Aquaculture Nutrition

doi: 10.1111/j.1365-2095.2007.00523.x 2008 14; 242248

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1
1

Departamento de Zootecnia/Nao Ruminantes, Setor de Piscicultura, Universidade de Sao Paulo, Sao Paulo; 2 Centro de Ciencias Agrarias, Biologicas e Ambientais, Universidade Federal do Reconcavo da Bahia, Bahia; 3 Departamento de Agroindustria, Alimentos e Nutricao, Universidade de Sao Paulo, Sao Paulo, Brazil

Using the sh silage to partially replace proteic feedstu in aquafeeds is an alternative to mitigate sanitary and environmental problems caused by the lack of adequate destination for sheries residues. It would also lower feed costs, consequently improving sh culture protability. However, using sh silages in aquafeeds depends on determination of its apparent digestibility coecients (ADC). This work aimed to determining the ADC of crude protein and amino acids of acid silage (AS), biological silage (BS) and enzymatic silage (ES) for juvenile Nile tilapia (94.5 12.7 g). The ADCCP was: 92.0%, 89.1% and 93.7% for AS, BS and SE respectively. The average ADC of amino acids was: 91.8%, 90.8% and 94.6% for AS, BS and ES respectively. Results encourage the use of AS, BS and ES to partially replace protein sources in balanced diets for neotropical sh.
KEY WORDS:

alternative ingredient, amino acids, by-products, digestibility, sh silage, Oreochromis niloticus

Received 30 January 2007, accepted 7 August 2007 Correspondence: Ricardo Borghesi, Departamento de Zootecnia/Nao ruminantes, Setor de Piscicultura, Escola Superior de Agricultura Luiz de Queiroz, Universidade de Sao Paulo, PO Box no. 9, 13418-900 Piracicaba, Sao Paulo, Brazil. E-mail: ricardo_borghesi@yahoo.com.br

The growth of aquaculture, associated to the intensication of production systems, has increased the demand for alternative, high-quality feedstu (Espe et al. 1999), which would allow the formulation and use of nutritionally adequate, economically viable, and environmentally-friend diets. Because of its biological value, appropriate amino

acids prole and adequate fatty acids contents, not to mention its attractiveness to sh, sh meal (FM) is considered the main protein source for formulation and manufacturing of sh diets (Sales & Britz 2003). The increasing demand and progressive scarcity of FM in the international market boosted its price and launched the quest for reduction of FM in sh diets and the consequent search for alternative, acceptable and digestible protein sources (Pereira & Oliva-Teles 2003; FAO 2004; Cyrino et al. 2006). Knowing nutrient digestibility of feed ingredients elicits interchangeability of ingredients without reducing animal performance. In combination, chemical analysis and apparent digestibility coecient (ADC) results allow to precisely estimate not only the contribution of a particular protein source to a complete sh feed, but also how much feed wastes and undigested nutrient (faeces) will potentially accumulate in sh ponds (Furuya 2001; Koprucu & Ozdemir 2005). Fisheries by-products and rejects may accumulated and cause environmental, health and economical problems; but when processed and turned in by-products, they can become alternative, potentially good feedstu for aquafeeds which has been considered to be the case of sh silage, according to Vidotti et al. (2002, 2003) and Carvalho et al. (2006) is a product of high biological value. Fish silage is a liquid or semi-liquid product resulting from the preservation (ensilage) of whole sh or parts by the addition of acids (acid silage, AS) or carbohydrate-induced microbial fermentation (biological silage, BS). In the ensilage process, proteins are hydrolysed by naturally present and (or) added enzymes (enzymatic silage, ES); the process is favoured by the adjustment of pH. The nal result is a paste-like product, rich in protein, short-chain peptides and free amino acids (Morales-Ulloa & Oetterer 1997).

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Several authors have demonstrated the nutritional and economical feasibility of using sh silage in diets of several sh species, not to mention environmental benets of this practice (Fagbenro & Jauncey 1995a,b; Goddard & Al-Yahyai 2001; Vidotti et al. 2002; Goddard & Perret 2005). But is important to know that in the Europe, the international feed legislation does not allow to include products/ingredients of sh in feed to same species. The objective of this work was thus determining the ADC of protein and amino acids present in AS, BS and ES obtained from Nile tilapia (Oreochromis niloticus) processing by-products and trash sh, to Nile tilapia itself.

