Evaluation of gross protein and lipid requirements in formulated feed for honey gourami, Trichogaster chuna (Hamilton 1822)

Article (PDF Available)inJournal of the World Aquaculture Society · August 2018with 126 Reads
DOI: 10.1111/jwas.12557
Cite this publication
Abstract
Gross protein and fat requirements in formulated feeds designed for the honey gourami, Trichogaster chuna (initial weight 1.34–1.38 g), were investigated. Feeds containing 30%, 40%, and 50% protein and 6% and 9% lipid at each protein level were tested. Protein and energy sources used were from fishmeal, shrimp meal, clam meal, soy flour, and wheat flour. An equal mixture of crude sardine oil and groundnut oil was used as the source of lipids. Comparison of the whole gourami amino acid profiles before and after the dietary treatments indicated a relative decline in all amino acids except methionine and lysine. Fatty acid profiles of whole individuals after dietary treatment showed a substantial increase in monounsaturated fatty acids relative to the initial fatty acid profiles. No significant differences were observed in fish growth between dietary treatments (p > .05). Feeds containing 30% protein and 6% lipid were found to be adequate for normal growth, while feeds with 40% protein and 6% lipid were seen to help accelerate growth and reproduction. In this study, protein and lipid levels required for regular maintenance, sexual maturation, and spawning of aquarium‐reared gourami were established.
APPLIED STUDIES
Evaluation of gross protein and lipid requirements
in formulated feed for honey gourami, Trichogaster
chuna (Hamilton 1822)
Sukumaran Mithun
1*
| Pananghat Vijayagopal
2
| Kajal Chakraborty
2
|
Diana P. S. Chan
1
1
School of Applied Science (ASC), Temasek
Polytechnic (TP), Singapore, Singapore
2
Fish Nutrition Group, Marine Biotechnology
Division (MBTD), Central Marine Fisheries
Research Institute (CMFRI), Cochin, India
Correspondence
Vijayagopal Pananghat, Fish Nutrition Group,
Marine Biotechnology Division (MBTD),
Central Marine Fisheries Research Institute
(CMFRI), Ernakulam North P.O., Ernakulam,
Cochin 682018, Kerala, India.
Email: vgcochin@hotmail.com
*
Present address
Department of Aquatic Biology and Fisheries,
University of Kerala, Kariavattom,
Thiruvananthapuram 695581, Kerala, India.
Email: mithunsugun@gmail.com
Funding information
Department of Biotechnology , Ministry of
Science and Technology, Grant/Award
Number: DBT-CREST Award (2011-12) to VP
through BT/IN/DBT-; Department of
Biotechnology (DBT), Government of India
Gross protein and fat requirements in formulated feeds designed for
the honey gourami, Trichogaster chuna (initial weight 1.341.38 g),
were investigated. Feeds containing 30%, 40%, and 50% protein and
6% and 9% lipid at each protein level were tested. Protein and energy
sources used were from fishmeal, shrimp meal, clam meal, soy flour,
and wheat flour. An equal mixture of crude sardine oil and groundnut
oil was used as the source of lipids. Comparison of the whole gourami
amino acid profiles before and after the dietary treatments indicated
a relative decline in all amino acids except methionine and lysine.
Fatty acid profiles of whole individuals after dietary treatment
showed a substantial increase in monounsaturated fatty acids relative
to the initial fatty acid profiles. No significant differences were
observed in fish growth between dietary treatments (p>.05).Feeds
containing 30% protein and 6% lipid were found to be adequate for
normal growth, while feeds with 40% protein and 6% lipid were seen
to help accelerate growth and reproduction. In this study, protein and
lipid levels required for regular maintenance, sexual maturation, and
spawning of aquarium-reared gourami were established.
KEYWORDS
aquariculture, honey gourami, nutrition
1|INTRODUCTION
There is a dearth of information about the nutritional requirements of many widely traded ornamental fish. No stan-
dards have been established for the production and supply of feeds for this type of fish. Published data are only avail-
able for 10 freshwater ornamental species (Santamaria & Santamaria, 2011). According to Ling and Ling (2006), the
10 freshwater ornamental fish in high market demand are tetra, guppy, catfish, goldfish, platy, molly, gourami, cichlid,
Received: 23 December 2016 Revised: 26 June 2018 Accepted: 5 July 2018
DOI: 10.1111/jwas.12557
© Copyright by the World Aquaculture Society 2018
J World Aquacult Soc. 2018;115. wileyonlinelibrary.com/journal/jwas 1
loach, and arowana. From the aforementioned list, gouramis were selected as a model for nutrition studies to deter-
mine their nutritional requirements in aquariculture.
Previous reports on the gross nutritional requirements of gouramis were limited to the blue gourami, Trichogaster
trichopterus (now known as Trichopodus trichopterus), and the dwarf gourami, Trichogaster lalius (Mohanta & Subrama-
nian, 2007; Shim, Landesman, & Lam, 1989; Zuanon et al., 2013). Shim et al. (1989) showed that feed protein levels
of 35% increased growth rates, adult body mass, and the percentage of vitellogenic oocytes in T. lalius. Degani and
Gur (1992) showed that the optimal dietary protein level for another wild tropical gourami, T. leerii, ranged between
26% and 36%. Mohanta and Subramanian (2007) reported that a feed composition of 35% protein, 8% lipid, and
16.7 MJ/kg digestible energy (DE) was optimal for Trichogaster trichopterus. Zuanon et al. (2013) indicated that 37%
dietary protein was necessary for Trichogaster lalius. Berenjestanaki, Fereidouni, Ouraji, and Khalili (2014) evaluated
the effect of different lipid sources on growth and reproductive performance of the three-spot gourami,
T. trichopterus. They found that fish diets, which contained 12% lipid, in which 50% of the lipid source could be
substituted with vegetable oils, particularly canola and linseed, showed improved reproductive performance com-
pared to feeds containing only fish oil as the lipid source.
Originally, males and females of the genus Trichopodus were identified as two distinct species. This fish is now
classified under the genus Trichogaster (Vishwanath, 2010). Trichogaster chuna is variously known as Colisa chuna
(Hamilton 1822), Trichopodus sota (Hamilton 1822), and Colisa sota (Hamilton 1822). It is indigenous to the Gangetic
provinces of India, and its wild isolates are nearly absent from the aquarium sector because the entire trade popula-
tion is commercially produced. Cole, Tamaru, Bailey, and Brown (1999) summarized the biology, reproduction, and
commercial production of gouramis in general and T. trichopterus in particular.
