Aquaculture, 69 (1988) 93-104 93

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

The Effect of Broodstock Ration Size on the

Complosition of Rainbow Trout Eggs (Salmo
gairdineri)

D. KNOX’, N.R. BROMAGE’, C.B. COWEY’ and J.R.C. SPRINGATE”

‘Institute of Marine Biochemistry, St. Fittick’s Road, Tarry, Aberdeen AB13RA (Great
Britain)

‘Fish Culture Unit, University of Aston, Birmingham B4 7ET (Great Britain)

(Accepted 18 September 1987)

ABSTRACT

Knox, D., Bromage, N.R., Cowey, C.B. and Springate, J.R.C., 1988. The effect of broodstock
ration size on the composition of rainbow trout eggs (Salmo gairdneri) . Aquaculture, 69: 93- 104.

Female rainbow trout were given half- (0.35%) or full-ration (0.7% body weight per day) of a
commercial trout pellet for a period of 1 year prior to spawning. During development of the eggs
produced by each group of trout, samples of the eyed eggs, yolk-sac and swim-up fry were analysed
for proximate, elemental, amino acid, fatty acid and enzyme composition.
The eggs produced by the half-ration fish were significantly smaller than those of the full-ration
trout. A number of small differences were also found between the composition of both groups of
eggs. However, the data obtained suggested that ample supplies of the materials essential for
successful embryo production were present in eggs from both half- and full-ration fed rainbow
trout.

INTRODIJCTION

In oviparous vertebrates the developing embryo is totally dependent on the

nutrients stored in the yolk for successful development. This nutrient store is
especially important for salmonids where the period of dependence lasts sev-
eral months and the amounts of yolk are correspondingly large. Clearly, vari-
ations in composition of essential materials in the yolk, possibly induced by
modifications in diet or ration of the broodstock, may subsequently lead to
changes in the patterns of survival of the eggs and yolk-sac fry. Under com-
mercial conditions farmers employ a variety of feeding regimes for their brood-
stock, with little consideration being given to the effects of different ration or
nutrient quality on egg production (Springate and Bromage, 1984).
The present experiment was designed to examine the effect of varying brood-

0044-8486/88/$03.50 0 1988 Elsevier Science Publishers B.V.

94

stock ration intake prior to spawning on the biochemical composition of eggs

during development from the eyed stage, through hatching to the swim-up fry
stage.

MATERIALS AND METHODS

Groups of 40 female rainbow trout (Salmo gairdneri) were maintained in 2-

m circular tanks (approximate volume 4 m3) and fed in replicate at either
0.35% (half-ration) or 0.7% (full-ration) body weight day-l on a dry com-
mercial trout pellet ( Ewos-Baker, Scotland: Pellets 7 and 8; proximate anal-
ysis - total protein 47%, carbohydrate 22%, ash 13%, moisture 9%, fat 7%,
fibre 2% ) . Water inflow to each tank was 40 1 min-’ at a constant temperature
of 10°C. The fish, which were of North American origin, but with an
October/November spawning time under local conditions, were 2 years old at
the beginning of the experiment on 13 October. Individual weighing of all the
fish gave a mean initial weight of 0.5815 0.005 kg. Only fish which had spawned
for the first time during the week before the experiment was begun were used.
The fish were individually marked with numbered tags attached by plastic
ties through the dorsal fin. At approximately monthly intervals fish were in-
dividually weighed under phenoxyethanol anaesthesia (1 in 3000)) and also
checked for signs of sexual maturation. As maturity approached, the fish were
examined each week for the presence of ripe or ovulated eggs. The eggs were
removed from each mature fish by gentle pressure on the abdominal wall (i.e.,
stripping) and separately dry-fertilized in a plastic bowl with an aliquot of
sperm from a pool of milt derived from several ripe male fish. Post-stripped
fish weights and length were also recorded. One minute after fertilization the
eggs were gently washed in slowly running water and after water-hardening
for 1 h in the same bowls, the volume of eggs derived from each female was
determined and the mean diameter of the eggs calculated by counting the num-
ber of eggs which could be aligned along a 30-cm long groove in a measuring
board. The number of eggs from each fish (i.e., the fecundity) was then cal-
culated from the volume of eggs and their mean diameter using the method of
Von Bayer (1950 ) . This method provides a constant measure of fecundity which
is not significantly different from fecundity determinations based on total egg
counts (J.R.C. Springate and N.R. Bromage, unpublished observation,
1984) .The fecundities of the fish on full- and half-rations from this experiment
have been reported in an earlier publication (Springate et al., 1985).
After the fecundity determinations were completed, the eggs from six fish
(three from each treatment), randomly selected from the 70-80 batches avail-
able, were transferred to six egg trays in a single hatchery trough (Ewos Baker).
The water flowing through the trough was a dechlorinated ‘town’ supply with
a temperature of 10 L 1 ‘C. Each of the six batches of eggs was maintained in
 95

