Distillers' dried grains with soluble in diets for Pacu, Piaractus mesopotamicus juveniles: Growth performance, feed utilization, economic viability, and phosphorus release

Journal Pre-proof Distillers’ dried grains with soluble in diets for Pacu, Piaractus mesopotamicus juveniles: growth performance, feed utilization, economic viability, and phosphorus release Katia R.B. Oliveira, Julio G. Segura, Beatriz A. Oliveira, Adja Cristina L. Medeiros, Rosa D. Zimba, Elisabete M.M. Viegas

PII:

S0377-8401(18)31540-2

DOI:

https://doi.org/10.1016/j.anifeedsci.2020.114393

Reference:

ANIFEE 114393

To appear in:

Animal Feed Science and Technology

Received Date:

3 December 2018

Revised Date:

31 October 2019

Accepted Date:

6 January 2020

Please cite this article as: Oliveira KRB, Segura JG, Oliveira BA, Medeiros ACL, Zimba RD, Viegas EMM, Distillers’ dried grains with soluble in diets for Pacu, Piaractus mesopotamicus juveniles: growth performance, feed utilization, economic viability, and phosphorus release, Animal Feed Science and Technology (2020), doi: https://doi.org/10.1016/j.anifeedsci.2020.114393

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Distillers' dried grains with soluble in diets for Pacu, Piaractus mesopotamicus juveniles: growth performance, feed utilization, economic viability, and phosphorus release Katia R. B. Oliveiraa1*, Julio G. Seguraa2, Beatriz A. Oliveiraa, Adja Cristina L. Medeirosa3, Rosa D. Zimbaa4, Elisabete M.M. Viegasa a

University of São Paulo, Ave. Duque de Caxias Norte, 225, Campus Fernando Costa, 13635-900,

Pirassununga, São Paulo - SP, Brazil

[email protected],

[email protected],

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[email protected],

[email protected], [email protected], [email protected]

*

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Corresponding author: Fax.: +55 35 3829 1250, E-mail address: [email protected]

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Present address 1

Federal University of Lavras - Ave. Doutor Sylvio Menicucci 1001, 37200-000, Lavras, MG,

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Brazil

Central University of Ecuador, Aquaculture Laboratory. Jerónimo Leiton, s/n y Gato Sobral –

2

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Quito – Ecuador. e-mail: [email protected] 3

Felipe Camarão St., 337, Ribeira, 59012-480, Natal, RN, Brazil

4

Escola Superior de Desenvolvimento Rural (ESUDER), 5° Congresso, Zona de Expansão -

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Vilankulo-Inhambane – Mozambique. E-mail: [email protected]

Highlights

1. Corn DDGS showed high protein and phosphorus digestibility for Piaractus mesopotamicus 2. Corn DDGS could totally replace soybean meal in diets for Piaractus mesopotamicus 3. Use of corn DDGS in fish diets reduced dietary costs and environmental impact.

Abstract Three experiments were conducted to evaluate corn distillers' dried grains with soluble (DDGS) digestibility and the effects of soybean meal replacement by corn DDGS in diets for P. mesopotamicus on growth performance, feed utilization, economic viability, and phosphorus release. In experiment 1, six replicates of fish (13 ± 0.3 g mean weight) were fed the referential and test diets to evaluate DDGS apparent digestibility for dry matter, protein, energy, lipids, and phosphorus. In experiment 2, four replicates of fish (21 ± 0.2 g mean weight) were fed

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experimental diets (0, 100, 200, 300, or 400 g/kg of DDGS inclusion) for 100 days. In experiment 3, five replicates of fish (27 ± 1.4 g mean weight) were fed the experimental diets to measure DDGS phosphorus release in the water. Corn DDGS presented a low apparent

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digestibility coefficient (ADC) for dry matter and energy, and high ADC for protein, lipids, and phosphorus. The feed conversion ratio and protein efficiency ratio were positively affected

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by DDGS inclusion. Dietary costs of weight gain were reduced by 26% with soybean meal

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replacement by DDGS. Total phosphorus release in the water decreased with increasing levels of DDGS in the diets. Thus, the results indicate that the inclusion of 400 g/kg of corn DDGS as a plant protein source in diets for P. mesopotamicus in total replacement of soybean meal

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improved fish production and reduced the environmental impact.

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Abbreviations: ADC, apparent digestibility coefficient; ADC rd, apparent digestibility coefficient of referential diet; ADC td, apparent digestibility coefficient of test diet; ADC ti, apparent digestibility coefficient of test ingredient; ADF, acid detergent fiber expressed inclusive of residual ash; aNDF, neutral detergent fiber assayed with a heat stable amylase and expressed inclusive of residual ash; BHT, butyl hydroxytoluene; CG, corn gluten; CM, corn meal; CY, carcass yield; DDGS, distiller’s dried grains with soluble; DDGS0, diet with 0g/kg of DDGS; DDGS100, diet with 100g/kg of DDGS; DDGS200, diet with 200g/kg of DDGS; DDGS300, diet with 300g/kg of DDGS; DDGS400, diet with 400g/kg of DDGS; DP, dissolved phosphorus; DP/DE, digestible protein/digestible energy ratio; D ref, referential diet; DS, distillers’ soluble; D test, test diet; ECR, economic conversion ratio; EDS, distillers’ soluble from corn endosperm; FBW, final body weight; FCR, feed conversion ratio; FI, feed intake; FM, fish meal; HPDDG, high protein distillers dried grain; HSI, hepatic somatic index; IBW, initial body weight; PER, protein efficiency ratio; PM, poultry meal; RB, rice bran; SBM,

soybean meal; SEM, standard error of the mean; SGR, specific growth rate; TDC, total dietary cost; TP, total phosphorus; VSI, visceral somatic index; WG, weight gain; WM, wheat meal.

