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A feeding trial was conducted to evaluate the effects of two different sizes of extruded pellets (EP) (EP - 3 mm or EP - 5 mm) and a moist pellet (MP) in olive flounder, Paralichthys olivaceus, reared in semi-recirculation system. A total of 450 fish with an average initial weight of 5.0 ± 0.2 g (mean ± SD) were fed one of the three experimental diets in triplicate groups. At the end of a 6-week feeding trial, weight gain, specific growth rate, and feed efficiency of fish fed EP diets were significantly higher than those of fish fed MP (P < 0.05). Water quality parameters like turbidity, total ammonia nitrogen, and total phosphorous from tanks of fish fed EP and EP were significantly lower 1 2 than those from tanks of fish fed MP. Blood plasma glutamic oxaloacetic transaminase and glucose concentration were significantly higher in fish fed MP diet compared to fish fed EP diets (P < 0.05). Whole body crude protein contents in fish fed EP diets were higher than those from the fish fed MP diet. Whole body amino acid content like threonine, aspartic acid, serine, tyrosine, and cystine were found to be significantly higher in fish fed EP diets than those in fish fed MP diet. In considering overall performance of olive flounder, EP diet could be recommended for the successful aquaculture of this important fish species. Keywords: Extruded pellet, Moist pellet, Growth, Hematology, Juvenile olive flounder Background fish can ingest over a period of time. Undesirable size of Fish feeding is one of the most important factors in pellets or a high amount of pellets may cause feed wast- commercial fish farming because feeding regime may age, as fish may be unable to ingest the required amount have consequences on both growth performance and of feeds (Bailey et al. 2003). feed wastage (Tsevis et al. 1992; Azzaydi et al. 2000). Olive flounder, Paralichthys olivaceus, is one of the During the last decade, there has been a marked increase most commercially important marine aquaculture spe- in the use of extruded pellet (EP) for feeding fish. It has cies in Korea. Production of the olive flounder was been well documented that EP diets have superior water 42,133 metric tons, ranked it first among Korean mari- stability, better floating properties, and higher energy culture finfish species in 2014 (KOSTAT 2015). The suit- content than other pelleted diets (Hilton et al. 1981; able type and size of pellets for different age groups of Johnsen and Wandsvik 1991; Ammar 2008). However, olive flounder are very important for maximum growth. the size of feed pellets and the rate at which they are de- Most of the flounder production has been sourced from livered may affect the amount of feed that an individual use of frozen raw fish (sardine or mackerel) or raw fish- based moist pellets (MP) composed of frozen raw fish and commercially available binder meal at a certain ratio * Correspondence: firstname.lastname@example.org Department of Marine Bio-materials and Aquaculture/Feeds and Foods (Cho and Cho 2009). Nutrition Research Center (FFNRC), Pukyong National University, Busan There are several studies that have been done in the 608-737, Republic of Korea context of nutrient requirements and feeding technology Full list of author information is available at the end of the article © 2016 The Author(s). Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Lee et al. Fisheries and Aquatic Sciences (2016) 19:32 Page 2 of 6 of olive flounder. However, information on the effects of Water quality analysis extruded pellets and their size for juvenile olive flounder Water samples from the fish tanks were monitored just is scarce. Therefore, the present experiment was con- after 2 h of feeding. Turbidity, total ammonia nitrogen ducted to evaluate the effects of two different sizes of (TAN), and total phosphorous (TP) were determined from extruded pellets and a moist pellet on the water quality, the water of the each experimental tank. The concentration growth performance, body composition, hematological of turbidity, TAN, and