Crude protein of silages and diets were determined by microKjedahl method (AOAC 1984). The chromic oxide contents were quantied by atomic absorption spectrophotometry, after acid digestion. Amino acid contents of silages and diets were determined by liquid chromatography in an autoanalyser; samples were hydrolysed with HCl 6 N for 22 h at 110 C (Moore & Stein 1963). Tryptophan was determined after enzymatic hydrolysis with pronase at 40 C for 24 h, followed by colorimetric reaction with 4-dimethyl-aminobenzaldehyde in 21.2 N sulphuric acid; reading was performed at 590 nm and tryptophan contents calculated from a standard curve (Spies 1967).

The raw material was 20% of Nile tilapia processing (lleting) by-products heads, viscera, scales, ns, vertebral column, skin, adhered tissues plus 80% whole Nile tilapias (trash sh), grinded in an electrical mincer (ML-4.0 WEGlline, Acao Cientica, Piracicaba, Brazil) to yield 60 kg of a homogenous mass.

The homogenous mass obtained was split into six plastic containers (10 kg/container) and added of 200 ppm of butylhydroxy-toluene dissolved in ethylic alcohol plus 3% of a 1 : 1 mixture (volume/weight) of 88% formic acid and 100% propionic acid. After homogenous acidication (hand mixing) of the mass, the containers were covered with plastic lids and set to rest at room temperature. AS became a pretreatment for other silages. Two containers were reserved as AS, and incubated for 4 weeks; the others were used to produce the BS and ES. Biological silage was prepared from the AS 3 days after incubation, when the supernatant lipid layer was removed. To obtain the BS, 14 000 mg kg)1 of inoculum solution of Lactobacillus plantarum plus 18% of sugarcane molasses (volume/weight) were added and homogenized to AS containers, which were covered with plastic lids and kept at room temperature for 3 weeks. Enzymatic silage was obtained by the addition of 10 g of protease type II from Aspergillus oryzae (0.13 unit mg)1 solids; Sigma Chemical Corporation, Saint Louis, Missouri, USA) diluted in 100 mL of distilled water to AS containers, also 3 days after incubation. After hand homogenization, the containers were covered with plastic lids and kept at room temperature for 1 week.

The determination of ADC of silages was carried out by the indirect method, using 1.0 g kg)1 chromic oxide III (Cr2O3) as dietary marker. The semi-puried reference diet (RD) was formulated based on albumin and gelatin protein (Table 1). The test diets were obtained by replacing 30% of RD with the tested silages respectively. To be used in diets formulation and processing, studied silages were oven-dried (50 C; 24 h), grinded in hammer mill, and required amounts were weighed and homogenized to the feedstus mix which was added of water (45 C) at the

Table 1 Composition of reference diet (as fed basis)

Ingredient Albumin Gelatin Corn starch Soybean oil a-Cellulose Di-calcium phosphate Mineral and vitamin mix1 Vitamin C2 Sodium chloride BHT Chromic oxide
1

g kg)1 320.00 77.00 44.13 60.00 60.00 30.00 5.00 0.50 5.00 0.20 1.00

BHT, butyl-hydroxy-toluene. Mineral and vitamin mix (SupreMais): vitamin A 1 200 000 IU; vitamin D3 200 000 IU; vitamin E 12 000 mg; vitamin K3 2400 mg; vitamin B1 4800 mg; vitamin B2 4800 mg; vitamin B6 48 000 mg; vitamin B12 4800 mg; folic acid 1200 mg; calcium pantothenate 12 000 mg; vitamin C 48 mg; biotin 48 mg; choline 65 mg; nicotinic acid 24 000 mg; Fe 10 000 mg; Cu 600 mg; Mn 4000 mg; Zn 6 000 mg; I 20 mg; Co 2 mg; Se 20 mg. 2 Vitamin C: calcium salt, 2-monophosphate ascorbic acid, 42% of active principle.