this incubation system until the fry, which hatched from these eggs, had reached
the swim-up stage.
At three stages of development, i.e., as eyed eggs ( 24 days post-fertilization),
newly hatched yolk-sac fry (35 days post-fertilization), and swim-up fry (58
days post-fertilization), samples were removed from each of the six batches,
counted and weighed after surface water had ben removed with absorbent tis-
sue. Each sample was then analysed in the following assays.

Enzyme lassays

The eggs were homogenised in 10 ~01s. of ice-cold 0.1 M phosphate buffer

pH 7.4 Ccntaining Triton-X-100 (10 g/l) an d centrifuged for 10 min at 12 OOOg.
The clear supernatants obtained were used to assay the following enzymes:
pyruvate kinase, EC 2.7.1.40, PK (Johnston, 1977), lactate dehydrogenase,
EC 1.1.1.27, LDH (Lush et al., 1969)) aspartate aminotransferase, EC 2.6.1.1.,
AAT ( D’Apollonia and Anderson, 1980)) glutathione peroxidase, EC 1.11.1.9.,
POD (B’s11et al., 1985), glucose-6-phosphate dehydrogenase, EC 1.1.1.49, G-
6-PDH (Lijhr and Waller, 1974), isocitrate dehydrogenase, EC 1.1.1.42., ICDH
(Bernt a.nd Bergmeyer, 1974) and superoxide dismutase, EC 1.15.1.1., SOD
(Panchenko et al., 1975). All assays were performed at 20” C.

Vitamin analysis

To measure ascorbic acid content the eggs were homogenised in 10 ~01s. of

ice-cold .trichloroacetic acid (100 g/l) and centrifuged at 12 OOOgfor 5 min.
The supernatant obtained was used to determine ascorbic acid according to
McGown et al. (1982). cr-Tocopherol levels in the eggs and fry were deter-
mined using the method of Cowey et al. (1981) .

Proximate and mineral analysis

The lipid content of the eggs was determined gravimetrically following ex-
traction by the method of Folch et al. (1957). Total protein was determined by
measuring total nitrogen content using the Kjeldahl method, crude protein
content was obtained by multiplying total nitrogen by 6.25. The water, ash and
mineral content of the eggs was determined as described by Cowey et al. (1977).

Fatty aciidand amino acid analysis

Following extraction of the lipid from all batches of eggs a sample of the
chloroform extract obtained was purged with argon and stored at - 80’ C. Fatty
acid anal.ysis of the lipids was carried out after acid-catalysed transmethyla-
tion of the fatty acids (Christie, 1973). The fatty acid methyl esters were re-
96

solved and measured as described by Cowey et al. (1985). The polar and neutral
lipid fractions of eyed eggs were separated and fatty acid analysis performed
according to Cowey et al. (1985). The total amino acid content of the eggs was
carried out after refluxing of a known quantity of the eggs in a large excess of
6 N hydrochloric acid for 16 h (Roach et al., 1967). A sample of the hydrolysate
was taken to dryness under vacuum, then washed with distilled water and dried.
The process was repeated several times to remove the remaining hydrochloric
acid. The acid-free sample was dissolved in 0.01 N HCl to give an approximate
protein content of 0.2 mg crude protein/ml. The amino acids were resolved and
estimated using an amino acid analyser (model JLC-GAH, Jeol (UK) Ltd.,
Grove Park, London ) .