Keywords: distiller´s grains; environmental impact; omnivorous; phosphorus availability; plant protein; residue

1. Introduction Residues are a challenge for industries since they have no more propose in the productive

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chain but, pose a problem for storage and disposal. The potential reuse of residues as ingredients in animal feed is an alternative that would benefit both industries and the environment. Corn distiller’s dried grain with soluble (DDGS) is the residue of grain

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fermentation by enzymes and yeasts in ethanol production (Wyman, 1996). Its chemical composition might vary according to the grain source and methods of fermentation (Lim et al.,

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2011; Liu, 2009), however, it basically consists of 268 to 337 g/kg of crude protein, 35 to 128

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g/kg of lipid, 250 to 513 g/kg of neutral detergent fiber, 175 to 246 g/kg of acid detergent fiber, 28 to 98 g/kg of ash, 5 to 11 g/kg of lysine, and 5 to 8 g/kg of methionine (Ayadi et al., 2012). In aquaculture, the use of corn DDGS has been studied since the 1980s (Lovell, 1980).

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Recent studies have reaffirmed the potential of DDGS in diets for several species with importance in aquaculture, such as channel catfish, Ictalurus punctatus (Li et al., 2011), Nile

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tilapia, Oreochromis niloticus (Suprayudi et al., 2015), and rainbow trout, Oncorhynchus

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mykiss (Overland et al., 2013; Welker et al., 2014). In addition, some researchers have focused on DDGS digestibility for Pacific white shrimp, Liponeaus vannamei (Lemos et al., 2009), channel catfish (Li et al., 2011), meagre, Argyrosomus regius, European seabass, Dicentrarchus labrax (Magalhães et al., 2015), and sunshine bass, Morone chrysops x Menticirrhus saxatilis (Thompson et al., 2008), showing the potential use of corn DDGS as a plant protein source in the diets of a variety of aquatic species.

However, corn DDGS might be an efficient source of protein mainly for omnivorous species, since their requirements for protein are not as marked as those of carnivorous food species (Boscolo et al., 2011; Gatlin et al., 2007; Hardy, 2010). In addition, omnivorous fish can tolerate up to 90 g/kg of crude fiber in the diet (Rodrigues et al., 2010), which minimizes the limitations of using DDGS due to its high fiber content. Commonly known as Pacu or Pacu-caranha, Piaractus mesopotamicus is a tropical fish species common in Latin America. It is omnivorous, feeding on soybean meal as the major

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source of plant protein in commercial diets. Pacu has great potential for production in captivity due to its rusticity, good environmental adaptability, and high growth rates (Jomori et al., 2005), which potentiate this species for aquaculture production in tropical areas.

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Thus, the aim of this study was to evaluate corn DDGS digestibility and the effects of its inclusion at different levels replacing soybean meal in diets for juvenile P. mesopotamicus on

2. Material and methods

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growth performance, feed utilization, economic viability, and phosphorus release.

The use of animals this research was approved by the Ethics Committee of Animal Welfare

number 1910/16.

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of the College of Agricultural and Veterinary Sciences of Sao Paulo State University, protocol

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Three trials, experiment 1, experiment 2, and experiment 3, evaluated corn DDGS

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digestibility and the effects of its inclusion as a dietary replacement of soybean meal in diets for Pacu, P. mesopotamicus. Experiment 1 evaluated the apparent nutrient digestibility of corn DDGS. Experiment 2 evaluated the effect of feeding diets with increasing DDGS levels on fish growth performance, feed utilization, and economic viability, and experiment 3 evaluated the total and dissolved phosphorus release in the culture water with varying levels of DDGS inclusion in the diet.

2.1. Experimental diets Two diets were formulated to determinate the apparent nutrient digestibility of corn DDGS: a reference diet containing 320 g/kg of crude protein, 17.6 MJ/kg of gross energy, and 5 g/kg of chromic oxide III (Cr2O3); and a test diet consisting of 70% of the reference diet and 30% of corn DDGS. Corn DDGS was supplied by Libra Etanol LTDA, Sao Jose do Rio Claro, Mato Grosso, Brazil. The composition and approximate analysis of the ingredients and experimental diets are shown in Table 1 and Table 2, respectively.

