Rahman et al.

Fisheries and Aquatic Sciences (2016) 19:15

DOI 10.1186/s41240-016-0018-8

RESEARCH ARTICLE Open Access

Effect of dietary carbohydrate sources on

apparent nutrient digestibility of olive
flounder (Paralichthys olivaceus) feed
Md Mostafizur Rahman1, Kyeong-Jun Lee2 and Sang-Min Lee1*

Abstract
Apparent digestibility coefficients (ADCs) of dry matter, crude protein, crude lipid, nitrogen-free extract, and energy
in selected carbohydrate sources including wheat flour (WF), α-potato starch (PS), α-corn starch (CS), Na alginate
(AL), dextrin (DEX), and carboxymethyl cellulose (CMC) were determined for olive flounder. The olive flounder
averaging 150 ± 8.0 g were held in 300-L tanks at a density of 30 fish per tank. Chromic oxide was used as the inert
marker. Feces were collected from the flounder by a fecal collector attached to a fish rearing tank. Apparent dry
matter and energy digestibilities of flounder fed WF, PS, CS, and DEX diets were significantly higher than those of
fish fed AL and CMC diets. Apparent crude protein digestibility coefficients of flounder fed PS and CS diets were
significantly higher than those of fish fed AL, DEX, and CMC diets. Apparent crude lipid and nitrogen-free extract
digestibility coefficients of flounder fed PS and DEX diets were significantly higher than those of fish fed WF, CS, AL,
and CMC diets. The present findings indicate that PS and DEX could be effectively used as dietary carbohydrate
energy compared to WF, CS, AL, and CMC for olive flounder.
Keywords: Flounder, Apparent digestibilities, Carbohydrate source

Background The use of dietary carbohydrate by fish depends on

Carbohydrates are valuable ingredient in formulated many factors including complexity, type, source, degree,
aquaculture diets (Rosas et al. 2000), and they can spare and heat treatment of the carbohydrate; fish species; and
the use of protein as an energy source (Simon 2009). As environmental condition (NRC 1993; Hemre et al. 2002).
the least expensive dietary energy source and good bind- However, its efficient utilization is linked to efficient di-
ing agent during the pelleting procedure (Arnesen and gestibility, and the ability of fish to digest carbohydrate
Krogdahl 1993; González-Félix et al. 2010), carbohydrate has been reported to be variable between species (Stone
utilization by various species of cultured fish is getting et al. 2003). Physiological variations associated with the
more and more attention to nutritionists and food man- feeding habits of the species, using the chemical features
ufacturers (Niu et al. 2012). Carbohydrate digestion and of the carbohydrate, lead to variable digestibility. This
capacity is variable among fishes where carnivorous fish variability demonstrates physiological and functional dif-
are less able to utilize them than omnivorous and herb- ferences of the gastrointestinal tract and associated or-
ivorous species (Krogdahl et al. 2005). In general, carbo- gans of fish species (Krogdahl et al. 2005; González-Félix
hydrate inclusion in carnivorous fish diets is limited to et al. 2010).
20 % (NRC 2011). The efficiency of dietary carbohydrate González-Félix et al. (2010) reported that carbohydrate
utilization in fish diets has been examined for many spe- digestibility in Florida pompano, Trachinotus carolinus,
cies (Peres and Oliva-Teles 2002; Stone 2003; Lee et al. is approximately 50 % emphasizing its restricted avail-
2003; Wang et al. 2005). ability and therefore confined energy digestibility. Niu
et al. (2012) investigated the dry matter and protein di-
* Correspondence: [email protected]
gestibility of some carbohydrate feedstuffs in juvenile
1
Department of Marine Biotechnology, Gangneung-Wonju National tiger shrimp, Penaeus monodon. Deng et al. (2005)
University, 7 Jukheon-gil, Gangneung, Gangwon-do 25457, South Korea determined apparent digestibility coefficients (ADCs)
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.
Rahman et al. Fisheries and Aquatic Sciences (2016) 19:15 Page 2 of 5