Statistical analyses

The statistical significance of the experimental data obtained at each devel-

opmental stage was established by Student’s t-tests (Fisher, 1950).

RESULTS

In eggs, produced by broodstock rainbow trout fed either half or full food
ration, the wet weight fell slightly in newly hatched fry, but increased at the
swim-up stage. Comparison of the weights showed that the eggs from brood-
stock fish given half-ration were significantly smaller than those from fish on
full-ration (Table 1) .
During development the proximate composition (lipid, protein, ash and
water) of both batches of eggs remained constant until hatching. After hatch-
ing, the water content of all fry increased, the protein and lipid levels fell sharply,
and there were slight decreases in the ash values (Table 1) . Comparisons of
the groups of eggs at the eyed and newly hatched stages showed no significant
differences in water, lipid, protein or ash content. However, at the swim-up fry
stage the protein and lipid concentrations in eggs from fish on full-ration were
significantly greater.
Measurement of the ascorbic acid content of the eggs showed no significant
difference between the two groups at any developmental stage (Table 1) . The
concentrations of vitamin C at the eyed and newly hatched yolk-sac fry were
similar; however, at the swim-up fry stage the concentration had been reduced
by half in each group. Samples of eggs taken at the eyed and newly hatched
stages of both groups were destroyed during a-tocopherol analysis. However,
measurements of tocopherol levels in the two groups of swim-up fry revealed
no significant differences (Table 1) . The measured activities of a number of
enzymes are shown in Table 2. No differences were found between the eggs
 97

TABLE 1

Wet weight, water, ash, lipid, protein, ascorbic acid and cu-tocopherol levels in developing eggs
obtained from broodstock rainbow trout fed half- or full-ration per day

Broodstock Eyed eggs Yolk-sac fry Swim-up fry

ration

Wet weight Half ’ 63.7* k2.76 58.0 *? 2.41 94.6 * 2 3.87

(mg) Full 82.9 k1.22 75.6 ? 1.50 105.1 f 3.38

Water Half * 0.68 kO.01 0.68 + 0.01 0.83* k 0.01

(g/g wet weight) Full 0.66 + 0.003 0.66 * 0.003 0.79 kO.01

Ash Half ’ 12.63 kO.20 13.11 ? 0.58 10.44 + 0.65

(mg/g wet weight) Full 13.32 & 0.36 14.05 * 0.15 11.90 fO.12

Lipid Half ’ 73.97 k2.79 80.09 f 3.85 30.82* + 2.07

(mg/g wet weight) Full 78.35 + 3.23 92.09 + 4.37 42.61* ?I 1.85

Protein Half 2233.40 & 5.34 231.26 & 7.33 122.33* + 6.55
( mg/g wet weight) Full 240.79 k 3.19 244.70 ? 2.57 149.49 & 5.90

Ascorbic acid Half *112.54 + 8.66 120.47 + 10.12 56.92 + 3.72

(pg/g wet weight ) Full 94.30 + 1.84 95.51 ? 1.83 59.73 III3.63
a-Tocopherol Half ’ 56.07 kO.78
(@g/g wet weight) Full _ 71.42 + 5.51

‘Mean&SEM (n=21).
“Mean?SEM (n=3).
*Significantly different from full-ration group (PC 0.05).

