Download Effect of feeding method and protein source on Sparus

Aquaculture 224 (2003) 89 – 103
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Effect of feeding method and protein source
on Sparus aurata feeding patterns
M.J. Sánchez-Muros a,*, V. Corchete b, M.D. Suárez a, G. Cardenete c,
E. Gómez-Milán a, M. de la Higuera c
a
Department of Applied Biology, University of Almerı́a, 04120 Almerı́a, Spain
Department of Applied Physics, University of Almerı́a, 04120 Almerı́a, Spain
c
Department of Animal Biology and Ecology, University of Granada, 18071 Granada, Spain
b
Received 5 February 2002; received in revised form 14 February 2003; accepted 18 February 2003
Abstract
The influence of dietary protein source (fishmeal, soy-protein concentrate and soy-protein
concentrate supplemented with methionine) on voluntary feed intake, daily feeding rhythm and
nutritive utilisation of diet was studied in the gilthead sea bream (Sparus aurata) fed by hand or
demand feeding. Fish weighing 21 g were maintained indoors under natural conditions of
temperature and photoperiod (transparent ceiling) and allowed ad libitum or self-feeding of
experimental diets for 26 days, with three replicates per treatment. In the first experiment, the
influence of hand or demand feeding on growth rate and feed utilisation of a fishmeal-based control
diet was studied. In a second trial, involving different protein sources, fish were maintained under the
experimental conditions for 6 days after 20 days training. The general composition of the
experimental diets was: 45% protein, 14% lipids and 20% carbohydrates. Results showed that: (1)
gilthead sea bream preferentially feed in the afternoon and evening; (2) demand feeding improved
both food conversion and protein efficiency; (3) the protein source appeared to induce changes in the
timing of feeding; and (4) supplements of methionine advanced the time of feeding and lengthened
ingestion phases.
D 2003 Elsevier Science B.V. All rights reserved.
Keywords: Feeding behaviour; Feed intake; Sparus aurata; Sea bream; Soy-protein; Methionine
* Corresponding author. Fax: +34-950-015476.
E-mail address: [email protected] (M.J. Sánchez-Muros).
0044-8486/03/$ - see front matter D 2003 Elsevier Science B.V. All rights reserved.
doi:10.1016/S0044-8486(03)00211-4
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1. Introduction
Maximum diet performance involves two important advantages: reduction of feeding
costs and decreased waste. The study of feeding behaviour in several fish species has
revealed that the adjustment of feeding times to match natural rhythm improves nutritional
efficiency, feeding frequency, food conversion efficiency and can even vary the utilisation
of certain nutrients (Bolliet et al., 2001).
Timing of feeding appears to influence the growth of fish. In Heterobranchus longifilis,
growth improved when fish fed at night rather than during the day (Kerdchuen and
Legendre, 1991). In Heteropneustes fossilis, growth was enhanced when feed was
supplied in the scotophase rather than at dawn (Sundaranaj et al., 1982). Thus, the same
food ingested at different times of the day is absorbed with differing efficiencies (Madrid,
1994). Likewise, voluntary daily feeding peaks have been recorded in species such as
European sea bass Dicentrarchus labrax (Anthouard et al., 1993; Sánchez-Vázquez et al.,
1995), rainbow trout Onchorhynchus mykiss (Alanärä, 1996), siluridae (Boujard and
Luquet, 1996), turbot Psetta maxima (Burel et al., 1997), and gilthead sea bream Sparus
aurata (Anthouard et al., 1996).
An ability to regulate the intake of macronutrients has been demonstrated in some
species, such as sea bass (Rubio et al., 2001), rainbow trout (Sánchez-Vázquez et al., 1999)
or goldfish Carassius auratus, the latter even having an ingestion peak for each macronutrient (Sánchez-Vázquez et al., 1998a). Hence, the dietary amino acid profile and
availability (as free or proteic amino acids) could affect feeding patterns.