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After obtaining the apparent digestibility coefficients of DDGS nutrients, five diets with increasing levels of DDGS inclusion (0, 100, 200, 300, and 400 g/kg) were formulated to be, on a digestible basis, isoproteic (290 g/kg digestible protein) and isoenergetic (13.4 MJ/kg

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digestible energy). The apparent digestibility coefficients of soybean meal, fishmeal, wheat meal, corn, rice bran, poultry meal (Abimorad and Carneiro, 2004), and corn gluten meal

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(Fabregat et al., 2008) previously obtained for P. mesopotamicus were used to formulate the

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experimental diets. The approximate composition of experimental diets is shown in Table 3. The dietary ingredients were ground to 1 mm and mixed in a single screw extruder (Imbramarq, model Labor PQ-30). The extrusion temperature was kept at 124°C, screw speed

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at 676.5 rpm, feeding rate at approximately 500 g/min, and the circular matrix was set at 2 mm. Pellets were dried at 50°C for 12 hours and then stored in a freezer (-20°C) until use.

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2.2. Fish and culture conditions

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Pacu (P. mesopotamicus) juveniles were obtained from a commercial fish farm and acclimatized for 30 days upon arrival to the experimental facilities. During this period, fish were fed a commercial diet (Pira 32, Guabi, 320 g/kg of crude protein) twice daily. 2.2.1. Experiment 1 The apparent digestibility coefficients of DDGS were measured by the indirect method, following the methodology described by the NRC (2011). For that, 210 Pacu juveniles (13.6 ±

0.3 g mean weight) were randomly distributed in six rectangular tanks (50 L) at a density of 35 fish/tank. The tanks were maintained in a recirculated system supplied with aeration and the temperature was kept constant by heaters (26°C). Each tank was considered as an experimental unit, arranged in a completely randomized design with two treatments (reference and test diet) and six replicates. The water quality was maintained by the use of supplemental aeration (central line and air diffusers), and mechanical and biological filtration. The collection of feces was divided into two periods, one for each treatment (n = 6

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replicates per diet). In each period, fish were fed with the respective diet for seven days for adaptation. During diet adaptation, fish were fed to apparent satiation twice daily (09:00 and 17:00 hours). Feces were collected using a Guelph modified system as described by Abimorad

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and Carneiro (2004), with modifications. One hour after the last daily feeding, the fish were transferred to six conic fiber glass (100 L capacity) collection tanks with a valve connecting

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the bottom of the tank to a Falcon tube kept on ice. The collection tanks were maintained in a

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closed water flow system, supplied with aeration and heaters to maintain the water temperature at 26°C. Ten hours later, feces settled on the Falcon tubes were collected and the fish were relocated back into their initial tanks (50 L). The procedure was repeated for four days to obtain

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enough feces for future analyses. The same procedure was used in the two collection periods (for the reference and test diets). After collection, the feces were centrifuged (1800 × g, 10

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min), stored at -20°C, and later lyophilized prior to chemical analysis.

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The apparent digestibly coefficients (ADCs) of the dry matter, protein, energy, lipid, and phosphorus of the experimental diets were calculated according to NRC (2011) as follows: ADC = 100*[1 – (Cr2O3 diet/Cr2O3 feces)*(Nutrient or Energy feces/Nutrient or Energy diet)]. The apparent digestibility coefficients of protein, dry matter, energy, lipid and phosphorus of the test ingredient (DDGS) were calculated according to NRC (2011): ADC ti = ADC td + [(ADC td - ADC rd)*(0.7*D ref/0.3*D test ingredient)], where ti = test ingredient; td = test

diet; rd = reference diet; D ref is the % nutrient (or MJ/kg) of reference diet (dry matter basis) and D test ingredient is the % nutrient (or MJ/kg) of test ingredient (dry matter basis). 2.2.2. Experiment 2 To set up the trial, a total of 300 fish were weighed (mean weight 21 ± 0.2 g) and distributed in 20 fiber glass tanks (100 L) at density of 15 fish/tank. Each tank was considered as an experimental unit, arranged in a completely randomized design with five treatments: inclusion of 0, 100, 200, 300, and 400 g/kg of DDGS (DDGS0, DDGS100, DDGS200, DDGS300, and

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DDGS400, respectively) and four replicates. The water quality was maintained using supplemental aeration (central line and air diffusers), and mechanical and biological filtration. The water temperature was controlled with

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a heat exchanger (26°C) and measured twice daily. The dissolved oxygen and pH were monitored daily using a multiparameter (U-10, Horiba, Kyoto, Japan). Total ammonia and

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nitrite were measured using commercial kits (Labcon Test Fresh Water Toxic Ammonia and

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Labcon Test Nitrite NO2-). The fish were fed with experimental diets for 100 days, twice daily (9:00 and 17:00), until apparent satiation. 2.2.3. Experiment 3

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To determinate the phosphorus release in the water, 150 Pacu juveniles (27.09 ± 1.4 g mean weight) were distributed in five fiber glass tanks (100 L) at a density of 30 fish/tank.