Trichogaster chuna, known as honey dwarf gourami, red flame gourami, and dwarf fire gourami, is the smallest of
the Trichogaster species. It needs to surface the water in order to breathe because of its accessory breathing organ
called labyrinth. Females are slightly larger in size than the males and reach up to 5 cm in length. The average length
of the males is 4 cm.
In the temperate regions of northern India, T. chuna require less than 12 months to reach maturity stage. How-
ever, it could breed all year round in Singapore, and the newly hatched fry could attain sexual maturity within
4 months (Shim, Landesman, & Lam, 1987). The average lifespan of T. chuna is 4 years with proper care. They are
omnivorous and typically feed on algae, zooplankton, insect larvae, and aquatic vegetation (Mookerjee & Majumder,
1960). They are sexually dimorphic, and male coloration intensifies during breeding. Females release their eggs, which
are fertilized by the males. The males collect them in their mouths and place them in bubble nests near the surface of
the water. The males chase away the females after spawning and tend to the nests by squirting water droplets above
them. In this way, the eggs are forced downwards so that the males can pick and rearrange the eggs in the nest. The
eggs hatch within 2436 hr, and the free-swimming fry leave the nest within 3 days.
The present study evaluated the gross dietary protein and lipid content levels required for growth and breeding
of the honey gourami, Trichogaster chuna. Variations were observed in the whole-body amino acid and fatty acid pro-
files before and after experimental feeding.
2|MATERIALS AND METHODS
2.1 |Feeds and dietary treatments
Six dietary treatments containing three protein levels of 30%, 40%, and 50% and two lipid levels of 6% and 9% were
formulated according to Vijayagopal (2004). The experimental feeds were labeled as 306, 309, 406, 409, 506, and
509, whereby the first two digits in each label indicated the percentage of protein and the third digit refers to the
percentage of lipids. The ingredients used in the formulation and their approximate compositions are presented in
Table 1. The marine protein mixture was made by mixing equal quantities of fish, shrimp, and clam meals. The actual
2MITHUN ET AL.
amount of marine protein mixture in the formulation varied. Table 2 shows the ingredients and their composition in
the experimental feeds.
Feeds were prepared by blending 1 kg of dry ingredients in 180 mL tap water with a laboratory twin-screw
extruder (Basic Technologies Private Limited, Kolkata, India). The feeds were extruded as 2-mm pellets and were
crushed and sieved through sieves of the American Society for Testing and Materials to produce 1-mm crumbles. A
commercially available feed with a particle size of 1 mm was used (Otohime C2, M/s Marubeni Nisshin Feed Co. Ltd.,
Tokyo, Japan) as the control to compare the relative efficacy of the experimental feeds with those formulated using
graded levels of nutrients. The commercial feed contained krill meal, fishmeal, squid meal, potato starch, wheat flour,
brewer's yeast, licorice plant, apple extract, wheat germ, guar gum, and soy lecithin. The protein and lipid levels were
51% and 11%, respectively.
2.2 |Fish and feeding
T. chuna individuals of uniform size (1.341.38 g) were sourced from Qian Hu Fish Farm Trading, 71 Jalan Lekar,
Singapore. They were quarantined for 3 weeks prior to the start of the experiment at the Aquaculture Work Station
at Temasek Polytechnic, Singapore. During the quarantine period, commercially available freshwater ornamental fish
feed (Otohime C2) of 1-mm particle size was fed to the fish once daily. Individuals were examined for abnormal
behavior and parasite infection. Mortalities were recorded daily. Fish were manually sorted and weighed in groups of
30 using an electronic balance. They were then transferred to an experimental aquarium system equipped with a
mechanical water-recirculation filter, a biological filter, and a protein skimmer.
TABLE 1 Proximate chemical composition (% dry matter) of feed ingredients used in the experimental feeds and
their cost
Ingredients CP EE CF NFE Ash AIA Cost (INR)/kg SGD/kg
Fishmeal
1
69.11 12.09 0.26 0.32 18.22 1.21 78.74 1.57
Shrimp meal
2
68.45 4.56 2.98 6.65 17.36 1.31 82.46 1.65
Clam meal
3
67.6 7.52 9.12 0.3 15.46 1.11 137.64 2.75
Soy flour
4
59.05 0.92 5.43 27.26 7.34 0.26 89.28 1.79
Wheat flour
5
11.15 1.29 0.59 85.13 1.84 0.00 27.28 0.55
Fish oil + vegetable oil
6
99.00 27.28 0.55
Vitamin C
7
1,308.82 26.18
Mineral mixture
8
114.08 2.28
Vitamin mixture
9
165.54 3.31
Mixed carotenoids
10
11,022.36 220.45
Spirulina
11
1,377.64 27.55
Antioxidant
12
396.80 7.94
Antifungal
13
12,300.80 246.02
Lecithin
14
3,100 62.00
Note. AIA: acid-insoluble ash; CF: crude fiber; CP: crude protein; EE: ether extract or crude fat; INR: Indian rupee; NFE:
nitrogen-free extract; SGD: Singapore dollar.
1
Sardine, Sardinella longiceps, from Raj Fish Meal and Oil Co. Malpe, Mangalore, India.
2
Jawala shrimp, Acetes indicus, from
Hajee K. A. Abdul Kahader Sahib, Chennai, India.
3
Black clam, Villorita cyprinoides, meal procured from the local seafood mar-
ket, Cochin, India.
4
Sakthi Soyas, Coimbatore, India.
5
Food-grade wheat flour from the local market, Cochin, India.
6
Equimix-
ture of crude sardine oil from Kiriyathan Trading Company, Cochin, India, and groundnut oil from the local market, Cochin
India.
7
Stay-C from DSM Nutritional Technologies, India.
8
Agrimin powder from Vibrac Healthcare India, Pvt. Ltd., Mumbai,
India.
9
Supplevite-M from Sarabhai Zydus Animal Health Pvt. Ltd, Vadodara, India.
10
Mixed carotenoids 7.5% powder of
Dunaliella salina from Parry Nutraceuticals, Chennai, India.
11
Certified organic spirulina from Parry Nutraceuticals, Chennai,
India.
12
BHT from Sisco Research Laboratories, Mumbai, India.
13
Sodium metabisulphate from Nice Chemicals, Cochin, India.
14
Soy lecithin from Hi-Media, Mumbai, India.