Protein is a basic component of fish diets, both in terms of quantity and quality, protein
requirements being higher than those of other animals (Cowey, 1975). Fishmeal provides
an adequate balance of amino acids, but its increasing demand and price, as well as
uncertain supply, make it necessary to find alternative protein sources. Furthermore,
problems associated with animal proteins make vegetable proteins the most promising
candidates. The high quality of fishmeal proteins makes substitution difficult; however,
partial substitutions are being made (Shiau et al., 1987; Reigh and Ellis, 1992) and
fishmeal dietary levels could be reduced even further by adequate supplementation with
essential amino acids, providing that the availability of the supplementary amino acids
coincides with that of protein amino acids at the sites of protein synthesis. An adequate
postprandial pattern of amino acids is not only necessary to ensure protein synthesis and
growth, but might also determine feed acceptability. In the case of using soy-protein to
feed gilthead sea bream, the maximum substitution to maintain similar growth to that with
fishmeal is around 20% (Robaina, 1995).
Vegetable proteins generally have an inadequate balance of amino acids, being deficient
in some essential amino acids (Murai, 1992). Soy-protein supplementation with methionine allows significant replacement (50%) of fishmeal (Viola et al., 1982) but not complete
substitution. It has been suggested that the inefficiency of supplementation is due to the
faster uptake and subsequent catabolism of the supplemented amino acids with respect to
protein ones (Cowey and Walton, 1988). In fact, the utilisation of supplemented amino
acids for protein synthesis and growth is improved when they are coated, as shown in carp
(De la Higuera et al., 1998) and gilthead sea bream (Sierra, 1995). Another possibility of
improving dietary free amino acids for growth would be by increasing feeding frequency,
M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
91
with the aim of making all amino acids available at the same time and at the sites of protein
synthesis. However, since sea bass can discriminate between diets containing different
amounts of methionine (Hidalgo et al., 1988), it may be possible for gilthead sea bream to
adapt its feeding behaviour when facing either a methionine-deficient or a free-methionine-supplemented diet.
The aim of the present study is to determine the effect of the feeding method and
protein source on S. aurata feeding patterns. In this sense, we seek to establish (1) whether
demand-feeding improves the notional utilisation of diet for growth; (2) whether gilthead
sea bream displays preferential feeding times throughout the day when fed on demand
with unrestricted access to feed; and (3) whether soy-protein-based diets, supplemented or
not with free methionine, affect voluntary-feeding patterns.
2. Material and methodology
2.1. Animal-rearing conditions
Gilthead sea bream of an initial weight of 21.0 F 3.5 g, obtained from a local fish farm
(Carmar, Carboneras, Almerı́a, Spain), were transported to the University of Almerı́a and
placed in 250-l tanks with sea water at a flow of 10.4 l/h. The fish were kept under ambient
temperature (15.5 F 0.6 jC) and photoperiod. Prior to the experiments, the fish were
divided into two groups, one fed by hand and the other by demand using Aquarium MA32 self-feeders (Procam Ingenieria), as described by Sánchez-Vázquez et al. (1994) that
delivered approximately 9– 11 pellets (about 0.3 g feed/pellet) each time a fish activates
the rod located 3 cm below the water surface. Demand-fed fish were given a 20-day
training period. The experiments started when the fish appeared to have learned to feed
themselves on demand, following the criteria described below.
Feed-intake demands were recorded daily from the beginning of the training period.
Feeders were checked every day and the remaining food weighed. An approximate weight
of uneaten pellets at the bottom of the tanks was used to calculate the amount of food
ingested. The recording of early-day self-feeding was sporadic. Over the next few days,
the fish learned self-feeding, reacting to the food after hitting the bar. After 15 – 19 days,
the request for food approached food intake, and the self-feeding became more regular,
with feed demands concentrating within some hours of the day. The experiment started
when this behaviour remained constant for at least 3 days.