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Each tank was considered an experimental unit and the experiment was laid out in a 5 x 5 Latin

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square design (five treatments and five periods). The fish were fed for seven days, twice daily (09:00 and 17:00 hours), with the diets with

increasing DDGS levels (DDGS0, DDGS100, DDGS200, DDGS300, and DDGS400) to apparent satiation. One hour after the last feeding, the fish were transferred to five conic tanks (100 L) for water collection. Immediately before transferring the fish and exactly 10 hours later, a sample of 200 mL of water was taken to analyses the dissolved and total phosphorus

released in the water by the fish excretions. Afterwards, the fish were returned to their initial tanks, thus ending one period of the trial. The water samples were stored in the freezer at -20°C until analysis. The amount of dissolved and total phosphorus released in the water was determined by the difference between the samples collected at the initial time point and the samples collected at the end of the period. 2.3. Sampling At the end of experiment 2, all the fish were fasted for 24 hours, anaesthetized in

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benzocaine (50 mg/L), and slaughtered by spinal cord section. Then, each fish was weighed and gutted to obtaining the following parameters: Weight Gain = 100*(Final Body Weight (FBW) - Initial Body Weight (IBW))/IBW; Feed Intake (FI) = Total Feed Intake (g dry matter);

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Feed Conversion Ratio (FCR) = Dry Feed Intake/Wet Weight Gain; Specific Growth Rate (SGR) = [(log FBW – log IBW) / Period (days)]*100; Protein Efficiency Ratio (PER) = Wet

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Weight Gain/Crude Protein Intake; Carcass Yield (CY) = (Carcass Weight/FBW)*100; Viscera

Weight/FBW)*100. 2.4. Chemical analyses

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Somatic Index (VSI) = (Viscera Weight/FBW)*100; Hepatic Somatic Index (HSI) = (Liver

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Chemical analyses of the ingredients and diets were determined according to the Association of Official Agricultural Chemists (AOAC, 2012) methodology. The ingredients

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and diets were plated in a forced air-drying oven (105°C) for dry matter determination (930.15).

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The samples were analyzed for their crude protein content (N*6.25) according to the Kjeldahl method (968.06), for their ash content (942.05) by incineration in a muffle oven (450°C for 16 h), and for their crude lipid content by extraction with petroleum ether using a Soxhlet system (920.39). The neutral detergent fiber (aNDF) and acid detergent fiber (ADF) content of the DDGS and diets (DDGS0, DDGS100, DDGS200, DDGS300, and DDGS400) were determined using the sequential method with the Ankom® Filter Bag Technique and heat stable α-amylase

(973.18). The starch content was determined colorimetrically using an ethanol extraction technique, followed by α-amylase and amyloglucosidase preparation to hydrolyze starch to glucose, as described by Hendrix (1993). Data for the aNDF, ADF, and starch content of the other ingredients were obtained from INRA/CIRAD/AFZ (2018). The gross energy was estimated using the Atwater general factor system (FAO, 2012; NRC, 2011) where gross energy (MJ/Kg) was defined as the sum of the total protein, lipid, and carbohydrate multiplied by their respective conversion factor, based on the heat generated after combustion: 4.0 kcal/g

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for protein, 9.0 kcal/g for lipids, and 4.0 kcal/g for carbohydrates. The phosphorus content of DDGS was determined by absorbance in a spectrophotometer (882 nm) after acid digestion using the molybdenum colorimetric method (Silva and Queiroz, 2002). The determination of

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the total and dissolved phosphorus concentration in the water was based on the formation of the antimony-phospho-molybdate complex and its reduction to an intensely blue-colored

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complex by ascorbic acid (Mackreth et al., 1978). Dissolved phosphorus was determinate after

2.5. Economic viability

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filtration of the sample (Mackreth et al., 1978).

The economic viability of the use of corn DDGS in diets with increasing DDGS levels

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(DDGS0, DDGS100, DDGS200, DDGS300, and DDGS400) for Pacu juveniles was assessed following the methodology described by Hoffman (2006) and the suggestions by Gameiro

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(2009). Data on the costs of the dietary ingredients was collected for the last 10 years (2005 to

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2015) from the Institute of Agricultural Economics (IEA/APTA, Brazil) to determinate the total dietary cost (TDC) and economic conversion ratio (ECR) of the experimental diets, using the following equations: TDC (US$/kg) = (% ingredient in diet*ingredient cost)/100) and ECR (US$/kg) = TDC*FCR. The currency used for the economic evaluation was the US dollar (US$), based on data from the Brazilian Central Bank (14 April 2016). 2.6. Statistical analyses

The data are expressed as mean ± pooled standard error of the mean (SEM). The normality and homogeneity of variances were tested using the Shapiro-Wilk and Levene tests, respectively. All statistical procedures were performed using the SPSS software package (SPSS, 2013). Data were analyzed by one-way ANOVA and in case of significance, an orthogonal polynomial contrast was applied to identify linear and quadratic effects of dietary DDGS inclusion.

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3. Results The nutrient digestibility of the experimental diets with increasing DDGS inclusion was not the subject of the present study, therefore, the data are presented elsewhere. In short, the

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ADC of dry matter and energy decreased with dietary DDGS inclusion, whereas the ADC of the lipids increased, and no differences were observed for protein digestibility.

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3.1. Experiment 1

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The apparent digestibility coefficients for the dry matter, protein, energy, lipids, and phosphorus of corn DDGS are shown in Table 4. A higher digestibility of corn DDGS protein,

3.2. Experiment 2

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phosphorus, and lipids was observed than of the energy and dry matter.