MITHUN ET AL.3
Glass aquaria tanks, 90 cm long ×55 cm wide ×40 cm high in dimensions (V = ~200 L), were each used to hold
30 ornamental fish. Seven treatments comprising six experimental feeds and one commercial feed were each stocked
in triplicate in the study.
Feeds were provided at 4% of the body weight of the fish twice daily at 08:30 and 17:30 for 9 weeks. Uneaten
food (orts) and waste were regularly siphoned off from each tank. Mortalities were recorded daily to determine sur-
vival rates, and feed weights were adjusted accordingly. The tanks were replenished with dechlorinated fresh water
as and when needed. Water temperature, pH, and dissolved oxygen (DO) were monitored weekly using a
Y.S.I. Professional Plus multiparameter probe (Xylem Analytics, Gurgaon Haryana, India). Total ammonia nitrogen
(TAN) and nitrite were determined weekly using Hach DR 3900 spectrophotometer test kits (Hach, Loveland, CO).
The temperature range was maintained at 27.827.7C, with a pH of 6.897.56 and DO level of 7.06.5 mg L
-1
,
as well as ensuring that the TAN was between 0.044 and 0.073 mg/L. All water parameters were within the optimal
water quality range as reported for Trichogaster spp.
Fish were weighed on Days 21, 42, and 63 of the experimental period. When some fish showed signs of abdomi-
nal bulging, all individuals were left undisturbed so as to allow them to build bubble nests for breeding, after which
the impact of feed on maturation and breeding was assessed.
2.3 |Chemical analyses
The proximate composition of the experimental feeds and fish were analyzed using methods defined by Association
of Official Analytical Chemists (AOAC) (1995) whereas the dry matter (DM) was determined after drying the samples
at 105C for 24 hr in a forced air-circulating oven. Crude protein (CP), ether extract (EE), and crude fiber (CF) were
determined using Kjeltec 2300, Soxtec 2043, and Fibertec 2043 (all from Foss Analytical A/S, Hillerød, Denmark),
respectively. The nitrogen-free extract (NFE) of the diets, which represented the total amount of carbohydrates, was
calculated by subtracting CP, EE, CF, and ash from DM.
Amino acid profiles of the experimental feeds were determined as described by Heinrikson and Meredith (1984).
Briefly, 0.1 g powdered diet samples were digested with 10 mL of 6 N HCl at 110C in sealed tubes for 24 hr. Tripli-
cate samples were injected into a high-performance liquid chromatography (HPLC) system (Waters, Milford, MA)
comprising dual λabsorbance detector (Model 2487), binary pumps (Model 1525), and a Pico-Tag column. Analysis
of the data was completed using Waters Breeze GPC software (Waters, Milford, MA).
The proportion of lipids in the feed samples was obtained by cold extraction using a 2:1 chloroformmethanol
mixture (Folch, Lees, & Sloane-Stanley, 1957). Following the lipid phase isolation, the solvent was evaporated, and
TABLE 2 Ingredients in the experimental feeds
Ingredients g/kg 306 309 406 409 506 509
Marine protein mix 100 100 270 270 450 450
Soy flour 300 300 300 300 300 300
Wheat flour 510 480 350 320 180 150
Fish oil + vegetable oil 40 70 30 60 20 50
Vitamin C 555555
Mineral mixture 30 30 30 30 30 30
Vitamin mixture 333333
Mixed carotenoids 444444
Spirulina 444444
Antioxidant 111111
Antifungal 111111
Lecithin 222222
4MITHUN ET AL.
2 mL of the sample was refluxed with 5 mL of 0.5% alcoholic potassium hydroxide solution for 30 min. This was then
refluxed with 6 mL BF
3
-MeOH (Sigma-Aldrich Corp., Bangalore, India) for an additional 5 min. Refluxing was carried
out under nitrogen. The dry fatty acid methyl esters in the flask were quantitatively extracted with petroleum ether
(10 mL ×3). The extract was further washed thrice with 25 mL of double-distilled water and then filtered over anhy-
drous sodium sulfate (10 g) to remove any moisture. The solvent was then evaporated under a stream of nitrogen
gas, and 2 ul of each prepared fatty acid methyl ester was injected into a Perkin ElmerAuto-System XL gas chromato-
graph (Perkin Elmer, Waltham, MA) according to methods described by Morrison and Smith (1964).
Initial and final whole-body amino acid and fatty acid compositions were analyzed at the Agri-Food & Veterinary
Authority Laboratory in Singapore (www.ava.gov.sg). Amino acid analysis was performed with HPLC in accordance
with Technical Note 5,990-4547EN from Agilent Technologies (Santa Clara, CA). Fatty acid analysis was performed
using a gas chromatograph (Agilent Technologies, Santa Clara, CA) fitted with a flame ionization detector and in
accordance with AOAC (2005).
2.4 |Statistical analysis
One-way ANOVA was used to test for differences between the dietary treatments. Means were evaluated for signif-
icance by StudentNewmanKeuls' test (p< .05). All statistical tests were performed with SPSS v. 13.0 (IBM Corp.,
Armonk, NY).
3|RESULTS
3.1 |Proximate composition and energy of feeds
Table 3 shows the proximate analysis of protein, lipid, and calculated gross energy (GE) content of the experimental
and control feeds. GE levels ranged between 18 and 19 MJ/kg in all feeds. The protein:energy ratio varied from 1.67
to 2.74 g/MJ.
3.2 |Amino acid and fatty acid content of feeds
The most abundant amino acids, as indicated in Table 4, shown in descending order in the feeds were lysine, gluta-
mate, glycine, and alanine. The levels of histidine, arginine, and threonine were observed to be similar in all experi-
mental feeds (p> .05). The levels of all other amino acids in the feeds varied significantly (p< .05), but there were no
abnormal fluctuations.
The fatty acid profiles of the feeds (Table 5) indicated that the content of palmitic acid (16:0) was the highest of
all the saturated fatty acids. The total saturated fatty acid content increased with protein and lipid levels. Of the
monounsaturated fatty acids, oleic acid (18:1n9) was found to be the most abundant whereas linoleic acid (18:2n6)
was found to be the most abundant polyunsaturated fatty acid. EPA (20:5n3) levels were higher than those of DHA
(22:6n3) in all feeds except in feed 509, where both DHA and EPA were < 1.
3.3 |Whole-body amino acid and fatty acid content
The amino acid content was found to be lower in the fish bodies after the experiment was completed compared to
before the experiment, except for cysteine and lysine, which increased in response to all dietary treatments (Table 6).