2.2. Diet
The mean general composition of the experimental diets was: 45% protein, 14% lipids
and 20% carbohydrates. Fishmeal (diet D-1), soy-protein concentrate (diet D-2) and soyprotein concentrate supplemented with free methionine (diet D-3) were the protein sources
used as the only source of protein. The ingredients (Table 1) were blended with sodium
alginate and then thoroughly stirred with distilled water to obtain a moist, homogeneous
mixture. Pellets were made as described by Sánchez-Muros et al. (1998). Gross-energy
content of the experimental diets was calculated using metabolizable-energy values of
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M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
Table 1
Ingredients and proximate composition (% dry weight) of experimental diets
Control (D-1)
Met-deficient (D-2)
Met-supplemented (D-3)
60.4
–
–
7.28
11.90
6.65
2
3
2
0.5
1
5.43
–
55.13
–
13.69
6.78
5.79
2
3
2
0.5
1
10.11
–
54.1
0.77
13.69
6.78
5.79
2
3
2
0.5
1
10.11
Proximate composition (% dry matter)
Crude protein
47.71
Fat
13.21
Ash
10.23
Gross energy (MJ/kg)
22
47.30
13.42
11.45
22
47.23
13.41
11.02
22
Ingredient (g/100 g)
Fish meal
Soy meal
Methionine
Fish oil
Corn starch
Dextrin
Vitamin mixturea
Mineral mixturea
Sodium alginate
Cr2O3
Betaine
Micronized cellulose
a
Vitamin and mineral mixture according to Sierra (1995).
19.6, 39.5 and 17.2 kJ g 1 for protein, lipids and carbohydrates, respectively (Brett and
Groves, 1979). Crude protein, total lipids and moisture were analysed by standard methods
(AOAC, 1984). Methionine supplement for diet D-3 was calculated after determining the
amino acid content of the protein sources. Free methionine was added to soy-protein diet
to reach fishmeal-diet levels.
2.3. Experiment 1
Experiment 1 lasted from the 25th of February (sunrise 07:54 h; sunset 19:04 h) to the
21st of March (sunrise 07:10 h; sunset 19:30 h). The performance with hand feeding was
compared to that of demand feeding for 26 days. Groups of 15 fish were used: 3 groups
were hand-fed twice daily (at 10:00 h and 16:00 h) to apparent satiation and the other 3
were self-fed. All groups were fed a fishmeal-based diet (designated ‘‘Control’’ in Table
1). After about 30 min, non-ingested food was removed from hand-feeding tanks, those
having self-feeders were checked frequently. Uneaten food was quantified to calculate the
amount ingested.
2.4. Experiment 2
Fish trained for demand-feeding were divided into 9 groups (three replicates per
treatment) of 15 fish each. Each replicate per treatment was fed the corresponding test diet
for 6 days. Experimental diets had similar macronutrient compositions but varied in
protein source and methionine content (Table 1).