The growth parameters and feed efficiencies are presented in Table 5. Increasing dietary

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levels of DDGS improved the FCR and PER. No differences were observed for weight gain,

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feed intake, specific growth rate, the visceral somatic and hepatic somatic indexes, or carcass yield. The FCR decreased linearly from 1.14 for DDGS0 to 1.03 for the DDGS400 diet. On the contrary, the PER increased linearly from DDGS0 to DDGS400. Data on the economic viability are presented in Table 6; corn DDGS inclusion reduced the total dietary cost and the economic conversion ratio. 3.3. Experiment 3

The data for dissolved and total phosphorus released in the water during the experiment are shown in Table 7. The amount of total phosphorus released in the water varied with the treatments. However, the same was not true for dissolved phosphorus, which was unaffected by the treatments. The total phosphorus released in the water decreased linearly with increasing dietary DDGS inclusion, ranging from 0.61 mg/L for DDGS0 to 0.37 mg/L for the DDGS400 diet.

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4. Discussion 4.1. Experiment 1

The success of fish rearing mostly depends on the quality of food supplied in fish farms.

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Thus, exhibiting the nutrient availability of dietary ingredients is important practice to allow the maximum expression of animal productive performance and to reduce nutrient waste

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through fish excretions (Lee et al., 2002). The availability of nutrients can be calculated by

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determination of the ingredients’ apparent digestibility coefficients (NRC, 2011; Smith et al., 1995), which is recognized as the first step in evaluating the potential use of ingredients in animal feed production (Allan et al., 2000). Thus, corn DDGS apparent digestibility

present study.

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coefficients (ADC) were first determined for Pacu (Piaractus mesopotamicus) juveniles in the

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Through the dry matter digestibility, it is possible to get a general estimate of the

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digestibility of a particular ingredient by indicating the amount of non-digestible nutrients present in it. As calculated for Pacu, the low dry matter digestibility of corn DDGS has been reported for the majority of the aquatic species studied to date (Chan et al., 2004; Li et al., 2011; Magalhães et al., 2015; Seo et al., 2011). Among the factors capable to induce changes in the digestibility of dietary nutrients, fiber is the most significant when talking about DDGS. The inclusion of large amounts of fiber in diets would lead to a reduction in gut retention time

of digesta and in the contact of digestive enzymes with intracellular contents (Vanderhoof, 1998), which impairs the animal’s ability to absorb nutrients available from dietary ingredients (Enes et al., 2011; Fontoulaki et al., 2005; Stone et al., 2003). Additionally, a high fiber content can lead to an increase in microbial activity and substrates from fermentation, which would result in endogenous losses. The ADC of protein obtained for DDGS was higher than that obtained for other ingredients most often used as a protein source in Pacu diets, such as poultry meal (0.83), soybean meal

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(0.81) (Abimorad and Carneiro, 2004), corn gluten meal (0.79) (Fabregat et al., 2008), fish meal (0.85), and yeast extract (0.82) (Abimorad et al., 2008). In addition, the ADC of protein in DDGS for Pacu was higher than the 0.86 reported for channel catfish (Li et al., 2011), or the

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0.64 reported for sunshine bass (Thompson et al., 2008). Thus, this highlights the potential use of DDGS as a protein source in Pacu diets. However, even though DDGS has shown a good

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digestible protein value, more detailed studies of DDGS’ amino acid profile and digestibility

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for Pacu are required. Despite their different food habits, when studying meagre and seabass, Magalhães et al. (2015) reported a lower digestibility of amino acids in diets with DDGS inclusion than in fishmeal-based diets, probably due to the lower availability of amino acids in

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DDGS than in fishmeal.

All the energy released by DDGS comes, basically, from lipids, since starch is degraded

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in the fermentation process of DDGS production. Nevertheless, in the present study, the source

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of corn DDGS incorporated in the diets had lipids level that were not as high as those reported in the literature. That might be a result of the extraction of the oil retained in the wet cake during ethanol production, which generates corn oil as another ethanol co-product. Therefore, variations in the lipid content of DDGS will be dependent on the process adopted by ethanol plants in DDGS production. In addition, the starch that remained in the DDGS used in this experiment did not have a remarkable contribution as an energy source. Hence, the low value

of the apparent digestibility coefficient obtained for the energy in DDGS for Pacu might be explained by the quantity of nutrients with low energy digestibility, such as fibre, and low amounts of nutrients with higher digestible values, such as starch and lipid (Stein and Bohlke, 2007; NRC, 2011). The digestibility of the lipids present in DDGS was lower than that reported for other common ingredients used in Pacu diets such as corn (0.91), soybean meal (0.93), and fish meal (0.94) (Furuya et al., 2001). In addition, DDGS had a lower lipid digestibility for Pacu than for

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channel catfish (0.94) (Li et al., 2011), but higher than the 0.69 reported for sunshine bass (Thompson et al., 2008). This might be due to differences in the lipid content of the diets, which was 40 g/kg in the present study, 130 g/kg for the sunshine bass study, and 70 g/kg for the

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channel catfish study.