Whole-body unsaturated fatty acid levels increased by one unit compared to the initial levels. Whole-body
monounsaturated fatty acid levels increased by approximately two units, while whole-body polyunsaturated fatty
acids showed the smallest increase relative to the initial levels (Table 7).
MITHUN ET AL.5
Overall, the amount of fatty acids in the treated fish increased compared to the control. The smallest increase
was observed in saturated fatty acids whereas the monounsaturated fatty acids showed a significant increase in the
level expressed (Table 7). There was only a marginal increase in polyunsaturated fatty acid levels in the treated fish.
3.4 |Fish growth
No statistically significant differences were observed in growth or survival (Table 8) in response to the different pro-
tein and lipid concentration in the feeds (p> .05). However, the highest growth and SGR levels were recorded for
feed containing 40% protein and 9% lipid. Therefore, those fed with 40% protein and 9% lipid may show relatively
TABLE 3 Proximate chemical composition of experimental feeds and commercial feed used as experimental diets
Analyzed composition
(% on dry matter basis) 306 309 406 409 506 509 Commercial feed
CP 30.78 31.02 40.16 40.92 50.33 49.98 51.00
EE 6.46 9.53 6.28 9.36 6.62 9.25 11.00
CF 1.30 1.22 0.87 2.02 1.37 1.34 3.50
NFE 54.1 50.97 42.77 38.03 29.24 27.00 23.50
Ash 7.36 7.26 9.92 9.67 12.44 12.43 15.00
AIA 0.31 0.33 0.49 0.65 0.77 0.88 NR
Gross energy (MJ/kg)
1
18.41 19.14 18.39 18.95 18.36 18.94 19.25
P/E ratio (mg/kJ 1.67 1.62 2.18 2.16 2.74 2.64 2.65
Note. AIA: acid-insoluble ash; CF: crude fiber; CP: crude protein; EE: ether extract or crude fat; NFE: nitrogen-free extract;
NR: not reported by the manufacturer.
1
Calculated based on Cuzon and Guillaume (1997): 21.3, 17.2, and 39.5 MJ/kg protein, carbohydrate, and lipid, respectively.
TABLE 4 Amino acid composition of experimental feeds administered during the experimental duration of 9 weeks
(g/100 g protein)
Amino
acids 306 309 406 409 506 509
Asp 4.26 0.26
a
5.65 0.23
b
4.71 0.17
ac
5.25 0.04
bc
6.22 0.11
d
3.44 0.17
e
Glu 11.44 0.11
a
12.62 0.26
b
9.53 0.26
c
9.52 0.16
c
9.26 0.16
cd
8.61 0.26
d
Ser 4.57 0.4
abc
5.36 0.4
b
4.4 0.14
a
4.5 0.06
a
4.55 0.15
a
4.72 0.11
ab
Gly 8.24 0.16
a
8.11 0.11
a
9.69 0.26
b
9.62 0.4
b
9.95 0.01
b
9.65 0.08
b
His 1.9 0.04
a
1.98 0.01
a
1.77 0.21
a
1.93 0.06
a
1.71 0.32
a
1.52 0.31
a
Arg 5.24 0.09
a
5.36 0.12
a
5.35 0.05
a
5.44 0.13
a
5.59 0.21
a
6.56 0.37
b
Thr 3.42 0.08
a
3.47 0.17
a
3.74 0.03
a
3.69 0.17
a
3.78 0.01
a
3.63 0.43
a
Ala 6.57 0.09
a
6.57 0.02
a
7.18 0.11
b
7.47 0.36
b
7.45 0.21
b
7.71 0.04
b
Pro 8.84 0.05
a
7.27 0.34
b
5.75 0.34
c
4.82 0.05
de
4.4 0.21
d
5.53 0.32
ce
Tyr 1.82 0.08
ac
1.69 0.35
a
2.17 0.05
abc
2.26 0.05
bc
2.55 0.22
b
2.45 0.05
b
Val 5.84 0.05
a
5.80 0.08
a
5.82 0.12
a
5.85 0.04
a
5.71 0.05
a
6.26 0.06
b
Met 0.87 0.15
a
0.55 0.05
b
0.89 0.05
a
0.83 0.05
a
1.25 0.06
c
0.86 0.06
a
Cys 0.24 0.02
a
0.22 0.02
a
0.19 0.12
a
0.17 0.05
a
0.30 0.01
a
0.37 0.16
a
Ile 5.71 0.11
a
5.79 0.09
a
5.92 0.04
ab
6.11 0.1
ab
5.81 0.1
a
6.36 0.36
b
Leu 8.32 0.34
a
7.58 0.24
b
8.31 0.05
a
8.26 0.04
a
8.17 0.16
ab
8.34 0.04
a
Phe 4.65 0.22
a
4.43 0.03
a
4.53 0.03
a
4.6 0.06
a
4.31 0.13
a
4.62 0.11
a
Lys 11.23 0.04
a
11.24 0.06
a
12.74 0.11
b
12.63 0.05
b
12.79 0.19
b
13.47 0.03
c
Note. All samples were analyzed in triplicate (n= 3) and expressed as means SD. Means followed by different superscript
letters within the same row are significantly different (p< .05).
6MITHUN ET AL.