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93
2.5. Spectral data analysis
Feed-demand results were subjected to spectral analysis based on the fast Fourier
transform. Spectral analysis was applied to the recordings of daily food delivered to each
tank, in order to identify any feeding rhythms of voluntary intake. Thus, the Fourier
spectrum H( f ) of the recording sample denoted by h(t) was obtained (Brigham, 1988)
Z l
Hðf Þ ¼
hðtÞej2pft dt
l
pffiffiffiffiffiffiffi
where f is frequency, t is time and j is the imaginary unit ðj ¼ 1Þ. In general, H( f ) is a
complex quantity called Fourier transform of h(t). H( f ) can also be expressed as
Hð f Þ ¼ Rð f Þ þ jIð f Þ ¼ AHðf ÞAe jUðf Þ
where R( f ) is the real part of H( f ) and I( f ) is the imaginary part. The amplitude spectrum
of h(t) is denoted by AH( f )A and the phase spectrum is denoted by U( f ). These are
defined as
pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Iðf Þ
AHðf ÞA ¼ R2 ðf Þ þ I 2 ðf Þ
Uðf Þ ¼ tan1
:
Rðf Þ
When studying the amplitude spectrum of h(t), it is possible to identify the principal
harmonic components of the sample because they always appear with major amplitude
(Bath, 1982). Frequencies corresponding to principal harmonic components are the
frequencies of the rhythms present in the sample (Aschoff, 1992). These rhythms are
identified visually and their period T (T = 1/f ) can be read directly from the amplitude
spectrum (see Fig. 4). By reading the period value at the highest peak points of the
amplitude spectrum, the phases of maximum feeding rhythms were derived for each group
studied. The spreading of these peaks reveals the intake phase or time during which the
maximum intake occurs (Papoulis, 1962; Corchete et al., 1989). The phase spectrum is
considered to determine the timing in which feed demands began to increase, because this
time is the phase delay of the principal harmonic component considered (Papoulis, 1962;
Bath, 1982). The harmonic component with major amplitude corresponds to the daily
rhythm. This harmonic component is easily identified in the amplitude spectrum as
described above. Thus, we can read the phase value for this period in the phase spectrum.
In this way, the phase delay, or timing in which demands began to increase, was derived.
As a result, spectral analysis can be used to identify and quantify any rhythm in feed
demand. Today, the spectral analysis is considered a standard tool for the analysis of time
series in several scientific fields. Particularly, in biomedical sciences, spectral analysis
proves to be a powerful analysis technique to reveal information which is hidden in the
original data and which is extremely difficult to discover with other analysis techniques
(Childers, 1989; Aschoff, 1992). This information hidden in the time domain may possibly
be easier to discover in the frequency domain (Corchete et al., 1989). This powerful
technique has been applied by Corchete et al. (1995) in another field of science, as
geophysics, with successful results.
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Fig. 1. Daily feeding demand of gilthead sea bream fed a fishmeal-based feed recorded in 26 days (experiment 1).
Each graph corresponds to the record for 1 day and represents the average demand of three tanks. All graphs are
normalized to have the maximum of 1. Shadow zones show the hours of darkness.
M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
95
Fig. 2. Average of the 26 records plotted in Fig. 1 (experiment 1). Y-axis units are number of demands in 10 min.
Shadow zones show the hours of darkness. Horizontal arrow shows the intake phase.
2.6. Biological indices and statistical data analysis
The biological indices determined were: feed conversion ratio (FCR), defined as dry
feed intake/biomass gain; and protein efficiency ratio (PER), defined as wet weight gain/
protein intake. Student’s t-test was used to compare the averages of the biological indices
determined.
3. Results
Over the first experiment, fish exhibited a certain feeding pattern throughout the
day; that is, feeding demands increased during the afternoon, starting at about 17:00 h
and ending at about 23:00 h, with a maximum of demands between 19:00 and 20.00 h
Table 2
Growth (W0 and Wf means initial and final weight, respectively), feed intake, feed efficiency ratio (FER) and
protein efficiency ratio (PER) of gilthead sea bream fed using different feeding systems (mean F standard
deviation)
Wf W0 (g)
(Wf W0)/fish (g)
Feed intake (g)
Feed intake/fish/day (g)
FCR (feed intake/weight gain)
PER (weight gain/protein intake)
Hand
Demand
19.95 F 2.10
1.33 F 0.15
52.50 F 4.32
0.14 F 0.01
2.63 F 0.78a
0.80 F 0.29
27.45 F 2.50
1.83 F 0.17
45.00 F 5.06
0.12 F 0.02
1.64 F 0.03
1.28 F 0.54
FCR: feed conversion ratio = dry feed intake/wet weight gain.
PER: protein efficiency ratio = wet weight gain/protein intake.
a
Significant difference between both columns for p < 0.05 (Student’s t-test analysis).