from fish on half- and full-rations at the three developmental stages examined.
During development the activities of all enzymes increased in both groups of
the eggs, with the greatest increments being observed in PK, AAT, LDH and
GDH.
Amino acid analysis revealed no significant differences between the groups
at the eyed egg stage. Comparison of the two batches of eggs at the other two
developmental stages examined revealed a number of small, but significant
differences (Table 3 ) . There were no differences in the amino acid concentra-
tions of the eyed eggs and the newly hatched stage. After hatching, the levels
of all the amino acids except glycine fell sharply in both groups of eggs.
Measurement of the mineral concentrations present in the two groups of
eggs showed some small, but significant differences (Table 4). With the ex-
ception of Fe there was very little change in most of the mineral concentrations
during the first stages of development, i.e., eyed and newly hatched stages. In
both groups of eggs the Fe concentrations measured in the newly hatched fry
were approximately one-third of those found in the eyed eggs. After hatching
the 1evel.sof Ca, Na and K increased in both groups of fry while the concentra-
98

TABLE 2

Enzyme activities in developing eggs obtained from broodstock trout fed half- and full-ration per
day

Enzyme Broodstock Eyed egg Yolk-sac fry Swim-up fry

ration

Pyruvate kinase’ Half 29.6 k2.85 228.4 + 40.81 1165.9+ 80.31

Full 33.9 k2.30 248.7 + 49.05 1136.6 + 109.45

Glutamate Half 16.0 i 1.28 50.7-t 8.34 159.6 + 3.99

dehydrogenase’ Full 13.7 kO.24 41.5* 3.01 163.5 t 10.36
Aspartate half 21.69k3.20 98.6? 14.69 544.2 & 17.17
aminotransferase’ Full 22.5 +2.10 94.2& 8.75 606.9 + 63.29
Lactate Half 108.8 + 3.33 421.4k66.61 1686.6 * 71.44
dehydrogenase’ Fuli 115.1 k8.25 434.9? 46.07 1708.7 f 74.49
Glucose-6-phosphate Half 10.2 kO.79 15.5& 2.36 32.2? 1.59
dehydrogenase’ Full 9.2 f0.29 14.7? 2.04 31.7* 1.22
Glutathione Half 11.3 f0.79 12.3? 0.59 15.4? 0.76
peroxidase’ Full 10.8 kO.49 10.5& 1.03 15.5* 1.68
Isocitrate Half 33.5 k2.48 115.1& 24.76 291.2 + 3.64
dehydrogenase’ Full 33.7 k6.03 108.8? 5.81 323.3 + 24.28
Superoxide dismutase* Half nd. 13.3* 4.73 47.32 3.03
Full nd. 11.1* 1.54 48.7+ 0.70

‘Enzyme activity=pmoles substrate used h-i g wet weight-‘.

‘Enzyme activity=units h-i g wet weight-’ (the amount of SOD required to inhibit the rate of
oxidation of substrate by 50% is defined as 1 unit of activity).

tions of Mg, P, Zn, Fe and Mn decreased. The Cu concentrations remained

relatively constant throughout development.
The proportion of ( n - 3 ) fatty acids remained approximately constant for
both ration treatments in the eyed egg and yolk-sac fry stages but increased
relative to other fatty acids in the swim-up fry. This increase was due primarily
to a marked increase in the relative amount of 22:6 (n- 3) with a correspond-
ing decrease in 16:0 and some monoenes (Table 5). The proportion of (n - 6)
fatty acids remained relatively constant at all three development stages with
both rations but there was an increase in the relative amount of 20:4 (n- 6)
in the swim-up fry stage with a corresponding fall in l&2 (n- 6) and l&3
( n - 6 ) . As a result of these changes the ( n - 3 ) / (n - 6) ratio increased from
about 3.5 in eyed eggs and yolk-sac fry to about 4.5 in swim-up fry. A number
of small, but significant differences were also found in the fatty acid compo-
sition of the polar lipids and triacylglycerol fractions isolated from the eyed
eggs of both groups (Table 6).
 99

TABLE 3

Amino acid levels’ in developing eggs obtained from broodstock rainbow trout fed half- or full-
ration per day

