950 V.L. da Cunha et al. Feeding rate and frequency on juvenile pompano growth Viviana Lisboa da Cunha(1), Marcelo Roberto Pereira Shei(1), Marcelo Hideo Okamoto(1), Ricardo Vieira Rodrigues(1) and Luís André Sampaio(1) Universidade Federal do Rio Grande, Instituto de Oceanograia, Laboratório de Piscicultura Estuarina e Marinha, Caixa Postal 474, CEP 96201‑900 Rio Grande, RS, Brazil. E‑mail: viviana_aquicultura@yahoo.com.br, marceloshei@gmail.com, mar_okamoto@yahoo.com.br, vr.ricardo@gmail.com, sampaio@mikrus.com.br (1) Abstract – The objective of this work was to evaluate the ideal feeding rate and frequency for juvenile pompano (Trachinotus marginatus). Two experiments were carried out in a completely randomized design, with three replicates each. In experiment I, 25 ish (4.8±0.6 g and 6.48±0.01 cm) were stocked in 15 tanks (50 L) during 21 days and fed 4, 8, 12, 16, and 20% body weight per day. In experiment II, 20 ish (4.1±0.1 g and 6.6±0.1 cm) were stocked in 15 tanks (40 L) during 28 days and fed 2, 6, 8, and 10 times a day. The tested feeding rates and frequencies did not inluence survival. Final weight and length in experiment I were signiicantly lower in ish fed 4% body weight per day, whereas in experiment II only weight was signiicantly lower in ish fed 2 and 6 times a day. At the end of both experiments, apparent feed conversion showed signiicant difference, with the worst value observed for ish fed 20% body weight per day in experiment I and 2 times a day in experiment II. Juvenile pompano show better growth performance when fed 8% body weight per day and 8 times a day. Index terms: Trachinotus marginatus, feed management, marine ish culture. Taxa e frequência alimentar no crescimento de juvenis de pampo Resumo – O objetivo deste trabalho foi avaliar a taxa e a frequência alimentar ideal para juvenis de pampo (Trachinotus marginatus). Foram conduzidos dois experimentos, em delineamento inteiramente casualizado, com três repetições cada um. No experimento I, 25 peixes (4,8±0,6 g e 6,48±0,01 cm) foram estocados em 15 tanques (50 L) durante 21 dias e alimentados com 4, 8, 12, 16 e 20% da biomassa total por dia. No experimento II, 20 peixes (4,1±0,1 g e 6,6±0,1 cm) foram estocados em 15 tanques (40 L) durante 28 dias e alimentados 2, 6, 8 e 10 vezes por dia. As taxas e as frequências alimentares testadas não inluenciaram a sobrevivência. O peso e o comprimento inais no experimento I foram signiicativamente menores nos peixes alimentados com 4% da biomassa total por dia, enquanto, no experimento II, somente o peso foi signiicativamente menor nos peixes alimentados 2 e 6 vezes por dia. Ao inal dos dois experimentos, a conversão alimentar aparente apresentou diferença signiicativa, com pior valor observado nos peixes alimentados com 20% da biomassa total por dia, no experimento I, e 2 vezes por dia no experimento II. Juvenis de pampo apresentam melhor desempenho de crescimento quando alimentados com 8% da biomassa total por dia e oito vezes por dia. Termos para indexação: Trachinotus marginatus, manejo alimentar, piscicultura marinha. Introduction The selection of new ish species with good potential for aquaculture is crucial for the development of this growing industry (Tutman et al., 2004). Carangidae ish support a diverse array of economically important isheries in tropical and subtropical waters worldwide (Crabtree et al., 2002), and species of Trachinotus have already been considered suitable candidates for mariculture due to their easy adaptation to culture systems, acceptance of formulated feeds, and fast growth rates (Crabtree et al., 2002; Tutman et al., 2004; Weirich et al., 2006). Pesq. agropec. bras., Brasília, v.48, n.8, p.950-954, ago. 2013 DOI: 10.1590/S0100-204X2013000800020 One of the species that stands out is the pompano (Trachinotus marginatus), an euryhaline ish (Sampaio et al., 2003) that inhabits shallow coastal waters from Rio de Janeiro, Brazil, to Uruguay (Menezes & Figueiredo, 1980). Several researches have been conducted to assess the culture of this species (Sampaio et al., 2003; Chaves et al., 2006; Cunha et al., 2009; Okamoto et al., 2009; Kütter et al., 2012, 2013). However, in order to evaluate and develop ideal conditions for the production of new aquaculture species, among others requirements, it is necessary to establish an appropriate feed