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M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
Fig. 3. Daily feeding rhythms of gilthead sea bream fed on diets of: (a) soy protein supplemented in methionine;
(b) soy protein deficient in methionine; and (c) fishmeal as control diet (experiment 2). On the left side, each
graph corresponds to the record for 1 day and represent the average demand of three tanks for each treatment (all
graphs are normalized to have the maximum of 1). Shadow zones show the hours of darkness. On the right side,
average of the six records plotted in the left side for each treatment. Y-axis units are number of demands in 10 min.
Shadow zones show the hours of darkness. Horizontal arrows show the intake phases.
M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
97
(see Figs. 1 and 2). During the experiment, maximum intake was registered between March
3 and March 5, a period which coincided with a full moon. However, due to the duration of
the experiment, it cannot be statistically determined whether the moon influenced the
amount of feed ingested by the fish. On March 19, coinciding with new moon, there was
also an increase of food consumption associated with a rise in temperature of 1.5 jC. Fish
fed on demand had a significantly lower FCR than those fed by hand, although the
improvement in protein utilisation (PER) did not significantly differ (Table 2).
In the second experiment, the data recorded showed that the general feeding pattern was
maintained, the maximum intake being concentrated in the afternoon/evening, irrespective
of the diet composition (see Fig. 3). However, the soy-protein-based diet supplemented
with free methionine appeared to lengthen the intake phase until 2:30 h (right part in Fig.
3). In this experiment, the intake phases for each diet were: 7:05 h for the control diet (D1), 7:05 h for soy-protein-methionine-deficient diet (D-2) and 9:30 h for the soy-protein
diet supplemented with methionine (D-3). The spreads of peaks in the amplitude spectrum
were considered to establish the intake phases (see Fig. 4 as example of spectral analysis).
Feed demands for the control diet (D-1) began to increase at 17:10 h (the spectrum phase
was considered to determine this time) and the timing for maximum demand-feeding
Fig. 4. An example of spectral analysis applied in this study: (a) Sample record with daily feeding demand for 6
days of gilthead sea bream fed on diets of soy protein deficient in methionine, as showed in Fig. 3b (left side). (b)
Normalized amplitude spectrum of this sample data. (c) A zoom view of the sample record spectrum shown in
panel b. Both spectrum graphs are normalized to have the maximum amplitude of 1.
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M.J. Sánchez-Muros et al. / Aquaculture 224 (2003) 89–103
activity was between 19:00 and 20:00 h (see Fig. 3). For diet D-2 (methionine-deficient),
feed demands began to increase at 17:10 h and the maximum occurred between 19:00 and
20:00 h. For the diet supplemented with methionine (D-3), demands started at 14:20 h,
reaching a maximum between 17:30 and 19:30 h. In the second experiment only, the
spectral analysis revealed a second rhythm with a period of 12 h for each experimental diet
(see Fig. 4). This secondary rhythm had a frequency of two peaks per day, the second peak
coinciding approximately with the peak of the main rhythm.