The apparent phosphorus digestibility of DDGS was considerably higher than that of

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ingredients such as soybean meal (0.35), fishmeal (0.27), bone meal (0.55), corn meal (0.07),

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and wheat bran (0.31) for Nile tilapia fingerlings (Miranda et al., 2000). Furthermore, the digestibility of phosphorus in DDGS for Pacu juveniles was higher than that observed for channel catfish (Li et al., 2015) and pigs (Almeida et al., 2012; Rojas et al., 2013). Digestible

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phosphorus refers to the portion absorbed by the gastrointestinal tract. This portion can be compounded by non-phytic or phytic phosphorus, which is the last product available after the

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hydrolysis process by intrinsic phytase (Bünzen, 2008; Cao et al., 2007). Thus, the high

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digestible phosphorus content found in DDGS might be explained by its high bioavailability. The fermentation processes and heating that DDGS is subjected to might have resulted in an increase in the hydrolysis of phytate molecules (Kim et al., 2008), which are non-digestible for animals. It is important to emphasize that variations in the quality and nutrient composition of the DDGS among different facilities or even in the same facility might occur (Kleinschmitt et al.,

2007; Spiehs et al., 2002) due to differences between plants and their process methods (Liu, 2011; Ortin and Yu, 2009). Therefore, the success of DDGS inclusion in fish diets as well as the efficiency of nutrient utilization by the fish will be dependent the source and composition of the DDGS being used. 4.2. Experiment 2 In agreement with the present study, a decrease in the FCR was observed for rainbow trout (143 g mean initial weight) fed diets with 500 g/kg of DDGS as a substitute for a mixture with

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plant protein ingredients (Overland et al., 2013), without significant effects on weight gain and feed intake. Robinson and Li (2008) observed that the lower FCR found for channel catfish fed 300 g/kg DDGS in the diet was not due to improvements in feed intake but to an increase in

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fish weight gain. Improvements in feed utilization were reported for channel catfish fed diets with DDGS, DS (distillers’ soluble), and EDS (distillers’ soluble from corn endosperm)

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compared to HPDDG (high protein distillers dried grain), probably due to the absence of the

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soluble portion in HPDDG (Li et al., 2010). Thus, it is possible to confirm that some components of the soluble portion of DDGS might have facilitated feed utilization by the fish. Some studies on the inclusion of brewer’s yeast in fish diets have shown improvements in

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feed efficiency (Li and Gatlin, 2006; Oliva-Teles and Gonçalves, 2001). The benefits of yeast inclusion in Pacu diets include improvements in feed efficiency, growth performance,

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digestibility of nutrients and amino acids, and a reduction in nitrogen release (Ozorio et al.,

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2010). The nucleotides, β glucans, and oligosaccharides present in yeast cells have been reported to modify the intestine morphometry of some fish species, increasing the area for nutrients absorption (Burrells et al., 2001; Dimitroglou et al., 2009; Santin et al., 2001; Yang et al., 2007) and consequently the use of the dietary nutrients. Since DDGS is composed of considerable amounts of yeast cells (Ingledew, 1999; Zohu et al. 2010), it is possible that its components modified the Pacu intestine, improving the absorption of dietary nutrients, and

exercising a positive influence on Pacu growth performance, feed utilization, and nutrient digestibility. The improvements in feed utilization observed for Pacu juveniles fed with DDGS diets in the present study were reflected in the economic viability. Total replacement of soybean meal by corn DDGS promotes a reduction of 17% of the total dietary cost and 26% of the economic conversion ratio. Once feed contributes 60 to 80% of the total production costs (Rola and Hasan, 2007), the use of low-cost ingredients in aqua feed becomes priority, as long as it does

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not cause losses in animal production. 4.3. Experiment 3

An increase in nutrient retention leads to a smaller quantity of nutrients being released in

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water as waste (Bureau and Hua, 2010; Prachom et al., 2013). In the present study, the discharge of phosphorus in the water was evaluated. The results obtained are in agreement with

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Cheng and Hardy (2004) and Overland et al. (2013), who found that diets with DDGS inclusion

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promoted better phosphorus retention for rainbow trout. Conama 357/2005 recommends values above 0.05 mg/L of phosphorus for aquaculture water, therefore, only the experimental diets with 300 and 400 g/kg of DDGS inclusion were in the phosphorus concentration range

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recommended. Although corn DDGS has a higher phosphorus content availability than other ingredients commonly used in diets for aquaculture, its use is recommended as a strategy for

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the development of complete and appropriate aqua feed since it allows better phosphorus

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utilization by the fish and reduces the amount of phosphorus excreted into the environment.

5. Conclusion

This is the first work to investigate corn distillers' dried grains with soluble digestibility and its inclusion in total replacement of soybean meal in diets for Piaractus mesopotamicus. The results indicate that the inclusion of 400 g/kg of corn distillers' dried grains with soluble

as a plant protein source in diets for Piaractus mesopotamicus in total replacement of soybean meal improved fish production and reduced the environmental impact.

Conflict of interest The authors declare that they have no competing interest.

Acknowledgments

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This work was supported by the National Council for Technological and Scientific Development (CNPq) (Process 130664/2014-6) and São Paulo Research Foundation

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(FAPESP) (Process 2014/16685-5).