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rate of 20% of the total diet weight. The mixture was extruded through a mincer (ML-4.0 WEG-lline); pellets were collected, dried overnight in a forced air oven (55 C), grinded to 25 mm pellets, sized, hermetically packed and stored under refrigeration until use. The temperature and the oxygen of the feeding and digestibility tanks were maintained by heaters (26 C) and the porous rock was coupled to a central air pump. Juvenile Nile tilapias (94.54 12.66 g) were stored in eight cylindrical, plastic cages (80-L; mesh size 1.5 cm; 16 sh/cage) installed in eight 250-L circular feeding tanks. Fish were hand-fed ad libitum four daily meals (08:0017:30) and cages were then transferred and kept overnight into four, 300-L cylindrical-conical faeces collection tanks with a recirculation system (Pezzato et al. 2002). Feeding tanks were supplied by continuous ow, water recycling system; faecescollecting tanks were equipped with individual water recycling systems. Water temperature was maintained at 26 1 C in both systems by the use of electrical heaters coupled with thermostats. Feces were collected by sedimentation to collecting bottles connected to the bottom of the tanks, oven-dried (55 C; 24 h), grinded and stored under refrigeration ()15 C) for posterior analysis. The ADC of nutrients of the tested ingredients and RD were calculated according to the following equation Pezzato et al. (2002):    % Cr2 O3 diet % nutrient faeces ADC100 100 % Cr2 O3 faeces % nutrient diet  where: ADC, apparent digestibility coecient (%); % Cr2O3 diet, chromic oxide percentage in diet; % Cr2O3 faeces, chromic oxide percentage in faeces. As the tested ingredients substituted 30% of the RD, the apparent digestibility of the ingredients was calculated separately from nutrients according to the following equation: ADCn ADCTD ADCRD y z 

No rejection of diets containing AS, BS and ES was registered. Fish silage contains considerable amounts of free amino acids released in the hydrolysis process which may have acted as attractants to sh (Berge & Storebakken 1996; Furuya 2001). Table 2 shows the amino acids composition and crude protein contents of the ingredients, while Table 3 shows the amino acids composition and crude protein contents of the RD and test diets. Values of ADC of amino acids (ADCAA) and protein of the silages are presented in Table 4. Values of ADC of crude protein (ADCCP) were lower than those registered by Goddard & Al-Yahyai (2001) for AS fed to Nile tilapia, and Stone et al. (1989) who also fed AS to rainbow trout (Oncorhynchus mykiss), and recorded ADCCP values of 95.1 and 94% respectively. On the other hand, values of ADCCP found in this research were superior to those registered by Vidotti et al. (2002) for pacu (Piaractus mesopotamicus) fed both AS and BS (74.7% and 83.4%, respectively), and by Hardy et al. (1984) who used AS as protein source in rainbow trout feeds (80.1%). Studying the digestibility of AS added as feed ingredient in rainbow trout diets, and using the manual extrusion method of faeces collection, Stone & Hardy (1986) found an ADCCP

Table 2 Amino acids and crude protein contents of the acid, biological and enzymatic silages (mg g)1 dry weight)
Silages Amino acids Essential Valine Methionine Isoleucine Leucine Threonine Phenylalanine Lysine Histidine Arginine Tryptophan Nonessential Cystine Tyrosine Aspartic acid Serine Glutamic acid Proline Glycine Alanine Crude protein2
1 2

Acid 27.01 21.7 32.3 35.0 20.4 20.5 33.3 14.0 29.0 3.6 7.7 13.0 54.6 18.3 60.1 28.0 44.1 37.6 542 5.0

Biological

Enzymatic

14.2 18.6 12.0 24.1 16.4 13.1 24.1 9.9 18.6 2.4 6.6 9.9 40.5 10.9 36.1 24.1 26.3 23.0 330 2.6

29.8 22.1 22.1 33.1 21.0 22.1 32.2 13.2 29.8 4.0 7.8 13.2 55.1 18.7 69.5 28.8 43.0 41.9 545 3.0

where: ADCn, apparent digestibility coecient of nutrients; ADCTD, apparent digestibility coecient of nutrients in test diet; ADCRD, apparent digestibility coecient of nutrients in reference diet; y, reference diet proportion; z, test diet proportion.

Recorded data were submitted to ANOVA and Turkeys test (P 0.05) by the statistical software SAS (SAS 2000).