of hydrolyzed potato starch for juvenile white sturgeon. Table 1 Ingredients and chemical composition of the
But, no reports are available regarding the ADCs of carbo- experimental diets
hydrate sources for olive flounder. Information on appar- Diets
ent digestibility of ingredients in flounder diets is needed WF PS CS AL DEX CMC
to improve diet formulation and reduce feed production Ingredients (%)
costs. Therefore, this study was conducted to evaluate the Fish meal 61.0 61.0 61.0 61.0 61.0 61.0
ADC of different carbohydrate sources including wheat
Corn gluten meal 5.0 5.0 5.0 5.0 5.0 5.0
flour (WF), α-potato starch (PS), α-corn starch (CS), Na
alginate (AL), dextrin (DEX), and carboxymethyl cellulose Dehulled soybean meal 5.0 5.0 5.0 5.0 5.0 5.0
(CMC) for olive flounder. Wheat flour 20.0
α-potato starch 20.0
Methods α-corn starch 20.0
Experimental design and diet preparation Na alginate 20.0
A feeding experiment with three replicates was employed
Dextrin 20.0
to investigate the effects of dietary carbohydrate source on
nutrient digestibilities of olive flounder. Six experimental Carboxymethyl cellulose 20.0
diets were formulated to contain 20 % of each test ingredi- Squid liver oil 5.0 5.0 5.0 5.0 5.0 5.0
ent such as WF, PS, CS, AL, DEX, and CMC. Ingredients Vitamin premixa 1.2 1.2 1.2 1.2 1.2 1.2
and chemical composition of the experimental diets are Mineral premixb 1.5 1.5 1.5 1.5 1.5 1.5
presented in Table 1. Fish and squid liver oils were used as Vitamin C (50 %) 0.3 0.3 0.3 0.3 0.3 0.3
protein and lipid sources. Chromic oxide at a concentra-
Vitamin E (25 %) 0.2 0.2 0.2 0.2 0.2 0.2
tion of 0.5 % dry matter was added as an inert marker.
The diets were thoroughly mixed with 40 % distilled water, Choline salt (50 %) 0.3 0.3 0.3 0.3 0.3 0.3
pelleted by a wet pelleting machine, dried at room Cr2O3 0.5 0.5 0.5 0.5 0.5 0.5
temperature for 24 h, and stored at −25 °C until use. Nutrient content (dry matter basis)
Crude protein (%) 53.2 49.6 50.7 49.8 49.3 49.4
Fish and experimental condition Crude lipid (%) 7.9 7.5 8.2 9.4 9.2 8.4
The flounder were obtained from a private hatchery
Ash (%) 13.8 13.1 13.9 17.8 14.1 17.0
(Jeju Island, Korea) to the Marine and Environmental
Research Institute of Jeju National University (Jeju, N-free extract (%)c 25.1 29.8 27.2 23.0 27.4 25.2
South Korea). They were acclimated to the laboratory Gross energy (kcal/g diet) 4.2 4.0 4.2 3.7 4.2 4.0
a
condition for 2 weeks. Afterwards, the experimental Vitamin premix contained the following amount which were diluted in
cellulose (g/kg mix): DL-α-tocopheryl acetate, 18.8; thiamin hydrochloride, 2.7;
fishes (150 ± 8.0 g) were randomly distributed into riboflavin, 9.1; pyridoxine hydrochloride, 1.8; niacin, 36.4; Ca-D-pantothenate,
300-L cylindrical fiberglass tanks filled with 200 L of 12.7; myo-inositol, 181.8; D-biotin, 0.27; folic acid (98 %), 0.68; p-aminobenzoic
water at a density of 30 fish per tank in a flow- acid, 18.2; menadione, 1.8; retinyl acetate, 0.73; cholecalciferol, 0.003; and
cyanocobalamin, 0.003
through system prior to starting the digestibility test. b
Mineral premix contained the following ingredients (g/kg mix): MgSO4⋅7H2O,
The fish were hand-fed the experimental diets to vis- 80.0; NaH2PO4⋅2H2O, 370.0; KCl, 130.0; ferric citrate, 40.0; ZnSO4⋅7H2O, 20.0;
Ca-lactate, 356.5; CuCl, 0.2; AlCl3⋅6H2O, 0.15; KI, 0.15; Na2Se2O3, 0.01;
ual satiety once a day for 10 days. Filtered seawater MnSO4⋅H2O, 2.0; and CoCl2⋅6H2O, 1.0
was supplied at a flow rate of 3 L/min to each rear- c
Calculated = 100 − (crude protein + crude lipid + ash)
ing tank. Fish rearing tanks had a sloping bottom
leading to a centrally located drainage slot, and the collected from the fecal collection columns at 10:00 h
effluent water was directed first over a fecal collection for 10 days. The feces were immediately filtered with fil-
column and then to waste. Photoperiod was main- ter paper (Whatman #1) for 60 min at 4 °C and stored
tained on natural condition during the experimental at −75 °C for chemical analyses. Fecal samples from each
period. The water temperature was maintained at tank were pooled at the end of the experiment.
20.0 ± 0.82 °C.
Analytical methods
Fecal collection technique Freeze-dried feed and feces were finely ground using a
Three replicate groups of fish were carefully hand-fed grinder. Fish scales were removed using a 300-μm sieve
the test diets to visual satiety once a day at 10:00 h by before analysis. Crude protein was determined by the
the same person during the experimental period. Two Kjeldahl method using an auto Kjeldahl System (Buchi,
hours after feeding, the rearing tanks and collection col- Flawil, Switzerland). Crude lipid was analyzed with ether
umns were cleaned by sponges, and uneaten feed and extraction in a soxhlet extractor (SER 148, VELP Scienti-
fecal residues were removed. The next day, feces were fica, Milan, Italy). Moisture content was determined with
Rahman et al. Fisheries and Aquatic Sciences (2016) 19:15 Page 3 of 5