TABLE 5 Fatty acid composition of experimental feeds administered during the experimental duration of 9 weeks (g/100 g fatty acids)
Fatty acids 306 309 406 409 506 509
14:0 5.17 0.07
a
5.25 0.04
a
5.52 0.03
b
5.74 0.02
c
6.13 0.06
d
7.92 0.03
e
15:0 0.34 0.02
a
0.36 0.04
a
0.49 0.05
b
0.37 0.01
a
0.53 0.02
b
0.32 0.01
a
16:0 15.33 0.12
a
13.72 0.06
b
17.79 0.02
c
17.81 0.01
c
19.54 0.04
d
27.91 0.01
e
17:0 0.53 0.05
a
0.5 0.05
a
0.8 0.01
a
0.58 0.01
a
0.4 0.51
a
0.52 0.02
a
18:0 2.56 0.03
a
2.84 0.02
b
4.23 0.02
c
4.22 0.02
c
5.32 0.01
d
5.98 0.01
e
20:0 0.86 0.04
a
0.74 0.03
b
0.28 0.01
ce
0.21 0.01
cd
0.28 0.01
e
0.22 0.02
ce
22:0 0.37 0.02
a
0.21 0.01
bd
0.18 0.02
b
0.77 0.01
c
0.36 0.03
a
0.26 0.02
d
24:0 0.19 0.02
a
0.21 0.03
a
0.06 0.02
b
0.14 0.02
ac
0.11 0.01
bc
0.17 0.02
ac
PSFA 25.36 0.14
a
23.83 0.15
b
29.35 0.13
c
29.85 0.06
c
32.68 0.46
d
43.3 0.11
e
14:1 0.14 0.04
abc
0.15 0.01
ab
0.17 0.01
b
0.11 0.01
ac
0.16 0.01
ab
0.09 0.01
c
15:1 0.15 0.01
ac
0.09 0.02
b
0.18 0.01
a
0.11 0.01
bc
0.25 0.03
d
0.14 0.01
ac
16:1 7.1 0.12
a
7.17 0.05
a
7.51 0.02
c
7.91 0.02
d
7.6 0.01
c
10.55 0.01
e
18:1n9 26.76 0.01
a
24.89 0.01
b
23.11 0.11
c
25.54 0.01
d
20.97 0.02
e
28.57 0.01
f
20:1 0.35 0.02
a
0.22 0.02
be
0.28 0.01
c
0.54 0.01
d
0.25 0.03
ce
0.15 0.01
f
22:1 0.18 0.01
ac
0.22 0.02
a
0.18 0.02a
c
0.97 0.01
b
0.17 0.01
c
0.09 0.01
d
24:1 0.55 0.01
a
0.11 0.01
b
0.65 0.01
c
0.58 0.01
a
0.83 0.02
d
0.06 0.01
b
PMUFA 35.22 0.19
a
32.84 0.05
b
32.08 0.17
c
35.75 0.05
d
30.24 0.08
e
39.65 0.06
f
18:2n6 22.73 0.02
a
19.07 0.01
b
17.14 0.03
c
18.87 0.02
d
14.57 0.01
e
14.26 0.02
f
18:3n6 2.27 0.01
ac
2.18 0.01
b
2.22 0.02b
c
1.54 0.02
d
1.93 0.04
e
0.43 0.01
f
18:3n3 1.52 0.03
a
1.52 0.02
a
0.28 0.01
b
0.28 0.02
b
0.17 0.01
c
0.14 0.03
c
20:2n6 1.26 0.02
a
1.19 0.01
b
1.36 0.03
c
1.15 0.03
b
0.91 0.02
d
0.55 0.03
e
20:3n6 0.66 0.02
a
0.78 0.02
b
0.91 0.01
c
0.96 0.02
d
1.22 0.02
e
0.21 0.01
f
20:4n6 (ARA) 0.19 0.01
a
0.55 0.01
b
0.43 0.01
c
0.77 0.01
d
0.54 0.02
b
0.28 0.01
e
20:5n3 (EPA) 5.34 0.03
a
6.55 0.03
b
6.04 0.04
c
6.35 0.01
d
5.98 0.02
e
0.96 0.02
e
22:6n3 (DHA) 2.05 0.02
a
4.07 0.01
b
4.54 0.01
c
3.44 0.04
d
4.80 0.00
e
0.31 0.02
f
PPUFA 36.02 0.11
a
35.9 0.09
a
32.92 0.09
b
33.36 0.08
e
30.11 0.09
d
17.15 0.12
e
Other (unidentified) 3.8 0.05
a
6.87 0.07
b
6.14 0.05
c
1.26 0.02
d
6.69 0.18
b
00.21 0.02
e
Note. All samples were analyzed in triplicate (n= 3) and expressed as means SD. Means followed by different superscript letters within the same row are significantly different (p< .05).
PMUFA: total monounsaturated fatty acids; PPUFA: total polyunsaturated fatty acids; PSFA: total saturated fatty acids; ARA: arachidonic acid; DHA: docosahexaenoic acid; EPA: eico-
sapentaenoic acid.
MITHUN ET AL.7
TABLE 6 Initial and final whole-body amino acid compositions of experimental fish (g 100/g protein)
Amino acids Initial 306 309 406 409 506 509 Commercial feed
Asp 1.61 0.02
ad
1.48 0.02
bf
1.53 0.02
bcf
1.58 0.02
ac
1.56 0.02
cd
1.67 0.03
a
1.65 0.04
ae
1.55 0.02
adf
Glu 2.53 0.02
a
2.14 0.01
b
2.2 0.01
c
2.27 0.03
d
2.27 0.01
d
2.4 0.01
e
2.37 0.01
e
2.26 0.02
d
Ser 0.77 0.02
acd
0.69 0.01
be
0.69 0.01
be
0.73 0.02
abd
0.69 0.03
be
0.79 0.01
c
0.76 0.02
cd
0.67 0.02
e
Gly 1.54 0.01
a
1.51 0.01
ad
1.35 0.01
b
1.66 0.03
c
1.46 0.02
d
1.66 0.01
c
1.62 0.02
c
1.49 0.03
ad
His 0.43 0.02
a
0.35 0.01
b
0.36 0.01
b
0.38 0.02
ab
0.37 0.01
b
0.38 0.01
ab
0.37 0.01
b
0.34 0.02
b
Arg 1.14 0.02
a
0.98 0.01
bcd
0.95 0.01
c
1.05 0.02
bde
0.98 0.01
cdf
1.08 0.01
ae
1.08 0.01
ae
1.03 0.04
bef
Thr 0.79 0.01
a
0.65 0.01
b
0.68 0.01
bd
0.69 0.03
bcd
0.68 0.01
bd
0.74 0.01
ac
0.74 0.02
ad
0.67 0.01
b
Ala 1.24 0.02
acd
1.15 0.01
b
1.14 0.02
b
1.23 0.03
dc
1.17 0.02
bc
1.3 0.02
d
1.27 0.03
d
1.18 0.01
b
Pro 1.15 0.06
a
0.56 0.01
b
0.56 0.01
b
0.6 0.02
bc
0.64 0.01
bc
0.68 0.01
c
0.68 0.02
c
0.57 0.01
b
Tyr 0.54 0.01
a
0.41 0.01
b
0.44 0.02
bc
0.43 0.01
bc
0.43 0.01
bc
0.46 0.01
c
0.43 0.02
bc
0.43 0.01
bc
Val 0.84 0.04
a
0.83 0.04
a
0.74 0.02
b
0.88 0.01
ac
0.93 0.02
c
0.88 0.01
ac
0.93 0.01
c
0.91 0.01
ac
Met 0.45 0.03
a
0.41 0.02
ab
0.39 0.01
ab
0.42 0.02
ab
0.42 0.03
ab
0.43 0.01
a
0.42 0.01
ab
0.36 0.02
b
Cys 0.14 0.02
a
0.33 0.02
bc
0.33 0.02
b
0.36 0.01
bc
0.35 0.01
bc
0.38 0.01
cd
0.43 0.01
d
0.35 0.01
bc
Ile 0.68 0.01
a
0.55 0.01
bc
0.53 0.01
b
0.59 0.03
bc
0.64 0.01
ad
0.6 0.01
cd
0.64 0.02
ad
0.65 0.02
ad
Leu 1.23 0.02
a
1.03 0.02
b
1.05 0.01
b
1.12 0.02
c
1.13 0.01
c
1.17 0.01
c
1.15 0.02
c
1.12 0.02
c
Phe 0.75 0.03
a
0.58 0.01
b
0.6 0.01
bc
0.62 0.01
bc
0.63 0.02
bc
0.66 0.02
c
0.64 0.02
bc
0.62 0.01
bc
Lys 1.24 0.02
a
1.79 0.02
bc
1.74 0.02
b
1.84 0.01
cde
1.85 0.01
de
1.90 0.01
e
1.84 0.02
cd
1.78 0.03
bef
Note. All samples were analyzed in triplicate (n= 3) and expressed as means SD. Means followed by different superscript letters within the same row are significantly different (p< .05).