Average of duplicates Average of triplicates SE.

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Table 3 Amino acids and crude protein contents of the RD and test diets (mg g)1 dry weight)
Diets Amino acids Essential Valine Methionine Isoleucine Leucine Threonine Phenylalanine Lysine Histidine Arginine Tryptophan Nonessential Cystine Tyrosine Aspartic acid Serine Glutamic acid Proline Glycine Alanine Crude protein2 RD 18.71 15.6 16.7 26.0 11.8 15.6 21.9 6.8 19.8 4.9 6.5 8.3 29.5 19.2 62.1 22.1 23.3 22.9 377 2.2 RD + AS RD + BS RD + ES

21.17 17.41 21.36 28.67 14.37 17.05 25.30 8.95 22.54 4.51 6.85 9.70 37.00 18.91 61.44 23.85 29.52 27.29 426 1.8

23.13 20.75 26.18 31.70 19.18 18.26 30.51 12.76 25.85 3.24 7.36 12.06 50.32 16.06 52.84 26.80 38.72 33.18 478 2.0

18.87 19.63 15.02 26.78 17.76 15.79 26.51 10.88 21.94 2.88 6.95 10.88 44.84 13.23 46.08 25.49 31.28 28.65 394 1.3

RD, reference diet; AS, acid silage; BS, biological silage; ES, enzymatic silage. 1 Average of duplicates. 2 Average of triplicates SE.

value of 88.7%, which is close those registered for ADCCP of BS in this research. Fagbenro & Bello-Olusoji (1997) found ADCCP values of 90.8%, similar to those obtained in this research for AS and BS, when feeding BS and soybean meal-based diet (1 : 1 ratio) to African catsh. The authors used the dissection method for faeces collection and chromic oxide as a marker. This research has, in particular, quality of the raw material, processing method, mineral (ash) and lipids contents of silages, and storage period as dierentials which may have yielded higher ADC values (Anderson et al. 1992; NRC 1993; Allan et al. 2000; Lee 2002). Actually, proteins originated from bones and connective tissues are less digested than those originated from muscles (NRC 1993). In addition, the high ADCCP values registered conrm Nile tilapias ability of digesting animal protein, corroborating results reported by Hanley (1987) and Masumoto et al. (1996). Values of ADCAA of the silages and the RD are presented in Table 4. Except for tryptophan, phenylalanine and aspartic acid, all other ADCAA of amino acids presented dierences (P = Fr < 0.05). Average values for ADCAA were 90.76%, 91.83% and 94.61%, for BS, AS and ES respectively. The tested ingredients presented higher AD CAA values, nearing those found by Koprucu & Ozdemir (2005) studying the digestibility of anchovy meal (91.2%),

Table 4 Apparent digestibility coefcients of essential amino acids, nonessential amino acids, and crude protein of the reference diet and the acid, biological, and enzymatic silages

Silage Reference diet (%) 97.2a 97.0ab 96.2ab 96.3a 95.1ab 96.7a 97.2a 97.6a 98.2a 97.9a 97.1a 95.1b 97.0a 96.5a 97.8a 97.6a 96.7a 97.0a 96.7a Acid (%) 92.8ab 94.8a 97.2a 91.2b 91.7b 93.0a 94.8ab 96.2ab 89.5b 97.9a 92.1b 95.8ab 94.4a 90.4ab 89.5ab 84.5b 81.0b 86.3b 92.0c Biological (%) 89.0b 96.0ab 89.3b 91.5b 94.3ab 91.0a 94.4b 94.9b 88.9b 97.0a 93.7ab 94.7b 93.8a 86.8b 88.6b 83.8b 80.9b 85.1b 89.1d Enzymatic (%) 97.3a 98.3b 96.3ab 95.6ab 96.9a 96.0a 97.1a 97.4a 93.3b 97.7a 97.2a 97.3a 95.7a 90.7ab 93.5ab 87.0b 82.8b 93.0ab 93.7b

Amino acids Essential Valine Methionine Isoleucine Leucine Threonine Phenylalanine Lysine Histidine Arginine Tryptophan Nonessential Cystine Tyrosine Aspartic acid Serine Glutamic acid Proline Glycine Alanine Crude Protein