a dry oven at 105 °C for 6 h, and the ash content was de- Discussion
termined after combustion at 550 °C for 4 h in a muffle Estimation of ADC values for feedstuffs is an important
furnace. Nitrogen-free extract (NFE) was calculated by aspect in screening the nutritive value of feed ingredi-
difference. Gross energy was analyzed by an adiabatic ents and formulation of nutritionally sufficient diets
bomb calorimeter (Parr, USA). Chromic oxide content (Irvin and Tabrett 2005). In this study, dry matter digest-
of the experimental diets and fecal samples were deter- ibilities (65–78 %) of flounder fed the diets containing
mined by a wet-acid digestion method (Furukawa and different carbohydrate sources were higher compared to
Tsukahara 1966). All chemical analyses from each tank those (48–63 %) of rockfish fed the diets containing
were performed in triplicates. α-potato starch, β-potato starch, β-corn starch, and
Apparent digestibility coefficients (ADCs) for dry mat- dextrin (Lee and Pham 2011) but similar to results
ter, nutrient, and energy of the experimental diets were (68–79 %) for spiny lobster, Jasus edwardsii, fed WF,
determined using the following equations: DEX, PS, and CMC (Simon 2009). The low dry mat-
ter digestibility of CMC in this study can be attrib-
uted to the high content of ash. Lee (2002) reported
ADC of dry matter ð%Þ ¼ ð100−ðdietary Cr2 O3 =feces Cr2 O3 Þ  100Þ;
that dry matter digestibility of rockfish fed the different in-
gredients appeared to relate to the quantity and chemical
ADC of nutrients or energy composition of the carbohydrate used. It has been found
  that complexity of the carbohydrate significantly influ-
dietary Cr2 O3 feces nutrient or energy
¼ 1−   100: ences its utilization in the fish diet (Jobling 2001; Lee and
feces Cr2 O3 dietary nutrient or energy
Pham 2011). Stone et al. (2003) reported that simple
carbohydrate has high dry matter digestibility compared
to the complex kind.
Statistical analysis ADC of protein in the present study ranged from 72
All data were subjected to one-way analysis of variance to 90 %. Niu et al. (2012) reported that apparent protein
(ANOVA) using SPSS version 20.0 (SPSS Inc., Chicago, digestibility of wheat starch, sucrose, potato starch,
IL, USA). Significant differences (p < 0.05) among the maize starch, and dextrin diets ranged from 81 to 92 %
means were determined using Duncan’s multiple range for juvenile tiger shrimp, P. monodon. Apparent crude
test (Duncan 1955). Correlation of nutrients and energy protein digestibility of α-potato starch, β-potato starch,
was assessed using Pearson regression. All data were β-corn starch, and dextrin diets ranged from 90 to 95 %
presented as mean ± SE of three replicate groups. for juvenile and grower rockfish (Lee and Pham 2011).
Niu et al. (2012) demonstrated that protein digestibility
Results of potato starch diet was significantly higher than that of
Apparent nutrient digestibility of flounder fed different maize starch and dextrin diets. ADC of protein appeared
dietary carbohydrate sources is presented in Table 2. Ap- to have a positive relationship with dry matter (DM) di-
parent dry matter and energy digestibilities of flounder gestibility (r = 0.90, p < 0.01) of the test diets. In the
fed WF, PS, CS, and DEX diets were significantly higher present study, low crude protein digestibility of CMC
than those of fish fed AL and CMC diets. Apparent diet may be due to the accelerated passage of the digesta
crude protein digestibility coefficients of flounder fed PS from the stomach of olive flounder (Yamamoto and
and CS diets were significantly higher than those of fish Akiyama 1995).
fed AL, DEX, and CMC diets. Apparent crude lipid and NFE digestibility exhibited a significant correlation to
NFE digestibility coefficients of flounder fed PS and DM digestibility (r = 0.76, p < 0.01) of the test diets. Al-
DEX diets were significantly higher than those of fish though the actual mechanism responsible for poor carbo-
fed WF, CS, AL, and CMC diets. hydrate utilization in fishes has not been recognized,