Amino aci’d Eyed egg Yolk-sac fry Swim-up fry

Half-ration Full-ration Half-ration Full-ration Half-ration Full-ration

LYS 22.17 f 0.68* 22.76 f 1.92 19.97* f0.31 22.08 kO.63 10.27 + 0.44 11.56 + 0.50
His 6.96 & 0.39 7.44 & 1.00 6.63 & 0.22 7.15 kO.53 3.75 +0.26 3.71? 0.26
Arg 16.13 & 0.32 16.64 + 0.76 14.60* + 0.28 16.09 f 0.28 7.88 kO.33 8.55 ?z0.19
Asp 22.15i0.32 23.06 f 0.42 21.13* + 0.56 23.25 + 0.46 12.04 & 0.32 13.53 f 0.46
Thr 11.79+0.18 12.16kO.22 11.13 f0.34 12.31kO.33 5.90* ? 0.18 6.85? 0.28
Ser 14.32 + 0.20 14.62 + 0.39 13.28* + 0.43 15.07 + 0.48 6.15 kO.13 7.32 ?z0.41
Glu 29.59 + 0.53 31.01+ 1.12 28.36 +0.69 31.51+ 1.05 17.75* * 0.40 19.70 * 0.51
GIY 6.65 + 0.05 6.71i0.11 6.78*+0.22 7.52+0.11 6.88 kO.20 7.05 ? 0.02
Ala 20.83 ? 0.30 21.18kO.31 18.59*?0.59 21.52f0.67 7.54* + 0.25 9.36 f 0.60
CYS 3.73 & 0.09 3.07? 0.47 2.88 ? 0.11 3.75 f0.33 1.46 +0.13 1.50 * 0.29
Val 14.49 * 0.31 16.06? 0.90 17.31 ? 0.73 18.79 + 0.52 6.90* + 0.22 8.76 + 0.57
Met 7.71 f0.09 7.76 +0.09 7.20 ? 1.08 8.02 k 0.13 3.65 +0.28 4.37 f 0.27
Ileu 14.01 kO.23 14.44 i 0.32 12.79* i 0.43 14.29 f 0.19 5.52 50.28 6.72 +0.51
Leu 23.62 f 0.50 24.36 ? 0.37 22.56 f 0.70 24.33 + 0.22 9.76 kO.39 11.81?0.75
Tyr 10.36&0.11 10.67f0.12 9.70 f0.49 11.01?0.17 4.62 +O.lO 5.65 ? 0.36
Phe 12.95 If:0.15 13.19+0.10 12.50 +0.66 13.62f0.29 5.79 f0.20 6.72 ? 0.29
Pro 12.47 f0.19 13.20+0.18 11.23 kO.34 12.18kO.36 5.15*+0.12 6.33 f 0.30

‘mg Aminmoacid/g wet weight.

‘MeanfSEM (n=3).
*Significantly different from the full-ration group (PC 0.05).

TABLE 4

Mineral concentrations’ in developing eggs obtained from broodstock rainbow trout fed half- or
full-ration per day