SE

ANOVA

P > Fr

0.3 0.3 0.9 1.1 1.2 0.7 0.9 0.6 0.6 0.9 1.8 0.7 1.1 2.3 1.5 0.6 1.4 0.8 0.4

1.0 0.8 2.3 1.0 0.7 2.3 0.6 0.3 0.5 0.9 0.8 0.7 1.6 1.8 1.5 1.8 1.7 2.8 0.5

2.5 1.0 1.8 1.1 1.4 2.9 0.4 0.6 1.7 1.0 0.7 1.7 1.3 2.6 2.4 2.0 1.4 3.0 1.4

0.5 2.0 2.0 1.4 1.6 1.3 0.8 0.9 0.6 0.8 1.3 1.0 2.5 1.9 2.1 0.9 1.4 1.7 0.2

2.2272 0.9122 3.4484 1.2795 1.5076 3.6513 0.4019 0.2785 1.2801 0.3227 1.7344 0.0277 3.3335 5.2749 4.5161 2.2694 3.1116 5.2045 0.3965

0.0035 0.0305 0.0158 0.0057 0.0227 0.0667 0.0062 0.0046 0.0005 0.0939 0.0167 0.0036 0.3962 0.0258 0.0164 0.0003 0.0002 0.0055 0.0001

Average of triplicates SE; similar superscript letters in the column do not differ (P 0.05).

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soybean meal (87.1%) and corn gluten meal (89.6%) to Nile tilapia. Tryptophan presented the highest ADC among essential amino acids (EAA). These results oppose to those presented by Fagbenro & Bello-Olusoji (1997), working on digestibility of sh silage for the African catsh, and Portz & Cyrino (2004) regarding digestibility of soybean and FM for largemouth bass (Micropterus salmoides); in both cases, tryptophan presented the lowest ADC values. On the other hand, results agree with those reported by Anderson et al. (1992) regarding digestibility of soybean and FM, and Anderson et al. (1995), regarding digestibility of herring and anchovy meal, both for Atlantic salmon Salmo salar. Arginine had the lowest values of ADC among the EAA. These results concur with those of Hossain & Jauncey (1989) for FM in the feeding of common carp, but go up against those presented by Furuya et al. (2001), who studied digestibility of animal and vegetal ingredients for Nile tilapia, and Anderson et al. (1992) who studied the use of canola meal in diets for Atlantic salmon. Among the nonessential amino acids (NEAA), tyrosine presented the highest ADC, concurrent with the results reported by Portz & Cyrino (2004) regarding digestibility of sh and poultry viscera meals by largemouth bass. The lowest ADC among the NEAA was that of glycine. Hossain & Jauncey (1989) and Koprucu & Ozdemir (2005) working with FM, Anderson et al. (1992) working with FM and soybean meal, and Allan et al. (2000) working with meat and bones meal in diets for Nile tilapia, Atlantic salmon, and carp, respectively, also reported that among NEAA, arginine is the one which presents the lowest ADC. The lowest ADC of nutrients and amino acids of the BS may be connected to the amount of molasses added to produce this silage, for during the faeces collection period, shfed diets containing BS excreted larger amount of faeces which turned the water of the faeces collection tanks of a brownish colouration. Nile tilapia can use dietary carbohydrates rather eciently (Tengjaroenkul et al. 2000); however, excess dietary carbohydrates can negatively aect the digestibility of some nutrients by the species (Storebakken et al. 1998). Average ADCAA for AS, BS and ES reported in this study were higher than those reported by Furuya et al. (2001) for FM in the feeding of Nile tilapia, and by Portz & Cyrino (2004) for FM with largemouth bass, but lower than those reported by these last authors when working with soybean meal. Dierences in the use of amino acids from varying feedstus were also reported by Allan et al. (2000) for the silver perch (Bydianus bydianus).