8MITHUN ET AL.
TABLE 7 Initial and final fatty acid compositions of experimental fish (g 100/g lipid)
Fatty acids Initial 306 309 406 409 506 509 Commercial feed
12:0 0.04 0.02
a
0.03 0.02
a
0.02 0.01
a
0.03 0.02
a
0.03 0.01
a
0.03 0.01
a
0.02 0.01
a
0.02 0.01
a
14:0 0.22 0.02
a
0.25 0.01
ab
0.28 0.01
bc
0.26 0.02
ab
0.29 0.01
bc
0.27 0.01
bc
0.31 0.01
c
0.48 0.01
d
16:0 1.46 0.02
a
2.54 0.01
bf
2.51 0.01
bf
2.43 0.03
c
2.49 0.01
bcd
2.49 0.02
bd
2.52 0.02
bdf
2.55 0.01
f
18:0 0.37 0.01
a
0.47 0.01
b
0.46 0.02
b
0.43 0.03
ab
0.47 0.03
b
0.47 0.02
b
0.46 0.02
b
0.46 0.01
b
PSFA 2.1 0.05
a
3.3 0.05
b
3.26 0.03
b
3.15 0.09
b
3.27 0.05
b
3.26 0.05
b
3.31 0.05
b
3.52 0.04
c
16:1n7 0.67 0.02
a
0.98 0.02
b
0.76 0.02
a
1.02 0.03
bc
1.04 0.06
bc
1.07 0.01
c
1.07 0.02
c
0.88 0.01
d
18:1n9 1.27 0.01
a
2.64 0.04
b
2.61 0.01
b
2.35 0.04
c
2.4 0.01
c
2.19 0.03
d
2.33 0.04
c
1.78 0.01
e
PMUFA 1.94 0.03
a
3.62 0.03
b
3.37 0.03
cd
3.37 0.05
cd
3.45 0.07
c
3.26 0.03
d
3.4 0.05
cd
2.66 0.02
e
18:2n6 0.52 0.02
a
1.06 0.01
b
1.17 0.02
c
0.89 0.01
df
0.92 0.01
d
0.76 0.01
e
0.86 0.01
f
0.52 0.02
a
18:3n3 0.14 0.01
a
0.09 0.01
b
0.08 0.01
b
0.08 0.01
b
0.08 0.01
b
0.08 0.01
b
0.08 0.01
b
0.08 0.01
b
20:2n6 0.06 0.01
a
0.06 0.01
a
0.05 0.01
a
0.03 0.01
a
0.05 0.01
a
0.06 0.01
a
0.05 0.02
a
0.11 0.01
b
20:4n6 (ARA) 0.11 0.01
a
0.08 0.01
ab
0.07 0.01
b
0.08 0.01
ab
0.07 0.01
ab
0.10 0.01a
b
0.10 0.01
ab
0.08 0.01
ab
20:5n3 (EPA) 0.13 0.02
a
0.07 0.02
b
0.08 0.02
b
0.07 0.01
b
0.07 0.02
b
0.08 0.01
b
0.09 0.01
ab
0.23 0.01
c
22:6n3 (DHA) 0.44 0.02
a
0.22 0.01
bd
0.19 0.01
bd
0.23 0.04
bd
0.17 0.01
bc
0.23 0.01
bd
0.24 0.01
d
0.45 0.01
a
PPUFA 1.58 0.06
acd
1.71 0.07
abcd
1.88 0.08
bc
1.92 0.08
b
1.66 0.07
cd
1.56 0.03
d
1.62 0.07
d
1.71 0.06
abcd
Note. All samples were analyzed in triplicate (n= 3) and expressed as means SD. Means followed by different superscript letters within the same row are significantly different (p< .05).
PMUFA: total monounsaturated fatty acids; PPUFA: total polyunsaturated fatty acids; PSFA: total saturated fatty acids; ARA: arachidonic acid; DHA: docosahexaenoic acid; EPA: eico-
sapentaenoic acid.
MITHUN ET AL.9
higher growth rates than the control. There were no significant differences among treatments in terms of survival
rate (p> .05).
3.5 |Fish maturation and breeding
After 63 days of feeding trial, spawning was observed in the group receiving treatment containing 40% protein and
6% lipid 7 days later. Spawning was also observed in the other two treatment groups receiving 50% protein and 6%
lipid and 50% protein and 9% lipid after 84 and 90 days of feeding, respectively. The commercial feed treatment
group spawned after 103 days of feeding. No other spawning was observed for the remaining treatment groups even
after 120 days of feeding.
3.6 |Whole-body composition
Whole-body composition of honey gourami is shown in Table 9. Significant differences (p< .05) in protein and lipid
content were observed after the feeding trial. There was a significant decrease in both crude protein (CP) and crude
lipid (EE) relative to the initial levels. Total ash content also showed significant difference from the initial level but
was comparatively lower than that shown for both the protein and lipid levels.
