain satisfactory results
with algal densities a p p r e c i a b l y l o w e r than those u s e d in this e x p e r i m e n t . O t h e r authors
h a v e a l r e a d y p o i n t e d out the beneficial effect of m i x e d diets on rotifers u s i n g a l g a e a n d
b a k e r ' s y e a s t (Hirayama & W a t a n a b e , 1973; Yfifera & Pascual, 1980; J a m e s et al., 1983;
J a m e s et al., 1987). This is due to the e n h a n c e m e n t of the overall p r o d u c t i o n of rotifers b y
s u p p l e m e n t i n g the nutritional deficiency of yeast.

Acknowledgements. The writers wish to thank Dr. Jos~ Iglesias, Alison Readman and Encarna de
Miguel for their help in the preparation of this paper. This work was supported by a grant from the
CAICYT.
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