Discrepancies in ADC in the protein and amino acids may be related to species-specic biological traits, i.e. its ability of using some nutrients, the faeces collection method, type of RD and marker, chemical composition of the ingredients, and feed processing technique (Anderson et al. 1992; NRC 1993; Allan et al. 2000; Lee 2002). The average values of ADCCP and ADCAA of the studied ingredients were rather similar; comparable results were reported by Portz & Cyrino (2004) for largemouth bass, Sales & Britz (2003) for South African abalone (Haliotis midae L.), Koprucu & Ozdemir (2005) for Nile tilapia and Hossain & Jauncey (1989) for carp. On the other hand, Anderson et al. (1992) did not nd proximate values of ADCCP and ADCAA for the same feedstu when working with salmon. At any rate, in spite average values of ADCCP and ADCAA being usually co-related, determining and knowing individual digestibility values of amino acids is very important to formulate complete diets, if maximum performance and avoidance of dietary deciency or excess of a certain amino acid is the ultimate goal (Anderson et al. 1992, 1995; Stone et al. 2000; Furuya et al. 2001; Lee 2002; Sales & Britz 2003). Table 5 shows values of protein and digestible essential and NEAA of RD, AS, BS and ES (mg g)1 dry weight). Table 6 elicits comparing values of EAA contents in the

Table 5 Values of protein and digestible essential and nonessential amino acids of the reference diet and of the acid, biological and enzymatic silages (mg g)1 dry weight)
Reference diet 18.21 15.1 16.1 25.0 11.2 15.1 21.3 6.6 19.4 4.8 6.3 7.9 28.6 18.5 60.8 21.6 22.5 22.2 364 Silage Acid Biological Enzymatic

Amino acids Essential Valine Methionine Isoleucine Leucine Threonine Phenylalanine Lysine Histidine Arginine Tryptophan Nonessential Cystine Tyrosine Aspartic acid Serine Glutamic acid Proline Glycine Alanine Digestible protein
1

25.1 20.6 31.4 31.9 18.7 19.1 31.6 13.5 25.9 3.5 7.1 12.5 51.5 16.5 53.8 23.7 35.7 32.5 499

12.6 17.9 10.7 22.1 15.5 11.9 22.7 9.4 16.5 2.3 6.2 9.4 38.0 9.5 32.0 20.2 21.3 19.6 294

29.0 21.7 21.3 31.6 20.4 21.2 31.3 12.9 27.8 03.9 07.6 12.8 52.7 17.0 64.9 25.1 35.6 39.0 510

Values calculated based on Table 2 and Table 3.

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Table 6 Values of essential, digestible amino acids of the silages and essential amino acids requirements for Nile tilapia, according to the NRC (1993)
Silages Acid Biological Enzymatic NRC (1993) (mg g)1 DP) (mg g)1 DP) (mg g)1 DP) (mg g)1 DP) 50.3 55.5 62.9 63.9 37.5 63.3 63.3 27.0 51.9 7.0 42.9 82.0 36.4 75.2 52.7 72.4 77.2 32.0 56.1 7.8 56.8 57.4 41.7 61.9 40.0 66.6 61.3 25.3 54.5 7.6 28.0 26.8 31.1 33.9 37.5 37.5 51.2 17.2 42.0 10.0

Amino acids Valine Methionine + cystine Isoleucine Leucine Threonine Phenylalanine + tyrosine Lysine Histidine Arginine Tryptophan

` Authors are indebted to Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (Sao Paulo State Research Foundation FAPESP) and to Coordenacao de Aperfeicoamento de Pessoal de N vel Superior (Coordination for Improvement of Superior Education Personnel CAPES) for research funds and scholarships granted.

DP, digestible protein; CP, crude protein.

silages with the EAA requirements of Nile tilapia as established by the NRC (1993). All three silage types have tryptophan as limiting amino acid. However, this result does not lessen the nutritional value of the silages, as any of them can or will ever be used as sole protein source in a balanced aqua feed. The partial dehydration of the silage (co-drying) through addition of other products or by-products is a tool that has been successfully used by many authors (Fagbenro 1994; Fagbenro & Jauncey 1995a; Fagbenro & Bello-Olusoji 1997). The mixture of sh silage and soy meal has been showing the best results in experiments of performance and digestibility with several species of sh. The co-drying of silages helps decreasing their moisture contents and spare potential limitations in their amino acids prole. For instance, Portz & Cyrino (2004) found that soybean meal contains 13 g of digestible tryptophan 100 g)1 of digestible protein for the carnivore largemouth bass. This value exceeds Nile tilapias nutritional requirement of tryptophan as xed by NRC (1993). Therefore, mixing soybean meal with sh silage would improve the amino acids prole and, consequently, the nutritional value of any aqua feed based on these feedstus. Acid silage, BS and ES were eciently used and presented high values of ADC of protein and amino acids for Nile tilapia. Therefore, they can be recommended as a partial substitute of FM in diets for this or any other sh species. ES and AS yielded generally better results than the BS. As the elaboration of AS production is simpler than the others, it would be recommendable obtaining and using this form silage in sh diets.