TP were recorded according to the characteristics, and gut histology of juvenile olive floun- standard methods for marine environmental analysis der, P. olivaceus. (Ministry of Land Transport and Maritime Affairs 2010). Sample collection and analysis Methods At the end of the feeding trial, fish were starved for 24 h Experimental diets and they were counted and weighed to calculate the In this study, the MP diet was prepared from frozen sar- weight gain (WG), specific growth rate (SGR), feed effi- dine and commercial wheat flour at the ratio of 3:1 (wet ciency (FE), and survival rate. After the final weighing, weight basis) and the EP (3 mm) and EP (5 mm) feeds three fish from each aquarium were analyzed for whole 1 2 were provided by EWOS Canada Ltd. EP was designed body proximate composition. Proximate composition of to maintain the same level of protein (56 %) and lipid the experimental diets and fish bodies were performed (10 %). Proximate compositions of the experimental di- by the standard methods of AOAC (1995). To determine ets are shown in Table 1. Diets were stored at −20 °C the moisture content of diets and fish, they were dried (wet pellets) until use. to maintain constant weight at 135 °C for 2 h. Ash con- tent was determined using a muffle furnace (550 °C for 4 h). Crude lipid content was determined by the soxhlet Experimental fish and feeding trial extraction method by using Soxtec system 1046 (Foss, Juvenile olive flounder, P. olivaceus, was obtained from Hoganas, Sweden) and crude protein content by Kjeldahl Tong-yeong, Republic of Korea. Prior to the feeding trial, method (N × 6.25) after acid digestion, distillation, and ti- the fish were fed different experimental diets for 2 weeks to tration of samples. Fiber content was analyzed with a fiber allow them to adjust to the experimental diets and condi- analyzer (FT122 Fibertec™, Foss, Hillerød, Denmark). tions. Feeding trial was conducted in a semi re-circulating Carbohydrate content was calculated by subtracting total system with 250-L aquaria each having a water flow rate of percentage of nutrient contents from 100 %. Gross energy 1.5 L/min. Supplemental aeration was provided to maintain of experimental diets were calculated based on the calcu- dissolved oxygen near saturation. Water temperature was lation of 16.7, 16.7, and 37.7 kJ/g for protein, carbohy- maintained at 21 ± 1 °C (mean ± SD). Salinity was main- drate, and lipid, respectively (Halver and Hardy 2002). tained at 31 ± 1 g/L (mean ± SD). Fish averaging 5.0 ± 0.2 g Blood samples were taken by using heparinized syrin- (mean ± SD) randomly distributed to each aquarium as ges from the caudal vein of five randomly chosen fish groups of 50 fish and fed the experimental diets in triplicate per tank. Plasma was collected after centrifugation at at a rate of 2.5~4.5 % wet body weight per day for 6 weeks. 3000rpm for10min andstoredat −70 °C in order Total fish weight in each aquarium was determined every to analyze glutamic oxaloacetic transaminase (GOT), 2 weeks, and the amount of diets fed to fish was adjusted glutamic pyruvic transaminase (GPT), glucose, total pro- accordingly. tein (T-protein), cholesterol, and triglycerides. Plasma ana- lyses were performed at the National Fisheries Research Table 1 Proximate composition of the experimental diet and Development Institute (NFRDI), Gijang, Busan, (percentage dry matter basis) Korea, by using the kits of DRI-CHEM 4000i- Fuji Items Diets Dri-Chem Slide- 3150 (Minato-ku, Tokyo, Japan). Amino a a b acid analysis of edible body parts was performed by nin- EP EP MP 1 2 hydrin method (Sykam Amino Acid Analyzer S433, Moisture (%) 7.39 7.17 65.15 Sykam, Eresing, Germany). Crude ash (%) 10.91 11.07 12.08 Crude lipid (%) 10.18 10.04 10.40 Statistical analysis Crude protein (%) 56.79 56.91 57.07 All data were analyzed by one-way ANOVA (Statistix Crude fiber (%) 2.38 2.30 1.61 3.1; Analytical Software, St. Paul, MN, USA) to test the Carbohydrate (%) 14.35 14.34 13.47 effects of the dietary treatments. When a significant treatment effect was observed, an LSD