January | February 2010

Feature title: Evaluation of Fishmeal Substitution

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The International magazine for the aquaculture feed industry

F: Process F: Process

the present study was to evaluate the

Table 2: Proximate analysis of the experimental diets (% as fed)
effect of partial or complete replace-
ment of fishmeal with increasing Diet
levels of plant protein origin like corn Chemical analysis
FM PPs/25 PPs/50 PPs/75 PPs/100

Evaluation of
gluten and soybean meal on growth
performance, feed utilization, body
composition and cost production of Moisture 8.50 8.00 8.30 8.70 8.80
sea bream fingerlings’ diets. Crude protein* 44.71 45.00 45.1 45.1 45.60

Fishmeal Substitution
Crude fat 17.56 16.42 15.32 14.48 13.32
Experimental protocol Crude fiber 1.14 1.95 2.57 2.00 2.18
Diet preparation - Five isocaloric Crude ash 9.30 8.00 6.55 4.14 2.33
and isonitrogenous diets were formu-
Nitrogen free extract 18.79 20.63 22.16 25.58 27.77
with selected Plant Protein Sources on Growth lated based on Fishmeal as the only
animal protein source or a mixture
Gross Energy (kcal/100gm) 1 495.15 493.59 490.06 496.18 497.05

Performance and Body Composition of gilthead of PPs (Corn gluten and Soybean P/E Ratio (mg protein/Kcal) 2 90.30 91.20 92.00 90.90 91.70

sea bream* Fingerlings meal) as plant protein sources

(Table 1). The diets formulated to be
1. Based on 5.64 Kcal/g protein, 9.44 Kcal/g fat and 4.11 Kcal/g carbohydrate (NRC, 1993).
2. Protein/Energy Ratio (mg Protein/Kcal).
almost containing 45% crude protein
by Abd Elhamid Eid* ,Badiaa Abd Elfattah*, Khaled Mohamed*
by replacing 25, 50, 75 and 100%
*Department of Animal & Fish Production, Fac. of Agric. Experimental methodology - and plasma albumin (Doumas, et al., 1977).
of the FM (fishmeal protein) in control
Suez Canal Univ. Ismailia 41522, Egypt The tested diets and faeces were analyzed Apparent protein digestibility was deter-
diet. Crystalline amino acids (L-lysine and
DL-methionine) were added to diets PPs for crude protein (CP %), ether extract mined using the method of Furukawa and

G
25, 50, 75 and PPs100% to become similar (EE %), crude fiber (CF %), ash (%) and Tasukahara (1966). For determination of
ilthead sea bream pro- essary, mainly through feeding, which aquaculture, as it would reduce depend-
to control diets. Fish oil and soybean oil moisture while whole body composition of protein digestibility the diets and faeces
duction in Mediterranean represents between 38 and 45% of ence on fish sources (Martinez-Llorens et
were added as dietary lipid sources (Table sea bream fish samples was also analyzed were collected during the last 15 days of
countries increased from operational costs (Lisac & Muir, 2000 al., 2009).
1). The diets were pelleted using a small except for crude fiber (CF %) according to the experimental period. Any uneaten feed
30,000 tons in 1996 to and Merinero et al., 2005). In the last decade, the increasing demand,
catering grinder with a 1.5 mm diameter the procedures
90,000 tons in 2005,which mean that price and world supply fluctuations of
and kept frozen until the experiment was described
sale prices dropped considerably, from Reductions in feeding costs can be fishmeal (FM) has emphasized the need
started. During the growth period (120 by A.O.A.C.
6.6€/kg in 1996 to 5€/kg in 2005, with obtained by optimizing feeding strate- to look for alternative protein sources in
days), each diet was randomly allocated to (1995) as
an historic minimum of 4€/kg in 2002 gies, nutrient levels in diets, and by using aquafeeds. Some plant ingredients have