Mineral Eyed egg Yolk-sac fry Swim-up fry

Half-ration Full-ration Half-ration Full-ration Half-ration Full-ration

Ca (mg) 0.59 f 0.08’ 0.54 + 0.08 0.57 f 0.08 0.55 + 0.08 0.69 kO.08 0.76 k 0.03
Mg (mg) 0.53 * 0.01 0.46 + 0.03 0.57 f 0.02 0.53 f 0.02 0.26 kO.01 0.30 * 0.02
P (mg) 2.76kO.14 2.78kO.13 3.62 kO.04 3.77kO.11 2.30’ ? 0.13 2.78 + 0.06
Na (mg) 0.21 kO.003 0.20 kO.01 0.29*? 0.01 0.23 f 0.01 0.82 20.05 0.69 i 0.02
K (mg) 1.58kO.01 1.63f0.04 1.81*?0.05 1.98kO.02 2.44 fO.O1 2.34? 0.08
Gu (pg) 2.27 kO.16 2.36 f 0.25 1.63 ? 0.15 2.07 f 0.23 1.40 kO.13 1.98kO.24
Zn (pg) 26.21 f 1.56 23.19 k 0.39 28.91* ?z1.19 24.62? 0.91 18.25 f 0.99 17.71& 1.40
Fe (pg) 60.12 + 8.68 52.35 + 3.68 18.42 + 1.39 15.36& 0.61 13.34 f 1.88 12.13? 1.54
Mn (pg) 0.77 f 0.02 0.99 i 0.26 0.65 + 0.02 0.74 + 0.04 0.47 kO.02 0.50 i 0.02

‘Results expressed mg (or pg) per g wet weight as indicated.

‘Mean?SEM (n=3).
*Significantly different from the full-ration group (P < 0.05 ) .
100

TABLE 5

Fatty acids’ of biochemical interest in the total lipid of developing eggs obtained from rainbow trout
broodstock fed half- or full-ration per day

Fatty acids Eyed egg Yolk-sac fry Swim-up fry

Half-ration Full-ration Half-ration Full-ration Half-ration Full-ration

140 1.61 50.12’ 1.53&0.10 1.58 kO.01 1.56i0.19 1.07 TO.08 1.08iO.08
16:0 16.24’ ? 0.44 14.13 f 0.62 16.05 & 0.59 14.18? 1.00 13.40* f 0.40 11.92 Z!I0.27
16:l (n-7) 5.85 kO.42 6.55f0.67 5.57 kO.17 6.67 k 0.68 3.78 ? 0.21 4.80 kO.39
l&O 5.27 kO.09 5.31 ItrO.26 5.41 kO.12 5.16kO.12 4.81 f 0.06 4.94 +0.18
18:l (n-9) 21.47 20.31 22.91+0.71 21.32 ? 0.64 22.971t1.61 16.77*&0.19 20.58f0.98
18:l (n-7) 4.57 kO.16 5.33kO.24 4.46*?0.11 5.24 + 0.16 3.35*?0.09 4.18+0.21
18:2 (n-6) 4.46 kO.04 3.80f0.14 4.20 kO.07 3.89 + 0.29 3.32 +0.08 3.38f0.18
18~3 (n-6) 0.38 kO.02 0.34f0.01 0.19 kO.05 0.10+0.003 0.16 kO.02 0.15f0.01
l&3 (n-3) 0.28 f0.02 0.19 ? 0.03 0.36 + 0.01 0.34? 0.01 0.28 & 0.02 0.29 ? 0.01
l&4 (n-3) 0.28 kO.02 0.19 + 0.03 0.27 ? 0.01 0.17 + 0.04 0.25 kO.01 0.18f0.04
2O:l (n-9) 0.29 20.01 0.23 + 0.02 0.29 t 0.003 0.22 * 0.02 0.22 i 0.01 0.20 * 0.003
2O:l (n-7) 2.87* f 0.03 3.71 f0.17 2.96 +0.02 3.57 ? 0.26 2.44*?0.06 3.46f0.14
20:2 (n-6) 0.76 f0.03 0.93 ? 0.06 0.76 f 0.03 0.95kO.15 0.70* f 0.03 0.99 AZ0.10
20~3 (n-6) 1.27 *0.19 l.ll+O.lO 1.24 i0.16 1.12io.10 1.22 io.21 1.17io.10
20~4 (n-6) 1.91 kO.14 1.46 ? 0.34 1.93 ?I 0.08 1.54 f 0.30 2.59 f 0.08 1.95 IL0.33
20:4 (n-3) 0.31 kO.02 0.42 + 0.09 0.30 ? 0.01 0.41* 0.10 0.31 f 0.01 0.42 If-0.08
20:5 (n-3) 4.31**0.17 3.27t 0.30 4.16 ? 0.12 3.23 IL0.28 4.55**0.15 3.35t0.17
22:l (n-11) 0.30 fO.O1 0.30 + 0.06 0.23 2 0.01 0.27 + 0.003 0.27 & 0.05 0.35 IL0.10
22:l (n-9) 0.15 fO.O1 0.19 kO.02 0.12 & 0.01 0.17~0.003 0.19 fO.O1 0.20~0.01
225 (n-3) 1.59 20.01 1.40?0.13 1.53 zLo.14 1.39 kO.25 1.79 +0.08 1.5550.16
22:6 (n-3) 21.82 fO.44 21.94f 1.09 21.35 ?I 1.41 22.07 + 2.83 31.64 TO.40 28.50 & 1.29