test was used Gross energy (kcal/g) 3.76 3.75 3.76 a to compare means. Treatment effects were considered EP extruded pellet; EWOS Canada Ltd. MP moist pellet; composed of frozen sardine and wheat flour (3:1) at P < 0.05 level of significance. Lee et al. Fisheries and Aquatic Sciences (2016) 19:32 Page 3 of 6 Results and discussion Table 3 Growth performances of juvenile olive flounder fed with the different experimental diets for 6 weeks Water quality has been acknowledged to have profound effects on the growth performance and health of aqua- Variables Diets Pooled SEM culture fish species. In the present experiment, water EP EP MP 1 2 § b a b quality parameters were affected by the experimental di- WG 130.8 142.1 128.4 2.01 ets (Table 2). The observed water quality parameters es- ¶ b a b SGR 2.78 2.95 2.75 0.03 pecially turbidity was significantly lower among the ¤ b a b FE 101.1 103.1 101.4 0.41 group of fish fed extruded pellet diets. Turbidity caused SR* 97.3 96.0 94.0 1.15 by suspended solids has been reported to have great ef- Values are means from groups (n = 3) of fish where the values in each row fects on fish metabolism in terms of fish growth and sur- with different superscripts are significantly different (P < 0.05) vival. After 2 h of feeding, turbidity, TAN, and TP were Pooled SEM pooled standard error of mean (SD/√n) Weight gain (%): (final wt. − initial wt.) × 100/initial wt. recorded to be significantly higher in the group of fish Specific growth rate (%/day): (ln final wt. − ln initial wt.) × 100/days fed MP diet than those of fish fed EP diets. These results Feed efficiency (%): (wet weight gain/dry feed intake) × 100 may indicate that MP diet was easily soluble in water be- *Survival rate = (total fish − dead fish) × 100/total fish fore consumption by fish, whereas extruded pellets were more stable in water and their leaching rate in water was performance in flounder aquaculture. In our study, the comparatively prolonged. Folke and Kautsky (1989) re- results of lower feed efficiency in fish fed MP diet could ported that water pollution by fish feeding is caused be due to high leaching properties of the MP diet before largely by increasing turbidity, as well as ammonia and the ingestion of feed by the fish. It is well documented phosphorus loading through uneaten feeds and feces. that extruded pellets have superior water stability, better From an on-farm experiment with flounder, Kim and floating properties, and a higher energy content among Lee (2000) reported that the excretion of nitrogen (N) the pelleted diets (Hilton et al. 1981; Johnsen and ranged from 48 to 70 g and phosphorus (P) from 10 to Wandsvik 1991; Ammar 2008). Aqua feed technology is 12 g per kilogram weight gain. However, under practical moving in tandem with the aquaculture growth with the feeding conditions, flounder excreted much higher N usage of extrusion procedures for the improvement of (114 g) and P (28 g) per kilogram weight gain, suggesting digestibility (Umar et al. 2013). Chang and Wang (1999) a substantial waste of feed (Kim et al. 2002). Likewise, stated the advantages of extrusion cooking process for similar findings have been reported by Cha et al. (2008) aquaculture feed production including improved feed in their experiment with olive flounder fed MP and conversion ratio, control of pellet density, greater feed chitosan-based extruded pellets. stability in water, better production efficiency, and versa- In the present study, significantly higher growth per- tility. During extrusion cooking, various reactions take formance was observed for the group of fish fed EP place including thermal treatment, gelatinization, protein than those of fish fed MP diet (Table 3). At the end of denaturation, hydration, texture alteration, partial dehy- experiment, WG, SGR, and FE of fish fed EP diet were dration, and destruction of microorganisms and other significantly higher than those of fish fed EP and MP toxic compounds (Kannadhason et al. 2011). According diet. However, there were no significant differences in to Chang and Wang (1999), the