4|DISCUSSION
Growth and survival did not significantly differ among honey gourami, T. chuna, that were fed on feeds containing
various levels of dietary protein (30, 40, and 50%) and lipid (6 and 9%). Shim et al. (1989) reported that 35% dietary
protein in the diet resulted in increased growth rates and percentage of vitellogenic oocytes in T. lalius. Degani and
Gur (1992) evaluated the gross protein requirements of pearl gourami, T. leerii, using formulated feeds containing
protein levels ranging from 13% to 49%. They observed faster growth in fish receiving feeds containing 1332% pro-
tein level than those receiving feeds containing 3249% protein. As such, it was concluded that 2632% was the
optimal dietary protein range for this species.
Improvement in the growth rate by about 67% was observed in fish receiving feed 409 (40% protein and 9%
fat). Mohanta and Subramanian (2007) fed blue gourami, Trichogaster trichopterus, semipurified diets containing
casein, gelatin, and dextrin and were able to demonstrate that a feed composition of 35% protein, 8% fat, and 40%
carbohydrate, with a digestible energy (DE) level of 16.7 MJ/kg, was optimal for growth and nutrient utilization for
this species. Therefore, the suitable feed composition containing 30% protein and 6% lipid levels would be sufficient
for regular maintenance of these fish in an aquarium setting.
In the present study, amino acid levels of the experimental diets were higher than those for the whole body both
before and after the feeding trial. Ogino (1980) reported that gross body composition and feed amino acid profiles
TABLE 8 Effect of different experimental treatments on the growth and survival of experimental fish (Trichogaster
chuna)
Treatments 306 309 406 409 506 509
Commercial
feed
Initial weight (g) 1.34 0.04 1.36 0.05 1.34 0.03 1.35 0.05 1.35 0.03 1.38 0.03 1.38 0.01
Final weight (g) 2.16 0.07 2.12 0.22 2.22 0.12 2.26 0.04 2.19 0.07 2.25 0.05 2.24 0.12
Weight gain (g) 0.82 0.04 0.76 0.18 0.87 0.09 0.90 0.02 0.83 0.05 0.86 0.08 0.85 0.11
Weight gain (%) 61.09 1.97 54.96 11.51 64.67 5.35 67.28 4.20 61.63 3.49 62.90 6.56 61.50 7.77
SGR
1
0.76 0.03 0.72 0.11 0.78 0.06 0.81 0.02 0.77 0.03 0.80 0.02 0.79 0.05
Survival (%) 97.50 2.04 92.50 6.24 99.16 0.68 97.50 1.18 96.33 0.95 95.83 1.80 92.50 2.04
1
Specific growth rate (SGR) = (L
n
[final weight] L
n
[initial weight])/time, where L
n
is natural logarithm.
10 MITHUN ET AL.
TABLE 9 Initial and final whole-body biochemical composition of the experimental fish (%)
Treatments
Initial composition 306 309 406 409 506 509 Commercial feed
CP 19.25 0.04
a
17.25 0.03
b
17.56 0.04
c
17.68 0.02
d
16.3 0.01
e
17.06 0.03
f
16.57 0.01
g
17.24 0.04
b
EE 8.03 0.02
a
10.86 0.02
b
10.34 0.02
c
9.82 0.02
d
12.05 0.01
e
11.07 0.05
f
11.34 0.02
g
10.16 0.02
h
CF 0.25 0.03
a
0.08 0.02
bc
0.05 0.02
c
0.1 0.01
bc
0.23 0.07
a
0.16 0.02
ab
0.06 0.01
bc
0.07 0.01
bc
NFE 0.17 0.02
a
0.14 0.01
a
0.07 0.02
b
0.24 0.02
c
0.03 0.02
b
0.3 0.02
c
1.1 0.01
d
0.16 0.02
a
Ash 8.62 0.02
a
7.96 0.01
b
8.14 0.02
c
8.42 0.02
d
7.7 0.01
e
7.61 0.06
e
7.2 0.01
f
8.59 0.03
a
Note. All samples were analyzed in triplicate (n= 3) and expressed as mean SD. Means followed by different superscript letters within the same row are significantly different (p< .05).
CF: crude fiber; CP: crude protein; EE: ether extract; NFE: nitrogen-free extract.
MITHUN ET AL.11
are important indicators of nutrient requirements because the ideal protein level (Emmert & Baker, 1997; Wang &
Fuller, 1989) in the feed should reflect the whole-body essential amino acid profile. The amino acid composition of
feeds was higher than that of the whole body (Tables 4 and 6). Therefore, the amino acid oxidation rate could have
been low, and the amount of amino acids available exceeded that required for protein synthesis (Kaushik & Seiliez,
2010). Dietary protein in excess of the required level was deaminated and used in lipid synthesis (Bai, Wang, & Cho,
1999; Shah Alam, Watanabe, & Carroll, 2008; Siddiqui & Khan, 2009; Zehra & Khan, 2012). This phenomenon was
evident in the present study and indicated an increase of about 4% in the final body lipid levels compared to those
that were measured initially (Table 9).
The amino acid requirements of fish may be related to body composition (Mambrini & Kaushik, 1995). However,
it remains unclear why only whole-body lysine and cysteine levels were higher at the end of the feeding trial than
they were initially in the present study. The effect of dietary amino acids on amino acid utilization is poorly under-
stood and remains controversial (Bureau & Encarnação, 2006; Encarnação et al., 2004, 2006). However, National
Research Council (2011) suggested three approaches that could meet amino acid requirements, namely, an increase
in dietary protein levels, supplementation with crystalline amino acids, and combining various protein sources with
different amino acid profiles. The third approach was adopted in the present study. Only lysine and cysteine levels
could be correlated between the amino acid compositions of the feed and the whole body. Lysine is considered the
least toxic of the amino acids (Sauberlich, 1961). However, excess lysine may impede fish growth (Bicudo, Sado, &
Cyrino, 2009; Mai et al., 2006). In the present study, no such effect was confirmed because the fish exhibited sexual
maturation, which would not have occurred if there had been growth depression. However, the relative differences
in the fish growth performance seen in the treatments were not significant. In general, methionine and cysteine
requirements are considered together. If cysteine could successfully replace methionine, then the dietary levels of
the latter could be reduced (Farhat & Khan, 2014; Poppi, Moore, & Glencross, 2017). In the present study, it was
inferred that the methionine levels in the feed were adequate based on the observation made when there was an
increase in the levels of tissue cysteine.