Allan, G.L., Parkinson, S., Booth, M.A., Stone, D.A.J., Rowland, S.J., Frances, J. & Warner-Smith, R. (2000) Replacement of sh meal in diets for Australian silver perch, Bidyanus bidyanus: I digestibility of alternative ingredients. Aquaculture, 186, 293310. Anderson, J.S., Lall, S.P., Anderson, D.M. & Chandrasoma, J. (1992) Apparent and true availability of amino acids from common feed ingredients for Atlantic salmon (Salmo salar) reared in sea water. Aquaculture, 108, 111114. Anderson, J.S., Lall, S.P., Anderson, D.M. & Mcniven, M.A. (1995) Availability of amino acids from various sh meals fed to Atlantic salmon (Salmo salar). Aquaculture, 138, 291301. Association of Ocial Analytical Chemists (AOAC) (1984) Ocial Methods of Analysis, 14th edn. AOAC, Washington, D.C., pp. 152160. Berge, G.M. & Storebakken, Y. (1996) Fish protein hydrolyzate in starters diets for Atlantic salmon (Salmo salar) fry. Aquaculture, 145, 205212. Carvalho, G.G.P., Pires, A.V.J., Veloso, C.M., Silva, F.F. & Carvalho, B.M.A. (2006) Silagem de res duo de peixe em dietas para alevinos de tilapia-do-nilo. Rev. Bras. Zootec., 35, 1. Cyrino, J.E.P., Bicudo, A.J.A., Sado, R.Y., Borghesi, R. & Dairiki, J.K. (2006) Producao de peixes e meio ambiente a busca pela denicao, formulacao e uso de alimentos ambientalmente corretos em piscicultura. In: 2nd Congresso Latino Americano de Nutrica o Animal (Colegio Brasileino de Nutricao Animal eds), p. 28. Anais. CLANA, Sao Paulo, Brazil, CD-ROM. Espe, M., Sveier, H., Hogoy, I. & Lied, E. (1999) Nutrient absorption and growth of Atlantic salmon (Salmo salar L.) fed sh protein concentrate. Aquaculture, 174, 119137. Fagbenro, O. (1994) Dried fermented sh silage in diets for Oreochromis niloticus. The Israeli J. Aquac., 46, 3, 140147. Fagbenro, O. & Bello-Olusoji, O.A. (1997) Preparation, nutrient composition and digestibility of fermented shrimp head silage. Food Chem., 60, 489493. Fagbenro, O. & Jauncey, K. (1995a) Water stability, nutrient leaching and nutritional properties of moist fermented sh silage diets. Aquac. Eng., 14, 143153. Fagbenro, O. & Jauncey, K. (1995b) Growth and protein utilization by juvenile catsh (Clarias gariepinus) feed dry diets containing co-dried lactic-acid-fermented sh-silage and protein feedstus. Bioresou. Technol., 51, 2935. FAO (2004) El Estado Mundial de la Pesca y la Acuicultura 2004. FAO, Roma, p. 168. Furuya, W.M. (2001) Alimentos ambientalmente corretos para piscicultura. In: 38th Reunia Anual da Sociedade Brasileira de Zoot o (Mattos, W.R.S., Cyrino, J.E.P. & Verdade, L.M. eds), pp. 515 527. Anais. FEALQ, Piracicaba, Brazil. Furuya, W.M., Pezzato, L.E., Pezzato, A.C., Barros, M.M. & Miranda, E.C. (2001) Coeciente de digestibilidade e valores de

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2007 Blackwell Publishing Ltd Aquaculture Nutrition 14; 242248