Marine phospholipids
triplicate tanks of fish. Feed was offered by shown in Table
(APROMAR, 2006). To maintain the vegetable sources as substitutes for fish been studied in gilthead sea bream (lupin
hand at two meals / day (8:00h and 15:00) at 2 and Table 5.
profitability of gilthead sea bream oil and fishmeal. This aspect is also very seed meal, extruded peas and rapeseed
3% of body weight daily and the amount of The nitrogen
farms, cutting production costs is nec- important to improve the sustainability of meal) but Poaceae and Fabaceae seeds
diets were readjusted after each weighing. free-extract
and their by- products, among which
(NFE %) was A new generation of omega-3 lipids
corn gluten and soybean meal, in par- Experimental design - Sea bream
Table 1: Composition of the experimental diets
ticular, are widely used in fish nutrition
calculated by with a broader spectrum of health
fingerlings were obtained from a private fish
Diet because of their high protein content farm in Damietta governorate. Fish were
difference. benefits.
Ingredients (g/100g)* Blood samples
FM PPs/25 PPs/50 PPs/75 PPs/100 (40-60%), low cost and relative wide- acclimated to laboratory conditions for 2 were col- - High DHA contents, preferably in
spread availability. Therefore, soybean weeks before being randomly distributed into lected using
Fish meal (CP 68%) 63 47.24 31.52 15.78 - meal being the most nutritive and it fiberglass tank of 300-L water capacity each, heparinized
easily digestible and highly bio
Corn gluten meal (62%) - 9 20 45 62 is used as the major protein source in in Ashtom Elgamel, Port-Said governorate.The syringes from available form for aquaculture use.
Soybean meal (44%) - 13.7 24 13 15 many fish diets. Partial or even total water was obtained from channel comes from caudal vein of
Yellow corn 21.5 14.30 8.20 8.90 5.05 replacement of dietary fishmeal by Mediterranean sea. Fish of 10±0.2 g initial - Numerous benefits on improving
the experimen-
Fish oil+ Soya oil (1:1) 1** 12 12 12 12 12 soybean meal protein sources had suc- body weight were distributed into 15 experi- tal fish at the
the immune response, better
L-Lysine -- 0.26 0.62 1.69 2.32 cessfully accomplished with tilapia diets mental tanks in triplicate groups of 50 fish weight gain and physical
termination
DL- Methionine -- -- 0.16 0.13 0.13 (Fagbenro and Davies, 2002). Some each. The photoperiod was regulated to be
Vit & Min mix2 3 3 3 3 3 studies with gilthead sea bream have
of the experi- conditions of land animals.
12h light: 12h dark. Water temperature was ment. Blood
Cr2O33 0.5 0.5 0.5 0.5 0.5 shown that partial replacement of FM maintained at 25ºC by a 250- watt immersion was centrifuged
Total 100 100 100 100 100 by PPs is possible (Robaina, et al., 1995; heater with thermostat. Water temperature at 3000rpm for
Hassanen, 1997a, b; 1998; Kissil, et al., and dissolved oxygen were recorded daily (by
1- Mixture of fish oil and soybean oil (1:1 w/w). 5 minutes to
2000; Sitja-Bobadilla et al., 2005 and Metteler Toledo, model 128.s/No1242), other allow separa-
2- Each Kg vitamin & mineral mixture premix contained Vitamin A, 4.8 million IU, D3, 0.8 million Martinez-Llorens et al., 2009). Studies water quality parameters including pH and
IU; E, 4 g; K, 0.8 g; B1, 0.4 g; Riboflavin, 1.6 g; B6, 0.6 g, B12, 4 mg; Pantothenic acid, 4 g; tion of plasma
with sea bass have also reported some ammonia were measured every two days by
Nicotinic acid, 8 g; Folic acid, 0.4 g Biotin,20 mg, Mn, 22 g; Zn, 22 g; Fe, 12 g; Cu, 4 g; I, 0.4 which was
success to partial replacing of FM by pH meter (Orion model 720A, s/No 13062)
g, Selenium, 0.4 g and Co, 4.8 mg. subjected to
PPs (Lanari, 2005 and Tibaldi, et al., and ammonia meter (Hanna ammonia meter).
3- Cr2O3: Chromic Oxide determina- Fiskerihavnsgade 35 Phone +45 79120999
2006). Studies of using corn gluten to Water salinity was 34ppt. The average water tion of plasma P.O. Box 359 Fax +45 79120888
* obtained from the local market. feed carnivorous fish (sea bream) are quality criteria of all tanks are presented in total protein 6701 Esbjerg E-mail 999@999.dk
very limited; therefore, the scope of Table 3. All fish in each tank were weighed (Armstrong Denmark Web www.999.dk
every 10 days. and Carr, 1964)
*Sparus aurata