‘Values are % by weight of total fatty acids.

‘Mean?SEM (n=3).
*Significantly different from the full ration group P < 0.05.

DISCUSSION

The differences observed in the egg weights of half- and full-ration fed
broodstock trout confirm the earlier findings of Springate and Bromage (1984)
and Springate et al. (1985). Thus the eggs/fry obtained from broodstock given
a full daily ration were significantly heavier than those obtained from trout on
half-ration.
The changes in the proximate composition of both groups of eggs during
development were very similar to those reported by Ando (1962 ) and Suzuki
and Suyama (1980). No significant differences were found between the two
egg groups at the eyed egg or yolk-sac fry stage. However, in the swim-up fry
examined, both protein and lipid levels were significantly greater in those fry
obtained from broodstock fish given the full daily ration. This could perhaps
indicate that the period between yolk-sac absorption and the onset of feeding
may be a critical phase for half-ration fry since endogenous energy sources
could be limited.
 101

TABLE 6

Fatty acids’ of biochemical interest in polar lipids and triacylglycerols of eyed eggs obtained from
rainbow trout broodstock fed half- or full-ration per day

Fatty acid Polar lipids Triacylglycerols

Half-ration Full-ration Half-ration Full-ration

14:o 0.92 k 0.012 0.87kO.11 1.83 kO.08 1.75 * 0.22

16:0 17.38 IL0.56 16.28? 0.34 16.54 + 0.57 15.03 + 0.73
16:l (n-7) 2.34 +0.17 2.79? 0.38 7.78 f0.47 9.88? 0.81
18:0 8.40* f 0.30 9.38? 0.14 3.79 kO.56 3.70 + 0.08
18:l (n-9) 10.33 + 0.57 11.55kO.33 27.76*? 1.37 33.82 + 1.47
18:l (n-7) 4.07* IL0.09 4.83 ?I 0.06 4.82* +0.17 6.25 f 0.23
1812 (n-6) 2.39 kO.22 3.02 k 0.71 5.28 kO.44 4.98 k 0.33
18:3 (n-6) 0.28 kO.05 0.29 k 0.04
18:3 (n-3:1 tr tr 0.45 20.05 0.42 k 0.01
18:4 (n-3:1 0.35 kO.03 0.25 + 0.01
2O:l (n-9’, tr tr 0.30 f0.02 0.30 * 0.01
2O:l (n-7 I 4.25* f 0.04 5.35kO.15 2.74 +0.46 3.58kO.15
20:2 (n-6.1 0.92 f0.18 0.98 +O.ll 0.39 +0.03 0.58kO.13
20:3 (n-61 1.47 kO.03 1.46 +0.12 0.81 f0.17 0.66 k 0.09
20:4 (n-61 3.10 k0.32 2.46 ? 0.76 1.27 kO.23 0.81 kO.ll
20:4 (n-3) 0.33 kO.03 0.40 k 0.08
20x5 (n-3 I 5.02 kO.14 3.68 k 0.34 3.03* L 0.33 1.98 k 0.09
225 (n-3 I 1.72 k0.26 1.62 k 0.06 1.03 +0.11 0.78kO.11
22:6 (n-3 1 30.52 k 1.89 29.62 k 1.13 12.89 ? 1.46 10.80 + 1.46

‘Values are % by weight of total fatty acids.