protocol. The determination of feed protocol is important to improve Feeding rate and frequency on juvenile pompano growth environmental safety and economic return (Cho et al., 2007; Kim et al., 2007), and one of the ways to achieve this goal is by determining ish feeding rate and frequency. Studies have shown that a suitable feeding rate and frequency improve growth and feed conversion (Wang et al., 1998; Van Ham et al., 2003; Cho et al., 2007), reduce variation in ish size within groups (Jobling, 1994), and minimize feed wastage and water pollution (Puvanendran et al., 2003). The objective of this work was to evaluate the ideal feeding rate and frequency for juvenile pompano. Materials and Methods Juvenile pompano were captured in shallow coastal waters at Cassino beach, RS, in the South region of Brazil (32°12'S, 52°10'W), using a beach seine net (2.5x1.5 m, 5.0 mm mesh opening), and were taken to the Laboratório de Piscicultura Estuarina e Marinha, at the Universidade Federal do Rio Grande, in March 2007. Fish were treated with a prophylactic formalin bath (100 ppm) for 1 hour, in order to remove external parasites, and were acclimatized in a 1.000 L tank for one month before the beginning of the experiments. Fish were fed until visual satiation three times a day with a NRD commercial diet (INVE Aquaculture, Inc., Salt Lake City, UT, USA) (59% protein, 16% lipid, 1 mm diameter). The tank was siphoned to remove uneaten feed and feces, and at least 90% of the water was exchanged every day. The following parameters were measured daily: water temperature and dissolved oxygen, using an oxygen meter YSI 55, (YSI Incorporated, Yellow Springs, OH, USA); salinity, with a hand‑held refractometer S/Mill‑E, (Atago Co., Tokyo, Japan); pH, using a pH meter Q400A, (Quimis, Diadema, SP, Brazil); and total ammonia nitrogen (TA‑N), using the method described by the American Public Health Association (2005). With the exception of temperature, which averaged 24.2±0.2°C and 25.0±0.1°C for experiment I and II, respectively, the other parameters did not differ among treatments and experiments: dissolved oxygen (5.38±0.1 mg L‑1 O2), salinity (35±0 g L‑1), pH (7.77±0.3), and TA‑N (1.05±0 mg L‑1 TA‑N). Both experiments were carried out simultaneously in a completely randomized design. 951 In experiment I, ive feeding levels were evaluated, all with three replicates: 4, 8, 12, 16, and 20% body weight per day. The experiment lasted for 21 days, during which 25 ish (4.8±0.6 g and 6.48±0.01 cm) were stocked in 15 cylindrical tanks (50 L). To prevent escape, all tanks were covered with mesh. Fish were fed six times a day (at 2, 6, 10, 14, 18, and 22h). In experiment II, four feeding frequencies were tested during 28 days: 2 (0 and 12h), 6 (0, 4, 8, 12, 16, and 20h), 8 (0, 3, 6, 9, 12, 15, 18, and 21h), and 10 (0h, 2h30, 5h, 7h30, 10h, 12h30, 15h, 17h30, 20h, and 22h30) times a day, with three replicates each. Twenty ish (4.1±0.1 g and 6.6±0.1 cm) were randomly stocked in 15 cylindrical‑conical tanks (40 L). For all treatments, ish were fed 12% body weight per day. At the beginning of the experiment and every week thereafter, 15 ish from each tank were sampled, anesthetized with benzocaine (50 ppm) (Okamoto et al., 2009), and individually measured and weighed. Fish were returned to their respective tanks after these procedures and no mortality was observed following ish manipulation. Fish were not fed for 24 hours before measurements were taken. After this, biomass was calculated and the amount of feed offered was adjusted. At the end of the experimental period, the following zootechnical parameters were calculated: speciic growth rate, G = 100 × [(ln Wf ‑ ln Wi) / day], in which Wi and Wf are the initial and inal mean weight, respectively; apparent feed conversion, AFC = dry feed offered (g) / biomass gain (g); weight coeficient of variation, CV = (standard deviation / mean) × 100; survival, S = [(Ni ‑ Nf) / Ni] × 100, in which Ni is the initial number of ish and Nf is the inal number of ish; and condition factor, CF = BW / L3, in which BW is the body weight (mg) and L is the length (cm). The water and zootechnical parameters of both experiments were analyzed by one‑way analysis of variance followed by Tukey’s test, at 5% probability. All statistical procedures were performed with the aid of the Statistica 7.0 software (StatSoft, Inc., Tulsa, OK, USA). Results and Discussion The feeding rates selected did not affect survival (Table 1). Final weight and length were signiicantly lower for ish fed 4% body weight per day than 8, 12, 16, and 20% body weight per day, and the maximum Pesq. agropec. bras., Brasília, v.48, n.8, p.950-954, ago. 2013 DOI: 10.1590/S0100-204X2013000800020 952 V.L. da Cunha et al. mean weight was observed for ish fed 8% body weight per day. Although apparent feed conversion (AFC) showed signiicant differences between treatments, with better results when feeding rate was decreased (Figure 1), there was no effect on speciic growth rate (SGR) and condition factor, which were similar among feeding rates. Optimum feeding rate is an important factor for promoting better growth and developing an appropriate feeding program (Mihelakakis et al., 2002; Cho et al., 2007). In the present study, the optimum feeding rate for juvenile pompano was 8% body weight per day, since growth was lower in ish fed with 4% body weight per day, and feeding rate above 8% body weight per day did not improve growth. The optimal feeding Table 1. Performance of juvenile pompano (Trachinotus marginatus) fed with ive different feeding rates during 21 days(1). Feeding rate (% body weight per day) 4 8 12 16 20 Survival (%) 100±0.0 98.7±1.3 97.3±2.7 100±0.0 100±0.0 Final weight (g) 8.36±0.2b 9.68±0.2a 9.64±0.2a 9.56±0.2a 9.61±0.2a CV weight 19.8±3.4 18.6±5.0 17.3±4.1 18.1±4.4 18.0±2.8 Final length (cm) 7.87±0.1b 8.33±0.1a 8.28±0.1a 8.25±0.1a 8.29±0.1a AFC 1.54±0.1d 2.52±0.1cd 3.56±0.2c 4.85±0.2b 5.98±0.3a SGR (%) 2.63±0.2 3.13±0.2 3.31±0.2 3.37±0.1 3.28±0.2 CF 1.70±0.0 1.66±0.0 1.68±0.0 1.69±0.0 1.67±0.0 Parameter CV, coeficient of variation; AFC, apparent feed conversion; SGR, speci‑ ic growth rate; CF, condition factor. Mean±SE from three replicates from each treatment. Means followed by different letters, in the lines, differ by Tukey’s test, at 5% probability. (1) 7 Apparent feed conversion 6 5 4 3 2 y = 0.2804x + 0.3255 2 R = 0.96 P = 0.0000 n = 25 1 0 4 8 16 12 Feeding rate (% body wieght per day) 20 Figure 1. Apparent feed conversion of juvenile pompano (Trachinotus marginatus) as a function of feeding rate. Pesq. agropec. bras., Brasília, v.48, n.8, p.950‑954, ago. 2013 DOI: 10.1590/S0100‑204X2013000800020 rate found for juvenile pompano (8% body weight per day) was high when compared to those reported for others subtropical species, such as Sparus aurata (Mihelakakis et al., 2002), Paralichthys olivaceus (Kim et al., 2007), and Limanda ferruginea (Puvanendran et al., 2003), of 2.3, 2.6, and 3.0% body weight per day, respectively. The feeding rate of juvenile pompano seems to be closer to values found for tropical ish, including Clarias gariepinus (8% body weight per day) (Marimuthu et al., 2011) and Colossoma macropomum (10% body weight per day) (Silva et al., 2007), which is probably due to the high metabolism of T. marginatus (Cunha et al., 2009). The eficiency in converting feed into increased body mass is evaluated by AFC measurements. Increase in feeding rate affected AFC of pompano, and the best results were observed at 4 and 8% body weight per day, whereas ish fed 12, 16, and 20% body weight per day showed worse AFC. The best AFC at 4% body weight per day may have been a consequence of restricted feeding, when ish tend to optimize digestion in order to use nutrients more eficiently, improving feed eficiency (Mihelakakis et al., 2002; Kim et al., 2007). However, the increase of feeding rate did not improve SGR, which was statistically similar for all treatments. An opposite result was obtained by Sun et al. (2006), for juvenile cobia (Rachycentron canadum), and Silva et al. (2007), for juvenile tambaqui (C. macropomum), who found that an increased feeding rate increased SGR. In general, the maximum SGR and maximum feeding eficiency do not occur at the same feeding rate (Puvanendran et al., 2003). Furthermore, better growth following high AFC is an indicator of overfeeding, which results in higher production costs, water pollution, and feed wastage, and, therefore, should be avoided (Mihelakakis et al., 2002; Cho et al., 2007; Kim et al., 2007). According to Puvanendran et al. (2003), the establishment of hierarchy in rearing tanks is common and can cause variability in ish size. Although this experiment