12 | InternatIonal AquAFeed | January-February 2010 January-February 2010 | InternatIonal AquAFeed | 13

999_AD_IAF0904V3.indd 1 22/06/2009 14:01
 F: Process F: Process

Table 3: Average water quality in an oven at 60oC and kept in airtight bream diets with no significant differences fish. In fish, protein digestibility
Table 5: Whole body composition (% fresh weight ) of see bream (S. aurata) fingerlings fed the
parameters in the experimental tanks containers for subsequent chemical analysis. (P≥0.05) in growth performance compared is generally high ranging from
experimental diets
used in the study. to the control (Table 4). This conclusion 75% to 95% and the apparent
Parameter Means ± SD Final digestible coefficient of proteins
Statistical analysis - All data of is in agreement with Gomes et al (1995a Chemical analysis Initial
growth performance, body composition & b) for rainbow trout. These workers FM PPs/25 PPs/50 PPs/75 PPs/100 from fishmeal is often higher
and blood parameters were analyzed by reported that replacement of fishmeal than 90% in salmonids (NRC,
Temperature (ºC) 25 ±1
one-way analysis of variance (ANOVA) by plant protein sources had no adverse 1993). Soybean meal contains
Oxygen (mg/L) 5.4 ±1 Moisture 70.50 59.91 a 60.44 a 60.75 a 63.50 b 64.33 b
using the general linear models procedure effects on growth. The optimal rate of sub- various anti-nutritional factors
Ammonia (NH3, mg/L) 0.011± 0.0001 Crude protein 14.25 17.56 a 17.50 a 17.56 a 17.34 b 16.90 b such as the anti-trypsin and an
of statistical analysis system (SAS) version stitution found in the present research was
pH 7.1 ± 0.10 8.02, (1998). Duncan's multiple range test closed with Lanari (2005), he reported Crude fat 10.5 15.62 a 15.70 a 15.77 a 14.00 b 13.90 b anti chimotrypsin factors, lectins,
(Duncan, 1955) was used to resolve dif- that soybean meal can substitute up to Crude ash 4.75 6.91a 6.36 a 5.92 b 5.16 c 4.87 d
oligosaccharides and a low level
Salinity (ppt) 34.0 ± 0.4
ferences among treatment means at 5% 25% of total protein of the sea bass diets of methionine. Corn gluten has
Values in the same row with a common superscript letter are not significantly different (P≥0.05) also a low level of amino acid
or faeces from each tank was carefully significant level using the following model. without any negative effect on growth
removed by siphoning about 30 min after performance. Higher value than reported lysine reduces the protein digest-
the last feeding. Faeces were collected Results & discussion in the present study was reported by Oliva-Teles, 2003 and Gómez-Requeni et These results of feed utilization related to ibility and amino acid availability of these
by siphoning separately from each repli- The present study indicates that PPs Gallagher (1994) in diets for hybrid striped al., 2004); short-term studies have shown apparent protein digestibility of diets used plant protein ingredients.
cate tank before feeding in the morning. (corn gluten and soybean meal) can replace bass, where soybean meal substituted 44% that at least 60-75% of FM can be replaced in the experiment which showed worst Corn gluten meal (CGM) is considered
Collected faeces were then filtered, dried 25% to 50% of fishmeal protein in sea of fishmeal without evidencing a negative by mixture of PPs without compromising feed utilization of sea bream fed on diets to have a good digestibility (NRC, 1993).
effect on the growth performance for these species. In containing high mixture of PPs (corn gluten Diets containing 20% of CGM meal had
Table 4: Growth performance and feed utilization of sea bream (S. aurata) fed the experimental diets feed intake and the present study, the effects of FM replace- meal and soybean meal) was possibly due to a very good digestibility, in accordance
he also report- ment were studied on growth performance the low biological value of such based diets, with the results of Morales et al.(1994)
Diet and feed utilization. This scenario, a high which are in agreement with Robaina, et al.,( and Gomes et al. (1995 a) in rainbow
Parameters ed that up to
FM PPs 25 PPs50 PPs75 PPs100 75% of fishmeal level of FM replacement by (50-75PPs %) 1995), Boonyaratpalin et al.,(1998), Regost trout fed diets containing about 20% corn
protein can be produced a slight reduction in growth et al.,(1999), Lanari (2005), Sitja-Bobadilla et gluten meal. In contrast, apparent digest-
replaced with performance. Concerning the results of al. (2005), and Tibaldi, et al.,(2006). ible coefficient of diets with high levels of
Average Initial body weight (g) 10.1±0.05 10.2±0.25 10.3±0.10 10.1±0.10 10.0±0.23 soybean meal. feed utilization in terms of FCR, PER and Regarding to feed digestibility (Table plant proteins was very low. In common
M o r e o v e r, FE in the present study, the same trend 4), several investigations were conducted carp, Pongmaneerat et al. (1993) observed
Average Final body weight (g) 102.6 a ±2.2 101.3 a ±0.3 97.7 b ±0.20 85.2 c ±0.2 78.9 d ±0.20
Sitja-Bobadilla was showed with growth performance. to evaluate PPs and their digestibility by that the apparent protein digestibility
91.1 a
Average Weight gain (g) 92.5 a ±1.1 87.4 b ±0.9 75.1c±0.10 68.9 d±1.10 et al., (2005)
±1.2
1.78 b reported that
SGR (% / d)1 1.93 a ±0.02 1.91 a ±0.01 1.87 a ±0.02 1.72 b ±0.01 up to 75% of
±0.09
151.93 a 152.51 a 153.15 a 159.62 b 159.30 b fishmeal pro-
Feed intake (g)
±0.4 ±0.2a ±0.10 ±0.2 ±0.10 tein can be The natural way.
Feed conversion ratio (FCR2 1.64 d ±0.10 1.67 d±0.1 1.75 c ±0.10 2.13 b ±0.1 2.31 a ±0.20 replaced by
plant protein
1.04 b
Protein efficiency ratio3 1.35 a ±0.01 1.32 a ±0.02 1.27 a ±0.01 0.95 b ±0.20 sources for
±0.10
juvenile sea