4. Discussion
Three behaviour patterns were identified in the experimental sea bream: daytime,
nighttime and evening (Aschoff, 1992). A dual behaviour has been described for the
European sea bass (Sánchez-Vázquez et al., 1995, 1998b), and a seasonal change in the
turbot (Müller, 1978); although, in larvae and juveniles of this latter species, daytime
behaviour was described by Burel et al. (1997) and Huse (1994). According to the results
of the present study, gilthead sea bream showed greater feed demand in the evening,
although a certain amount of demand remained throughout the day with a slight surge at
dawn. These results, showing two daily peaks, partly coincide with those of Anthouard et
al. (1996) for this species, who found another peak at midday, although this third peak may
have resulted from a higher experimental temperature (18 – 24 jC) than that of the present
study (14.5 –16.5 jC). In fact, during the second experiment, a secondary rhythm with a
period of 12 h at a temperature 1.5 jC higher was found, while in the first experiment that
secondary peak was not detected, probably because the temperature was maintained
between 14 and 15 jC. One noticeable feature was the considerable increase in demandfeeding in all the tanks during the 2 days of the full moon, although we cannot confirm
these results, since the experiment only lasted 1 month. Lunar cycles have previously been
shown to affect juvenile European sea bass growth rhythms (Planes, 1993). Growth in
length, body weight and food intake have also been reported to be influenced by lunar or
semi-lunar rhythms in several species, such as coho salmon Oncorrhynchus kisutch,
rainbow trout O. mykiss, and Arctic char Salvelinus alpinus (for a review, see Madrid et al.,
2001). However, no reference concerning the influence of lunar cycles for gilthead sea
bream or other Mediterranean species is available, nor for the relationship between
moonlight and feeding behaviour. Therefore, we cannot state whether this effect is a
lunar rhythm or an effect due to the greater clarity of the full-moon nights. This effect
could not be tested for full-moon nights with a cloudy sky. Juvenile silver barb (Haroon
and Pittman, 1997) reportedly feed more actively on full-moon nights. Since gilthead sea
bream were housed in tanks in the present study, the increased intake appears to be due
rather to moonlight than to the tides caused by the moon’s gravitational pull. At the end of
the experiment, demand feeding increased further, coinciding with the new moon,
although this may have been due to a water temperature increase of approximately 1.5
jC during the last 4 days of the assay. The effect of temperature on fish food intake is well
known (Kestemont and Baras, 2001). Nonetheless, a combined effect of the new moon
and temperature cannot be dismissed although no reference new moon effects on food
intake is available in the literature.
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99
Nutritional utilisation of the diet appears to be influenced by the time of feeding, since
the parameters of nutritional efficiency improved when the fish were allowed to feed at
will. In this way, they regulate their intake to meet energy requirements (Kaushik and
Médale, 1994) according to their natural feeding rhythms (Boujard and Leatherland,
1992a).
The FCR and PER results showed that self-fed fish performed significantly better than
animals hand fed at set hours. This could be due to the feeding methods or feeding time. In
fact, it has been demonstrated that the same diet offered at different times of the day is
assimilated differently (Madrid, 1994). Different growth rates have been recorded in
H. longifilis (Kerdchuen and Legendre, 1991) and H. fossilis (Sundaranaj et al., 1982)
when fed at different periods of the day. In the present study, FCR and PER improved by
some 60% when fish were under a self-feeding regimen, this increase becoming significant
when FCR is considered. In rainbow trout, a higher feed conversion efficiency for growth
has been found when the fish are fed in phase with their natural feeding activity, a result
apparently related to increased protein synthesis and retention (Bolliet et al., 2001).
Better diet utilisation when fed at certain times of the day could be explained by
coincidence with natural rhythms of secretion, activation or synthesis of digestive and/or
metabolic enzymes. The intestinal protease activity in European sea bass has been
described as having a nychthemeral rhythm with increased activity at night, regardless
of the frequency of feeding or deprivation (Martı́nez-Bebiá et al., 1995). Boujard and
Leatherland (1992b) have detected changes in the concentration of several hormonal
metabolites in trout during the day, this being associated with feeding and/or photoperiod.
The adaptation of the feeding schedule to these metabolic, digestive and other rhythmic
processes related to the utilisation of nutrients (i.e. hormone release) appears to improve
FCR and PER indices when the fish are fed according to their natural rhythm of ingestion.
Diet chemical properties, including nutrient availability, are factors expected to affect
voluntary feed intake. It has been demonstrated that fish can discriminate between diets
differing in their macro- and micronutrient composition (Aranda et al., 2001). Sea bass can
also discriminate between diets with different methionine concentrations (Hidalgo et al.,
1988). Therefore, it is possible for fish to discriminate and adjust feed intake when facing
unbalanced amino acid diets.