gelatinization that oc- these parameters among the fish fed EP and MP diets. curs during extrusion process improves durability of the Survival rate ranged from 94 to 97 % without any statis- feed rations and digestibility of starch. In the present tical differences among different treatments. Likewise, study, result for the fish fed EP concordantly supported Cho and Cho (2009) reported that an extruded pellet is the various reports that extruded pellets are having a more recommendable than a moist pellet for the growth better efficiency over MP diet for juvenile olive flounder growth. However, it is difficult to attribute any reason for the observed lower weight gain for the group of fish Table 2 Water quality parameters after 2 h of feeding of fed EP diet in the present experiment. juvenile olive flounder fed with the different experimental diets The present experiment clearly demonstrated the for 6 weeks beneficial effects of pellet size of extruded pellets on the Parameters Diets Pooled SEM performance of olive flounder. Interestingly, we observed EP EP MP 1 2 a lower growth rate for the group of fish fed EP than b b a 1 Turbidity (NTU) 0.57 0.56 2.63 0.34 did fish fed EP diet. Feed pellet size will obviously have b b a TAN (mg/L) 0.33 0.31 0.53 0.04 an effect on fish performance, and there are indications b b a TP (mg/L) 0.04 0.04 0.07 0.01 of this effect presented in the study. Usually, for a pellet Value are means from groups (n = 3) of samples where the values in each row that is larger than the mouth gape of fish, handling time with different superscripts are significantly different (P < 0.05) becomes a limiting factor in the fish’s ability to ingest TAN total ammonia nitrogen, TP total phosphorus, Pooled SEM pooled standard error of mean (SD/√n) enough pellets to maintain good growth which will Lee et al. Fisheries and Aquatic Sciences (2016) 19:32 Page 4 of 6 clearly have negative effects. However, in our study, EP Table 4 Whole body proximate composition of juvenile olive flounder fed with the different experimental diets for 6 weeks feed was well accepted by the fish even though its size (percentage of DM basis) was larger than EP probably because EP was more 1 2 Items Diets Pooled suitable in relation to mouth gape size of fish than those SEM of EP diet. EP EP MP 1 2 a b a It is recommended that pellet size should be approxi- Moisture 76.87 75.36 77.11 0.30 mately 20–30 % of the size of the fish species mouth Crude ash 17.53 16.96 18.45 0.47 gape (Craig 2009). Feeding too small a pellet results in b ab a Crude lipid 3.25 5.34 7.50 0.58 inefficient feeding because more energy is used in find- a a b Crude protein 74.70 74.16 71.91 0.53 ing and eating more pellets. Conversely, pellets that are Value are means from groups (n = 3) of fish where the values in each row with too large will depress feeding and, in the extreme, cause different superscripts are significantly different (P < 0.05) choking. Therefore, it is better to select the largest sized Pooled SEM pooled standard error of mean (SD/√n) feed that the fish will actively eat. Smith et al. (1995) re- ported that the length and diameter of pellets influence various previous experiments. For instance, Cho and the detectability and/or attractiveness of pellets to sal- Cho (2009) reported from their experiments that prox- monids. In another report, Irwin et al. (2002) reported imate composition of whole body of flounder with and that smaller sized turbot prefer to accept a bigger size of without liver, except for moisture content of liver, was pellet (pellet size, 40 % of mouth gape) whereas the lar- not significantly affected by the different diets (extruded ger fish group prefers a smaller pellet size (pellet size, pellets, semi-moist pellets, and moist pellets). Results for 20 % of mouth gape) which is higher than the preferred whole body amino acids (Table 5) showing only four pellet size of salmonid species (Wankowski and Thorpe amino acids viz. aspartic acid (Asp), threonine (Thr), 1979; Brannas and Alanara 1992). The results may sup- serine (Ser), and tyrosine (Tyr) were significantly lowest port the findings of the present study. However, feed for the fish fed MP diet than those of fish fed other range selectivity may be governed by hunger levels of fish (Croy and Hughes 1991). Ellis et al. (1997) reported Table 5 Whole body amino acid composition of juvenile olive that farmed turbot prefer pellets and due to their ex- flounder fed with the different experimental diets for 6 weeks tended jaws they can engulf large prey items (Holmes (percentage of DM basis) and Gibson 1986). Some workers (Hjertnes et al. 1993; Amino acids Diets Pooled Tuene and Nortvedt 1995) have used larger pellet sizes (AA) SEM EP EP MP 1 2 in experiments with halibut than those recommended Essential (EAA) for Atlantic salmon, possibly because halibut have a lar- Methionine 1.64 1.50 1.37 0.05 ger mouth gape than salmonids of the same weight. In Leucine 5.07 4.84 4.82 0.05 contrast, Stradmeyer et al. (1988) reported that adult sal- mon showed a more immediate response to larger pel- Isoleucine 3.38 2.90 2.98 0.11 lets but that these were more likely to be rejected than Arginine 4.37 4.39 4.01 0.06 pellets of a shorter length. However, the texture and Histidine 1.87 1.78 1.90 0.04 hardness of pellets is an important issue. It has been Lysine 5.56 5.19 5.25 0.06 seen that juvenile salmon can handle larger size of soft a ab b Phenylalanine 2.72 2.64 2.54 0.03 pellets than hard pellets (Mearns 1990). Tuene and a a b Threonine 2.90 2.82 2.18 0.09 Nortvedt (1995) fed 9-mm pellets to 90–662-g halibut and concluded that the high intra-individual (day-to-day) coef- Valine 3.39 3.27 3.35 0.03 ficient of variation of feed intake may have been caused by Non-essential (NEAA) the large pellet size since the average consumption at each Alanine 4.59 4.71 4.33 0.07 meal was less than two pellets per fish. Glycine 5.31 6.06 5.24 0.18 Whole body proximate composition data revealed sig- a a b Aspartate 7.25 7.14 6.27 0.13 nificantly lower whole body crude protein contents for Proline 3.44 3.69 3.33 0.07 the group of fish fed MP diet than those of fish fed EP a a b diet, whereas whole body crude lipid content was signifi- Serine 2.93 2.87 1.50 0.20 cantly higher among the group of fish fed MP diet Glutamate 10.32 10.00 9.96 0.09 (Table 4). Moisture contents for fish fed EP were sig- a a b Tyrosine 1.94 1.83 1.08 0.12 nificantly lower than those of fish fed all other diets. a ab b Cystine 1.29 1.15 1.00 0.04 However, there was no significant difference in whole Value are means from groups (n = 3) of fish where the values in each row with body moisture content among the group of fish fed EP different superscripts are significantly different (P < 0.05) and MP diets. Similar findings have been reported in Pooled SEM pooled standard error of mean (SD/√n) Lee et al. Fisheries and Aquatic Sciences (2016) 19:32 Page 5 of 6 experimental diets. Although significant differences were be a right choice to minimize water pollution and in- recorded in whole body amino acids for other 13 amino crease total production in flounder aquaculture. acids, no clear trend could be drawn among different treatments. MP diet appeared to affect distinctly only Conclusions these four amino acids. Due to the lack of reports on The results from the present study demonstrated the whole body amino acid contents in similar studies, it is beneficial effects of EP and their diameter over com- difficult to compare the present observation with others. monly used MP diets in promoting growth of olive Hematological characteristics can be used as an index of flounder, suggesting the need to revise the feed and feed- the health status of fish (Blaxhall 1972). Hematological ing technology for flounder aquaculture. In the present changes have been detected following different types of experiment, the results evidenced that fish fed EP stress conditions like exposure to pollutants, diseases, and (5 mm) had the better growth and water quality parame- hypoxia (Duthie and Tort 1985). Hence, it could be sug- ters than did fish fed MP diet in juvenile olive flounder. gested that any