Biomin P.E.P.
Feed efficiency4 0.61 a ±0.1 0.60 a ±0.10 0.57 a ±0.10 0.47 b±0.10 0.43b ±0.12
Sponsor of the
®
bream, which
2010
HSI (%)5 3.2 a ±0.1 2.97 a ±0.1 2.93 a ±0.12 2.71b ±0.01 2.56 c ±0.12 also is in agree- World Aquaculture
st an d
Apparent Protein Digestibility (APD)6 88.25 a ±0.3 87.39 a ±0.2 86.09 a ±0.1 73.16 b±0.2 65.32 c ±0.1
ment with the Visit us at our
present study
Powerful phytogenics at work 129-131
PTP (g/dl)7 5.21±0.10 5.20±0.12 5.15±0.10 5.03±0.12 5.01±0.10 for sea bream

PA (g/dl)8 2.15±0.11 2.17±0.11 2.17± 0.12 2.07±0.02 2.08±0.08

fingerlings. In
the recent
improve feed efficiency.
PTG (g/dl)9 3.06±.0.12 3.03±0.10 2.98±0.11 2.96±0.09 2.93±0.01 years, signifi-
cant amount of
Survival rate (%)10 100 100 98 96 94
research has Biomin® P.E.P. is made with a unique
Values in the same row with a common superscript letter are not significantly different (P≥0.05). been con- blend of essential oils and prebiotics to provide
ducted on the
Specific growth rate = (100 x [(Ln final wt (g) – (Ln initial wt (g) / days.] a synergistic formula.
replacement of
Feed conversion ratio (FCR) = feed intake (g) / body weight gain (g).
FM by different It is designed specifically to support digestion
Protein efficiency ratio (PER) = gain in weight (g) / protein intake (g). PP sources. and improve feed conversion.
Feed efficiency = body weight gain (g) / feed intake (g). In European
sea bass
Hepato-somatic index = 100 x liver wt / fish wt.
( D. l a b r a x )
6 - Apparent protein digestibility, APD (%) (Kaushik et
7 - Plasma Total Protein, PTP (g/dl) al., 2004)
and Gilthead For enquiry, please e-mail
8 - Plasma albumin, PA (g/dl)