According to the results, both soy-protein experimental diets (either methioninedeficient or free-methionine-supplemented) resulted in a similar behaviour to that for
controls, concentrating feeding demands during the evening (although feeding frequency
changed with respect to controls). In the case of the diet supplemented with free
methionine, the feeding period was longer, though the total daily number of requests
resembled that of the control diet and lower than the demand recorded for the methioninedeficient diet; however, in no case were the differences significant. The lower feed intake
of diets containing protein sources other than fishmeal might be attributed to essential
amino acid deficiencies, as shown for arginine (Kim et al., 1992a; Walton et al., 1986),
leucine (Choo et al., 1991) and methionine (Kim et al., 1992b). Essential amino acid
deficiency has been widely demonstrated to reduce feed intake, where normal intake
values are reached only when the amino acid concentration in the diet meets the
requirements of the fish, as reviewed by De la Higuera (2001). On the contrary, when
the European eel, Anguilla anguilla, was fed on diets containing sunflower protein
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supplemented with needed amino acids, feed intake and utilisation reached values similar
to those for fish fed on a fishmeal diet (Garcı́a-Gallego et al., 1998; De la Higuera et al.,
1999).
Soy meal is the vegetable-protein source most frequently used in practical diets to
replace fishmeal. Nevertheless, its methionine deficiency for most fish species limits
fishmeal-protein substitution. Attempts to correct this deficiency by adding free methionine to the diet have improved growth rates when compared to non-supplemented diets,
though growth rates remained lower than those for a control diet (Drabrowski and Kozak,
1979; Ketola, 1982; Murai et al., 1987). Gilthead sea bream responds to free methionine
supplements in the same way as other fish species: improving the nutritional parameters
with respect to unsupplemented soy meal but failing to reach fishmeal values (Sierra et al.,
1993).
Inefficiency of essential amino acid supplementation might be attributed to an
imbalance of postprandial amino acid availability due to the early absorption of dietary
free-supplemented amino acids. In this sense, in the European sea bass (Thebault, 1985),
trout (Cowey and Walton, 1988; Garzón, 1995), carp (Garzón, 1995) and gilthead sea
bream (Sierra, 1995) a postprandial imbalance was observed between protein and freesupplemented amino acids. In the case of gilthead sea bream fed on a soy-protein-based
diet supplemented with free methionine, the maximum concentration of plasma methionine was detected 2 h after ingestion, while protein amino acids reached maximum
concentrations 8 h after feed intake (Sierra, 1995). Nevertheless, when methionine
assimilation is delayed by microencapsulation with a protein coat, protein synthesis and
growth improve significantly, reaching values close to those for a fishmeal-control diet
(Sierra, 1995).
Results of the abovementioned works could be related to the lengthening described
above for the intake phase, to be interpreted as an adjustment of feeding behaviour to
correct postprandial imbalances after dietary amino acid uptake. The fish respond to an
earlier uptake of methionine by increasing their feeding-demand period, starting to feed 2
h earlier than the control. This change of feeding pattern would allow methionine from
most recent ingestion to coincide with protein amino acids from previous ingestion, so that
a sufficiently homogeneous amino acid pool is obtained at the protein-synthesis sites. Due
to the short duration of the second experiment (Table 1), we were unable to gather basic
data for growth rate and feed utilisation. Even though further long-lasting experiments are
needed, our preliminary results show that an appropriate feeding strategy might improve
dietary nutrient utilisation, self-feeding constituting an option for improving the utilisation
of alternative protein sources, especially when supplemented with the essential amino
acid(s) in which these sources are deficient.
Acknowledgements
The Dirección General de Investigación, Ministerio de Ciencia y Tecnologı́a, Spain,
Projects NMAR96-1881 and REN2000-1740-C05-03 RIES, supported this research. The
authors are grateful to Professor M. Jobling for his kindly help on the reading of this paper
and discussion of the results.
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101
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