Table 2 Apparent nutrient digestibility (%) of olive flounder fed different dietary carbohydrate sources
Diets Dry matter Crude protein Crude lipid NFE Gross energy
c cd c b
WF 74.5 ± 0.41 89.3 ± 0.17 81.4 ± 0.29 62.0 ± 0.61 78.8 ± 0.35c
c d e c
PS 75.4 ± 0.29 90.0 ± 0.12 89.6 ± 0.12 76.7 ± 0.29 79.7 ± 0.40c
CS 74.7 ± 0.80c 89.9 ± 0.31d 83.8 ± 0.51d 59.4 ± 1.31b 79.8 ± 0.63c
b b a b
AL 69.3 ± 0.59 78.5 ± 0.42 64.1 ± 0.71 62.4 ± 0.74 67.9 ± 0.62b
DEX 78.5 ± 3.17c 87.3 ± 1.86c 94.3 ± 0.84f 86.3 ± 2.04d 87.3 ± 1.86d
a a b a
CMC 64.6 ± 0.30 72.4 ± 0.27 71.6 ± 0.27 53.2 ± 0.43 62.0 ± 0.33a
Values (mean ± SE of three replications) in the same column not having a common superscript are significantly different (p < 0.05)
Rahman et al. Fisheries and Aquatic Sciences (2016) 19:15 Page 4 of 5