*Mean&SEZM (n=3).
*Significantly different from the full-ration group (P<O.O5)
tr = < 0.1% by weight of total fatty acids.

The observed changes in the mineral composition of both egg groups during
development are similar to those described by Ogino and Yasuda (1962)) i.e.,
post-hatching levels of Ca, Na and K increase while the concentrations of Mg,
P, Cu, Zn, Fe and Mn decrease. Although a number of differences were found
between the eggs from fish on half- and full-rations the indications are that
the size of the broodstock ration had little effect upon the mineral composition
of the eggs produced.
The enzyme activities measured in both groups of eggs showed no differences
between the half- and full-ration groups. Thus, glycolytic capacity (PK and
LDH ) , a.mino nitrogen handling (AAT and GDH ) , reducing equivalent pro-
duction ( ICDH and G-6-PDH) and cell defences against oxygen toxicity (SOD
and POD) were very similar in both batches of eggs. Ascorbic acid and LY-
tocopherol are also involved in the protection of cell lipids against peroxidation
and, in common with the measured enzyme levels, broodstock ration size had
no effect upon the levels of these vitamins in the eggs produced. The decreases
102

in ascorbic acid levels during development were also similar to those found in
Atlantic salmon eggs (Cowey et al., 1985).
Studies of the nutritional requirements of rainbow trout indicate that n - 3
fatty acids are essential for the maintenance of good health and rapid growth
(Watanabe et al., 1974). Comparison of the total II - 3 fatty acid levels in the
lipid isolated from the developmental stages of both groups of eggs showed no
significant difference between the eggs from fish on half- and full-rations.
Highly unsaturated long chain fatty acids ( > 20:5) are generally seen as es-
sential in the maintenance of membrane fluidity. In the polar lipid fractions
of eggs from both groups the long chain fatty acids accounted for more than
30% of the total present, a value similar to that found in eggs of Atlantic salmon
(Cowey et al., 1985). Differential rates of oxidation of fatty acids appear to
occur during development; saturated acids and monoenes being oxidised more
rapidly than the highly unsaturated fatty acids 20:4 ( n- 6) and 22:6 ( n - 3 ) .
This is presumably the means by which the relative proportion of these acids
is increased in swim-up fry. Thus although a number of minor differences were
found in the fatty acid composition of half- and full-ration eggs, overall one
must conclude that broodstock ration size produces no major changes in the
fatty acid content of the eggs.
The amino acid composition of the eggs from the two groups of fish shows a
similar picture to the fatty acids, i.e., the essential amino acids for health and
growth in rainbow trout are present in similar concentrations in both half- and
full-ration eggs. The changes in amino acid levels during egg development are
also similar to those described by Suyama (1958). Thus broodstock ration size
had no major effect upon the amino acid composition of the eggs produced.
One of the more significant effects of feeding half-ration was a reduction in
the size of the eyed eggs produced. This finding confirms earlier studies of
Springate and Bromage (1984) and Springate et al. (1985). These authors
have also shown that the quality or survival of the eggs and fry derived from
small eggs are similar to those achieved from larger eggs (Springate and Brom-
age, 1984; Springate et al., 1985). If egg quality is determined by the compo-
sition of eggs then the absence of any differences in either the quality of the
eggs and fry or the chemical and biochemical composition of the eggs from the
two ration groups would tend to support the proposition of Phillips and Dumas
(1959) that ample supplies of material for viable embryo production are pres-
ent even in the smallest of eggs. From the commercial standpoint it is impor-
tant to establish that modifications of ration employed either to optimise egg
production from broodstock (Springate and Bromage, 1984; Springate et al.,
1985) or to reduce farm effluent can be used without risk to the quality or
composition of the eggs produced.

ACKNOWLEDGEMENT

This study was supported in part by a NERC Research Grant to N. Bromage.

 103

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