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Table 1 Proximate analyses of experimental dietary ingredients (dry matter basis) 1

1

SBM

CM

WM

RB

CG

FM

PM

929 336 20.9 564 286 40 20 2 53

889 448 17.1 148 88 20 78 6 -

893 84 16.2 125 31 40 13 3 -

903 148 16.7 458 136 29 55 9 -

869 132 19.4 229 102 152 102 16 -

927 715 21.1 40 14 19 22 4 -

977 580 14.6 34 275 30 -

973 489 13.1 152 150 27 -

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Chemical Composition (g/kg) Dry matter (g/kg as fed) Crude protein Gross energy (MJ/kg) aNDF ADF Lipid Ash Phosphorus Starch

DDGS

Ingredients referenced to diets of experiments 1, 2 and 3. DDGS, corn distiller’s dried grains

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with soluble (Libra Etanol, Mato Grosso, Brazil); SBM, soybean meal; CM, corn meal; WM, wheat meal; RB, rice bran (Cargill, Sao Paulo, Brazil); CG, corn gluten; FM, fish meal (In

detergent fiber; ADF, acid detergent fiber.

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Table 2

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Vivo, Sao Paulo, Brazil); PM, poultry meal (Agromix, Sao Paulo, Brazil); aNDF, neutral

Formulation and chemical composition of reference and test diets1

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Reference Diet

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Ingredients (g/kg) DDGS Poultry meal 145 Fishmeal 60 Soybean meal 300 Wheat meal 240 Corn 150 Rice bran 90 BHT 1 Chromium oxide III 5 2 Vit. And Min. Premix 10 Total 1000 Chemical Composition (g/kg dry matter) Dry matter (g/kg as fed) 935 Crude protein 362

Test Diet 300 101 42 209 167 105 63 1 5 7 1000 945 355

Gross energy (MJ/kg) Crude lipid Phosphorus aNDF* ADF*

18.8 34 10 194 73

19.2 45 8 304 137

1

Diets referenced to experiment 1.

2

Vitamin and Mineral Premix (g/kg of diet): vitamin A - 5.000 UI; vitamin D3 – 2.500 UI;

vitamin E - 50 mg; vitamin K3 - 5 mg; vitamin B1 - 15 mg; vitamin B2 - 15 mg; vitamin B6 15 mg; vitamin B12 - 40 mg; folic acid - 5 mg; pantothenate Ca - 40 mg; vitamin C - 100 mg;

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biotin – 0.1 mg; Inositol - 10; nicotinamide - 70; choline - 100 mg; Co – 0.1 mg; Cu - 10 mg; Fe - 50 mg; I - 2 mg; Mn - 15 mg; Se – 0.3 mg; Zn – 90 mg. (Agromix LTDA, Sao Paulo, Brazil) *

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Values estimated based on dietary ingredients chemical composition (INRA/CIRAD/AFZ,

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2018) and laboratorial analysis for DDGS.

BHT, butyl hydroxytoluene; DDGS, corn distiller’s dried grains with soluble; aNDF, neutral

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detergent fiber; ADF, acid detergent fiber. Table 3

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Formulation and chemical composition of the experimental diets1 with increasing levels of

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DDGS inclusion (dry matter basis)

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Ingredients (g/kg) DDGS Soybean meal Corn Soybean oil Poultry meal Fish meal Corn Gluten Wheat meal Rice bran Lysine2 BHT

DDGS0

DDGS100

DDGS200

DDGS300

DDGS400

0 231 254 34 66 110 83 130 77 5 1

100 173 218 28 66 110 83 130 77 5 1

200 115 181 23 66 110 83 130 77 5 1

300 58 145 17 66 110 83 130 77 5 1

400 0 108 11 66 110 83 130 77 5 1

1

10 1000

10 1000

10 1000

10 1000

916 321 19.1 52 78 281 188 196 276 14.7 18.8

934 327 19.4 58 69 328 200 181 294 14.9 19.7

913 321 19.6 71 69 387 223 157 282 12.5 22.6

938 327 19.9 76 66 442 221 143 291 14.5 20.1

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Vit. and Min. Premix3 10 Total 1000 Chemical Composition (g/kg dry matter) Dry matter (g/kg as fed) 931 Crude protein 328 * Gross energy (MJ/kg) 18.6 Crude lipid 32 Ash 82 aNDF 249 ADF 138 Starch 228 Digestible protein 295 Digestible energy (MJ/kg) 14.7 DP/DE (g/MJ) 20.1

Diets with inclusion of 0, 100, 200, 300 and 400 g kg-1 of corn DDGS (DDGS0, DDGS100,

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DDGS200, DDGS300, DDGS400) referenced to experiments 2 and 3. Lysine: Anjinomoto LTDA, Sao Paulo, Brazil

3

Vitamin and Mineral Premix (g/kg of diet): vitamin A - 5.000 UI; vitamin D3 – 2.500 UI;

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2

vitamin E - 50 mg; vitamin K3 - 5 mg; vitamin B1 - 15 mg; vitamin B2 - 15 mg; vitamin B6 -

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15 mg; vitamin B12 - 40 mg; folic acid - 5 mg; pantothenate Ca - 40 mg; vitamin C - 100 mg; biotin – 0.1 mg; Inositol - 10; nicotinamide - 70; choline - 100 mg; Co – 0.1 mg; Cu - 10 mg;

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Fe - 50 mg; I - 2 mg; Mn - 15 mg; Se – 0.3 mg; Zn – 90 mg. (Agromix LTDA, Sao Paulo, Brazil)

Values calculated according to FAO (2012) and NRC (2011)

ur

*

BHT, butyl hydroxytoluene; DDGS, corn distiller’s dried grains with soluble; aNDF, neutral

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detergent fiber; ADF, acid detergent fiber; DP/DE, digestible protein /digestible energy ratio.