inferior digestion of carbohydrate in carnivorous fish Acknowledgements

might be attributed to the environment in which they This work was supported by a grant from the National Institute of Fisheries
Science (R2016016) in Korea.
evolved (McGoogan and Reigh 1996). Lee and Pham
(2011) reported that NFE digestibility of α-potato starch,
Authors’ contributions
β-potato starch, β-corn starch, and dextrin ranged from MMR analyzed the chemical composition and prepared the draft paper. KJL
20 to 56 % for juvenile and grower rockfish. In this study, manufactured the feed and conducted the feeding trial. SML designed this
study, the feeding system, and the revised paper. All authors read and
the apparent NFE digestibility values were 53 to 86 %. The
approved the final manuscript.
highest NFE digestibility value obtained for DEX was
86 %, and the lowest value observed for CMC was 53 %. Competing interests
This result indicates that differences in NFE digestibility The authors declare that they have no competing interests.
of the diets containing different carbohydrate sources
Author details
might be due to their indigestible polysaccharide content 1
Department of Marine Biotechnology, Gangneung-Wonju National
(Lee and Pham 2011). University, 7 Jukheon-gil, Gangneung, Gangwon-do 25457, South Korea.
2
Regarding carbohydrate sources, low NFE digestibility Department of Marine Life Sciences, Jeju National University, Jeju 63243,
South Korea.
of carbohydrates indicates that digestibility of carbohy-
drates may be attributed to their reduced solubility and Received: 12 May 2016 Accepted: 19 May 2016
to their impediment of α-amylase activity. Generally, the
fish has low ability in digestion of carbohydrates due to
its specific carbohydrase enzyme (Jobling 2001). The ef- References
Arnesen P, Krogdahl Å. Crude and pre-extruded products of wheat as nutrients
fects of solubility and α-amylase on digestibility of differ- sources in extruded diets for Atlantic salmon (Salmo salar L.) grown in sea
ent carbohydrates have been reported in silver perch water. Aquaculture. 1993;118:105–17.
(Stone et al. 2003). Some researchers suggested that car- Deng DF, Hemre GI, Storebakken T, Shiau SY, Hung SSO. Utilization of diets with
hydrolyzed potato starch, or glucose by juvenile white sturgeon (Acipenser
bohydrates are mainly digested in the interior segment transmontanus), as affected by Maillard reaction during feed processing.
of the digestive tract of fish and depends on their solu- Aquaculture. 2005;248:103–9.
bility in the fluid of the digestive tract (Fange and Grove Duncan DB. Multiple-range and multiple F-tests. Biometrics. 1955;11:1–42.
Fange R, Grove D. Digestion. In: Hoar WS, Randall DJ, Bretts JR, editors. Fish
1979; Lovell 1989). There is no available information on physiology. New York: Academic Press; 1979. p. 161–260.
NFE digestibility of olive flounder to date. Hence, studies Furukawa A, Tsukahara H. On the acid digestion method for the determination of
on the topic are necessary to elaborate the carbohydrate chromic oxide as an index substance in the study of digestibility of fish feed.
Bull Jpn Soc Sci Fish. 1966;32:502–6.
utilization of flounder. González-Félix ML, Davis DA, Rossi Jr W, Perez-Velazquez M. Evaluation of
ADC of energy ranged from 62 to 80 % in this apparent digestibility coefficient of energy of various vegetable feed
study. It has been noted that energy ADC of the diet- ingredients in Florida pompano, Trachinotus carolinus. Aquaculture.
2010;310:240–3.
ary carbohydrates such as dextrin, gelatinized wheat Hemre GI, Mommsen TP, Krogdahl A. Carbohydrates in fish nutrition: effects on
starch, glucose, raw wheat starch, and raw pea starch growth, glucose metabolism and hepatic enzymes. Aquac Nutr.
seemed to be significantly influenced by carbohydrate 2002;8:175–94.
Irvin SJ, Tabrett SJ. A novel method of collecting fecal samples from spiny
kind (Stone et al. 2003). Dextrin has been reported to lobsters. Aquaculture. 2005;243:269–72.
be a very good carbohydrate energy source and suc- Jobling M. Feed composition and analysis. In: Houlihan D, Boujard T, Jobling M,
cessfully digested by fish (Lee et al. 2003; Stone et al. editors. Food intake in fish. France: Blackwell Science; 2001.
Krogdahl A, Hamre GI, Mommsen TP. Carbohydrates in fish nutrition: digestion
2003). Lee and Pham (2011) reported that α-potato and absorption in postlarval stages. Aquac Nutr. 2005;11:103–22.
starch and dextrin appeared to be effectively digested Lee SM. Apparent digestibility coefficients of various feed ingredients for juvenile
by rockfish as carbohydrate energy source. ADC of and grower rockfish (Sebastes schlegeli). Aquaculture. 2002;207:79–95.
Lee SM, Pham MA. Effects of carbohydrate and water temperature on nutrient
energy appeared to have a positive correlation with and energy digestibility of juvenile and grower rockfish, Sebastes schlegeli.
DM digestibility (r = 0.96, p < 0.01) of the test diets. In Asian-Australas J Anim Sci. 2011;24:1615–22.
this study, ADC of energy for DEX was the highest Lee SM, Kim KD, Lall SP. Utilization of glucose maltose, dextrin and cellulose by
juvenile flounder (Paralichthys olivaceus). Aquaculture. 2003;221:427–38.
among the ingredients tested. In contrast, ADC of en- Lovell T. Digestion and metabolism. In: Lovell T, editor. Nutrition and feeding of
ergy was the lowest for CMC among those tested. fish. New York: Van Nostrand Reinhold; 1989. p. 185–203.
The poor energy digestibility of CMC in this study McGoogan BB, Reigh RC. Apparent digestibility of selected ingredients in red
drum (Sciaenops ocellatus) diets. Aquaculture. 1996;141:233–44.
may be because of insufficient non-protein energy in National Research Council. Nutrient requirements of fish. Washington: National
feeds. Research Council of the National Academy Press; 1993. p. 105.
National Research Council. Nutrient requirements of fish and shrimp.
Washington: National Research Council of the National Academics;
2011. p. 363.
Conclusion Niu J, Lin HZ, Jiang SG, Chen X, Wu KC, Tian LX, et al. Effects of seven
Among the carbohydrates tested, PS and DEX could be carbohydrate sources on juvenile Penaeus monodon growth performance,
nutrient utilization efficiency and hepatopancreas enzyme activities of
effectively used as dietary carbohydrate energy compared 6-phosphogluconate dehydrogenase, hexokinase and amylase. Anim
to WF, CS, AL, and CMC for olive flounder. Feed Sci Technol. 2012;174:86–95.
Rahman et al. Fisheries and Aquatic Sciences (2016) 19:15 Page 5 of 5