unhealthy condition caused by poor nutri- Abbreviations tion could affect the hematological characteristics of fish. CL: Crude lipid; CP: Crude protein; EAA: Essential amino acids; EP: Extruded Plasma glucose concentration is one of the stress indica- pellet; FE: Feed efficiency; GOT: Glutamic oxaloacetic transaminase; GPT: Glutamic pyruvic transaminase; MP: Moist pellet; SGR: Specific growth tors in fish (Menezes et al. 2006) which may vary greatly rate; SR: Survival rate; WG: Weight gain depending on the physiological status of the animal (de Andrade et al. 2007). Mommsen et al. (1999) reported that Acknowledgements plasma glucose values can increase, decrease, or keep con- We would like to acknowledge the members of the Feeds and Foods Nutrition Research Center (FFNRC), Pukyong National University, Busan, Republic of Korea, stant under a high plasma cortisol level. Plasma GOT and for their assistance. GPT activities may give information on liver injury or dys- function (Wells et al. 1986). They are also used as valuable Funding This research work has been financially supported by EWOS Canada Ltd. diagnostic means of stress responses in several fish species (Almeida et al. 2002; Choi et al. 2007). The present Availability of data and materials study has been indicated that plasma GOT and glu- The datasets supporting the conclusions of this article are included within the article. There is no additional data and materials to disclose. cose in the group of fish fed MP diet were signifi- cantly higher than those of fish fed EP diet because Authors’ contributions fish might be always in stress of feed competition SL conducted the research, analyzed the samples, and prepared the draft manuscript. MM helped to write the draft manuscript. JB, MS, and YS helped (Table 6). However, no significant differences were in the research conduction and statistical analysis. BKD helped in the found in GPT, T-protein, cholesterol, and triglyceride research design and reviewed the manuscript. SCB designed and monitored levels among the fish fed EP or MP diet. the experiment and finalized the draft manuscript. All authors read and approved the final manuscript. In salmon aquaculture, MP diets were used due to their better acceptance with soft texture and relatively Competing interests low cost compared to dry diet (Ghittino 1979). However, The authors declare that they have no competing interests. in yellow tail and flounder culture, MP diet has demerits Consent for publication in causing water pollution from leftover feed which ul- The manuscript has been read and approved by the authors, and none of its timately increases production cost by decreasing water parts have been submitted and published elsewhere. The authors also quality and quantity of fish (Kim and Shin 2006; Kim declare that nobody who qualifies for authorship has been excluded from the list of authors. et al. 2007). In this instance, extruded pellet diet could Ethics approval Table 6 Hematological parameters of juvenile olive flounder The experiment was conducted under the guidelines of Animal Ethics fed with the different experimental diets for 6 weeks Committee Regulations, No. 554, issued by the Pukyong National University, Busan, Republic of Korea. Parameters Diets Pooled SEM EP EP MP 1 2 Author details b b a 1 GOT (U/L) 34.0 34.7 44.9 1.96 Department of Marine Bio-materials and Aquaculture/Feeds and Foods Nutrition Research Center (FFNRC), Pukyong National University, Busan GPT (U/L) 5.0 5.3 5.0 0.23 608-737, Republic of Korea. EWOS Canada Ltd., 7721-132nd Street Surrey, b b a Vancouver, British Columbia, Canada. Glucose (mg/dL) 62.0 61.0 86.3 4.18 T-protein (g/dL) 3.4 3.3 3.0 0.06 Received: 14 July 2016 Accepted: 7 October 2016 Cholesterol (mg/dL) 238.0 265.5 238.6 10.5 Triglycerides (mg/dL) 259.4 308.2 325.4 28.0 References Almeida JA, Diniz YS, Marques SG, Faine LA, Ribas BO, Burneiko RC, Novelli EB. 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Fisheries and Aquatic Sciences – Springer Journals
Published: Oct 19, 2016
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