Table 4 Apparent digestibility coefficients of corn DDGS for P. mesopotamicus juveniles Test Ingredient Corn DDGS 0.61 0.95 0.65 0.89 0.98

Item Dry matter Protein Energy Lipids Phosphorus 1

SEM 0.055 0.009 0.046 0.027 0.049

Values referenced to experiment 1, presented as means (six replicates per treatment and 35

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fish per replicate) and pooled standard error of the mean (SEM). DDGS, distiller’s dried grains

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with soluble.

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Table 5

1

DDGS300 20.9 113.2 431.3 92.4 1.00 1.7 3.2 85.9 9.3 1.4

-p

DDGS200 20.6 120.3 492.4 107.2 1.07 1.8 2.8 93.3 10.3 1.4

re

DDGS100 21.1 100.1 412.3 88.2 1.04 1.6 3.0 91.5 8.4 1.5

lP

IBW (g) FBW (g) WG (%) FI (g % DM) FCR SGR PER CY (%) VSI (%) HSI (%)

Treatments* DDGS0 21.5 101.1 365.1 90.5 1.14 1.5 2.7 90.3 8.8 1.6

DDGS400 20.7 115.1 471.7 97.1 1.03 1.7 3.0 87.5 9.2 1.4

SEM 0.13 3.52 22.14 10.71 0.027 0.09 0.07 2.51 0.89 0.11

Contrast Linear 0.067 0.266 0.243 0.614 0.011 0.149 0.007 0.184 0.573 0.288

Quadratic 0.206 0.704 0.650 0.625 0.170 0.340 0.084 0.386 0.487 0.465

rn a

Item

ro

Growth parameters and feed utilization efficiency of P. mesopotamicus juveniles fed experimental diets1

Values referenced to experiment 2, presented as means (four replicates per treatment and 15 fish per replicate) and pooled standard error of the

mean (SEM). *

Jo u

Diets with inclusion of 0, 100, 200, 300 and 400 g/kg of corn DDGS (DDGS0, DDGS100, DDGS200, DDGS300, DDGS400).

One-way ANOVA was applied; among groups orthogonal polynomial contrast was applied to identify linear and quadratic effects. IBW, initial body weight; FBW, final body weight; WG, weight gain; FI, feed intake in dry matter basis; FCR, feed conversion ratio; SGR, specific growth rate; PER, protein efficiency ratio; CY, carcass yield; VSI, visceral somatic index; HIS, hepatic somatic index; DDGS, distiller’s dried grains with soluble.

Table 6 Dietary costs1 of diets with increasing levels of corn DDGS in soybean meal replacement for P. mesopotamicus juveniles

Item FCR TDC (US$/kg) ECR (US$/kg)

DDGS100 1.04 0.53 0.55

DDGS200 1.07 0.51 0.54

DDGS300 1.00 0.49 0.49

DDGS400 1.03 0.46 0.47

Costs of ingredients based on IEA (2016).

ro of

1

Treatments* DDGS0 1.14 0.56 0.64

*

Diets with inclusion of 0, 100, 200, 300 and 400 g/kg of corn DDGS (DDGS0, DDGS100,

DDGS200, DDGS300, DDGS400).

FCR, feed conversion ratio; TDC, total dietary cost; ECR, economic conversion ratio; DDGS,

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ur

na

lP

re

-p

distiller’s dried grains with soluble.

of

Table 7

DP (mg/L) TP (mg/L)

DDGS100 0.04 0.06

DDGS200 0.03 0.05

DDGS300 0.03 0.04

DDGS400 0.04 0.05

SEM 0.002 0.003

Contrast Linear Quadratic 0.351 0.196 0.001 0.421

Values referenced to experiment 3, presented as means (n = 25, five tanks and five periods) and pooled standard error of the mean (SEM).

re

1

Treatments* DDGS0 0.04 0.06

-p

Item

ro

Total phosphorus and dissolved phosphorus release in water of P. mesopotamicus fed diets with increasing levels of DDGS1.

*

lP

Diets with inclusion of 0, 100, 200, 300 and 400 g/kg of corn DDGS (DDGS0, DDGS100, DDGS200, DDGS300, DDGS400).

One-way ANOVA was applied; among groups orthogonal polynomial contrast was applied to identify linear and quadratic effects.

Jo u

rn a

DP, dissolved phosphorus; TP, total phosphorus; DDGS, distiller’s dried grains with soluble.