Peres H, Oliva-Teles A. Utilization of raw and gelatinized starch by European sea

bass (Dicentrarchus labrax) juveniles. Aquaculture. 2002;205:287–99.
Rosas C, Cuzon G, Gaxiola G, Arena L, Lemaire P, Soyez C, et al. Influence of
dietary carbohydrate on the metabolism of juvenile Litopenaeus stylirostris.
J Exp Mar Biol Ecol. 2000;249:181–98.
Simon CJ. The effect of carbohydrate source, inclusion level of gelatinized starch,
feed binder and fishmeal particle size on the apparent digestibility of
formulated diets for spiny lobster juveniles, Jasus edwardsii. Aquaculture.
2009;296:329–36.
Stone DAJ. Dietary carbohydrate utilization by fish. Rev Fish Sci. 2003;11:337–69.
Stone DAJ, Allan GL, Anderson AJ. Carbohydrate utilization by juvenile silver
perch, Bidyanus bidyanus (Mitchell). II. Digestibility and utilization of starch
and its breakdown products. Aquac Res. 2003;34:109–21.
Wang Y, Liu YJ, Tian LX, Du ZY, Wang JT, Wang S, et al. Effects of dietary
carbohydrate level on growth and body composition of juvenile tilapia,
Oreochromis niloticusxO.aureus. Aquac Res. 2005;36:1408–13.
Yamamoto T, Akiyama T. Effect of carboxymethyl cellulose, α-starch and wheat
gluten incorporated in diets as binders on growth, feed efficiency, and
digestive energy activity of fingerling Japanese flounder. Fish Sci.
1995;61:309–13.

Submit your next manuscript to BioMed Central

and we will help you at every step:
• We accept pre-submission inquiries
• Our selector tool helps you to find the most relevant journal
• We provide round the clock customer support
• Convenient online submission
• Thorough peer review
• Inclusion in PubMed and all major indexing services
• Maximum visibility for your research

Submit your manuscript at

www.biomedcentral.com/submit