www.aqua.biomin.net
sea bream
9 - Plasma total globulins= plasma total protein- plasma albumin, PTG (g/dl) (S.aurata) aqua@biomin.net
10 - Survival rate =No of survive fish/total No. of fish at the beginning X100 (Pereira and
Ad_PEP_IntAquafeed_FZE_12_09.indd 1 16.12.2009 16:30:59 Uhr
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 F: Process
had to be near 94% in a diet without
fishmeal (corn gluten meal, soybean and
meat meal).
Results of apparent protein digest-
ibility in the present study recorded that
the dietary inclusion of high levels of
corn gluten and soybean meal in replace-
ment of fishmeal led to a significant
decrease in protein digestibility which
are in agreement with Lanari (2005),
Tibaldi et al. (2006) and Sampaio-Oliveira
and Cyrino (2008). The value of hepato-
somatic index was found to be similar to
that reported for sea bass by Ballestrazi
et al., (1994) and Dias et a. , (1998),
Table 6: Feed cost (L.E) for producing one Kg weight gain by sea bream (S. aurata) fingerlings fed on the
experimental diets
Experimental Cost Relative
diets (L.E)/kg fishmeal diets
FM 6.56 100
PPs25 5.66 86.29
PPs50 4.77 72.71
PPs75 3.96 60.36
PPs100 3.12 47.56
The local market price were 8LE for fish meal, 2.50LE for gluten, 1.70LE for soybean meal, 1.00 LE for
yellow corn, 9 LE for oil, 5 LE for Vit. & Min.
they reported that the values of HSI
were 2–3% or above. Effect of the
experimental diets on hepato–somatic
index confirmed that the fish fed on diets
containing high levels of corn gluten
meal and soybean meal evidenced a
significant (P≤0.05) decrement of the HSI
in relation to the utilization of glycogen,
stored as an energy source.The results
are in agreement with Lanari (2005) and
Sampaio-Oliveira and Cyrino(2008).
Effects of the experimental diets on
whole body protein concentration (Table
5) were very small with exception of fish
diet containing FM, 25 and 50%PP which
showed a significant difference (P≤0.05)
compared to the other experimental
diets (75 and 100%PP). Fish body fat
content decreased with increasing level
of PPs substitution. The low percentage
of fat stored with diets containing high
level of PPs is due to the limited ingestion
of the feed or to probable use of the
body fat as energy source and may be
also related to the carbohydrate levels
and type of the diets. These results are
in agreement with Lanari (2005), Tibaldi
16 | InternatIonal AquAFeed | January-February 2010
et al.(2006) and Sampaio-Oliveira and
Cyrino (2008) for sea bass D. labrax and
Peres and Oliva-Teles (2009) for sea
bream S. aurata.
Calculation of the economical effi-
ciency of the tested diets was based
on the costs of feed because the
other costs were equal for all studied
treatments.
As described in Table 6 feed costs (L.E)
were the highest for the fishmeal diet and
gradually decreased with increasing the
replacing levels of plant protein sources.These
results indicate that incorporation of PPs in
sea bream diets reduced the total feed costs.
Feed cost
Decrease in
FCR (L.E/Kg)
feed cost (%)
weight gain
0.00 1.64 10.76
13.71 1.68 9.51
27.29 1.89 9.02
39.64 2.13 8.43
52.44 2.31 7.21
However, high replacing levels of fishmeal by
PP (75 and 100%PP) adversely affected all
the growth and feed utilization parameters
(Table 4), but the incorporation of PPs in
sea bream diets seemed to be economic
as incorporation of PPs in the diets sharply
reduced feed costs by 13.71, 27.29, 39.64 and
52.44% for 25PPs, 50PPs, 75PPs and 100%,
respectively. The reduction of feed costs was
easily observed for the feed costs per Kg
weight gain which decreased with increasing
incorporation levels of PPs in agreement with
Soltan (2005) for Nile tilapia and Eid and
Mohamed (2007) for sea bass fingerlings.
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Egyptian J. Nutrition and Feeds,1(special Issue
):335-348.
Hassanen, G.D.I. (1997b). Effect of diet composition
and protein level on growth, body composition and
cost of production of gilthead sea bream (Sparus
aurata). Egyptian .J. Aquat. Biol & Fish., 2: 1-18.
January-February 2010 | InternatIonal AquAFeed | 17
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