was not designed to evaluate behavior, it was observed that pompano fed the lowest feeding rate showed aggressive behavior; however, the coeficient of variation of body weight was similar for all treatments. Therefore, ish size, physiological state, circadian cycle, feed composition, and rearing practices also need to be taken into account because they play an Feeding rate and frequency on juvenile pompano growth important role on the determination of the optimum feeding rate (Jobling, 1994; Heilman & Spieler, 1999; Puvanendran et al., 2003; Wang et al., 2007). The tested feeding frequency did not affect survival (Table 2). Final weight was signiicantly higher for ish fed 10 times a day when compared to ish fed 6 and 2 times a day. However, ish fed 8 times a day did not show differences among all treatments. Moreover, no signiicant differences for length among feeding frequencies were detected. Pompano fed 10 and 8 times a day had higher SGR than that those fed 2 and 6 times a day. AFC was signiicantly higher for pompano fed 2 times a day than for those fed 10 and 8 times a day, but did not differ signiicantly between ish fed 2 and 6 times a day. No signiicant differences were observed regarding the condition factor among treatments. Feeding frequency has a strong inluence on ish growth performance and should provide the quantities of food to support good growth (Lee et al., 2000; Wang et al., 1998). The effect of feeding frequency on survival appears to be species‑speciic, since, as observed in the present study and for Sebastes schlegeli (Lee et al., 2000), the survival rate was not affected by feeding frequency. However, for juvenile Pagellus erythrinus, survival decreases when feeding frequency is decreased from 4 to 1 times a day (Mihelakakis et al., 2001). Studies on feeding frequency have shown that weight gain increases when the feeding frequency is increased up to a certain limit, and that a frequency increase above this limit would result in unnecessary costs (Mihelakakis et al., 2001; Wang et al., 2007). Table 2. Performance of juvenile pompano (Trachinotus marginatus) fed with four different feeding frequencies during 28 days(1). Parameter Survival (%) Final weight (g) CV weight Final length (cm) AFC SGR (%) CF 2 96.7±1.7 8.58±0.3b 23.9±3.3 7.37±0.1 4.36±0.1a 2.61±0.1b 1.86±0.1 Feeding frequency (times a day) 6 8 10 100±10.0 96.7±1.7 96.7±1.7 8.78±0.2b 9.28±0.4ab 9.79±0.3a 20.9±3.5 21.1±2.6 24.5±1.2 7.76±0.1 7.86±0.1 8.02±0.1 4.20±0.2ab 3.61±0.3b 3.50±0.1b 2.61±0.1b 3.19±0.1a 3.29±0.1a 1.87±0.1 1.89±0.1 1.90±0.1 CV, coeficient of variation; AFC, apparent feed conversion; SGR, speci‑ ic growth rate; CF, condition factor. Mean±SE from three replicates from each treatment. Means followed by different letters, in the lines, differ by Tukey’s test, at 5% probability. (1) 953 For juvenile pompano, this limit corresponds to 8 times a day, since increasing the frequency to 10 times a day did not improve growth. Wang et al. (1998) reported decreasing weight coeficient of variation for hybrid sunish (female green sunish Lepomis cyanellus x male bluegill L. macrochirus) at an increasing feeding frequency, suggesting a more uniform growth at higher feeding frequencies. However, this was not observed for juvenile pompano, which showed no signiicant difference as to the coeficient of variation in weight among treatments. Riche et al. (2004) found that feeding frequency is strongly inluenced by time of gastric evacuation, which is useful for assessing appetite return and, therefore, can be used as a tool to estimate proper feeding frequency (Lee et al., 2000). Cunha et al. (2009) suggested that the process of digestion and assimilation of food by juvenile pompano lasts for 2.5 hours, corresponding exactly to 8 times a day as observed in the present study. According to these results, it seems appropriated to feed juvenile pompano 8 times a day. Conclusions 1. 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Effect of feeding strategies on production characteristics and body composition of Florida pompano reared in marina recirculating systems. North American Journal of Aquaculture, v.68, p.330‑338, 2006. DOI: 10.1577/A05‑082.1. Received on August 1st, 2011 and accepted on July 22, 2013 Pesq. agropec. bras., Brasília, v.48, n.8, p.950‑954, ago. 2013 DOI: 10.1590/S0100‑204X2013000800020