The proximate body composition of fish varies in response to certain endogenous and exogenous factors. Lipid
level is dependent on dietary lipid content and quality. Vegetable oils had been reported to show a positive effect on
anabantids (Berenjestanaki et al., 2014). In the present study, groundnut oil was blended with crude sardine oil at a 1:1
ratio and used in the experimental diets. This mixture yielded better results than using either type of oil alone. Using
similar diets, Tocher (2003) observed that excessive lipid was generally deposited in the visceral cavity and liver. How-
ever, the present study showed a gross body lipid increase of only 4% in T. chuna (Table 9). This disparity could be
because of the comparatively small size of the fish used in this study. Once the threshold lipid level exceeds the normal
level, small fish could dispose of the excess fat rather than letting it accumulate in the body. This theory was substanti-
ated by the changes observed in the whole-body fatty acid profiles (Table 7). A two-fold increase was observed only in
the monounsaturated fatty acid fraction. The relative increases in the saturated and polyunsaturated lipid fractions were
only marginal. A significant increase in whole-body polyunsaturated lipid concentration was reported by Wang
et al. (2014). Nevertheless, these results were inconsistent with those reported in the present study. Most of the previ-
ous studies addressing this topic focused on food fish rather than ornamental fish. Further investigation needs to be
conducted to elucidate these dynamics as observed in the gourami used in the study. Fish receiving 40% protein and
6% lipid (406) matured sexually and spawned in 7 weeks whereas those that received higher protein and lipid levels in
their diet spawned 57 weeks later (506:12 weeks; 509:13 weeks and commercial feed of 51% protein and 11% fat:
14 weeks). Tissue lipid levels (Table 9) showed an increase from 8.03% to 9.82% with treatment 406 and a greater
increase of 12.05% with treatment 409. Fish with the lowest tissue lipid levels matured and spawned first. The ratios of
DHA:EPA:ARA were 2:3:0.2 in the feeds and 0.23:0.08:0.10 in the tissues. The significance of such transformations
would require further investigation. In a recent study on blue gourami, Trichopodus trichopterus, Asil, Kenari, Miyanji,
and Krak (2017) suggested that maturation diets would need to be supplemented with 1% ARA. This recommendation
challenges the widely accepted hypothesis that the highly unsaturated fatty acid requirements of freshwater fish are
met by their endogenous chain elongation and desaturation systems.
12 MITHUN ET AL.
Based on the findings from present and past studies, it is inferred that 30% protein and 6% lipid levels in feed
were sufficient for T. chuna. However, in order to accelerate sexual maturation and breeding, feed containing 40%
protein and 6% lipid would be preferred. In the present study, fish receiving this type of feed spawned first, and they
grew rapidly. Zuanon et al. (2013) recently reported that feed with 37% protein was optimal for T. lalius. Compared
with other species, T. chuna is the smallest of all the gourami species, but it matures faster, although its growth is
slow. Gouramis could reach a marketable size in 120 days (Cole et al., 1999). Nevertheless, Trichogaster trichopterus
matures in 8698 days (McKinnon & Liley, 1987). The growth milestones of T. chuna were reached sooner when the
marketable size was 1.3 g. Sexual maturation was observed when the fish reached 2.2 g.
In conclusion, 30% protein and 6% lipid levels were found to be sufficient for the normal maintenance of honey
gourami in aquaria whereas 40% protein and 6% lipid levels helped to accelerate sexual maturation and spawning.
The smallest anabantid, T. chuna, is a multibatch, asynchronous spawner, and its amino acid and fatty acid require-
ments for growth and reproduction merit further investigation.
ACKNOWLEDGMENTS
This study was financially supported by the Department of Biotechnology (DBT), Government of India, in the form of
a DBT-CREST Award (20112012) to V.P. through BT/IN/DBT-CREST Awards/46/PV/2011-12. We thank the
Director of CMFRI, DDG (Fisheries), ICAR, and DARE, Government of India, for facilitating award acceptance and
travel abroad. We thank AVA, Singapore, for their timely fatty acid and amino acid profiling of the fish samples. We
also thank Jomar Bo Lucanas for his assistance during the fish quarantine at Temasek Polytechnic.
ORCID
Pananghat Vijayagopal http://orcid.org/0000-0001-7165-2039
REFERENCES
Asil, S. M., Kenari, A. A., Miyanji, G. R., & Krak, G. V. D. (2017). The influence of dietary arachidonic acid on growth reproduc-
tive performance and fatty acid composition of ovary, egg and larvae in an anabantid model fish, blue gourami (Trichopo-
dus trichopterus; Pallas, 1770). Aquaculture,476,818.
Association of Official Analytical Chemists. (1995). Official methods of analysis. Arlington, VA: Author.
Association of Official Analytical Chemists. (2005). Official method of analysis. Rockville, MD: Author.
Bai, S. C., Wang, X. J., & Cho, E. S. (1999). Optimum dietary protein level for maximum growth of juvenile yellow puffers.
Fisheries Science,65, 380383.
Berenjestanaki, S., Fereidouni, A. E., Ouraji, H., & Khalili, K. J. (2014). Influence of dietary lipid sources on growth, reproduc-
tive performance and fatty acid compositions of muscle and egg in three-spot gourami (Trichopodus trichopterus) (Pallas,
1770). Aquaculture Nutrition,20, 494504. https://doi.org/10.1111/anu.12012
Bicudo, A. J. A., Sado, R. Y., & Cyrino, J. E. P. (2009). Dietary lysine requirement of juvenile pacu Piaractus mesopotamicus
(Holmberg 1887). Aquaculture,297, 151156.
Bureau, D. P., & Encarnação, P. M. (2006). Adequately determining the amino acid requirements in fish: The case example of
lysine. In L. E. Cruz-Suarez, D. Ricque-Marie, M. Tapia-Salazar, M. G. Nieto-Lopez, D. A. Villareal Cavazos, A. C. Puello
Cruz, & A. Garica-Ortega (Eds.), Avances en Nuticion Acuicola, VIII Simposim International de Nutricion Acuicola
(pp. 2954). Mazatlan, Mexico: Universidad Autonoma de Nuevo Leon.
Cole, B. M. S.,Tamaru, C. S., Bailey, R. B. A., Brown, C.(1999). A manual for commercial production of the gourami Trichogaster
trichopterus. CTSA Publication No. 35. Honolulu, HI: University of Hawaii Sea Grant College Program.
Cuzon, G., & Guillaume, J. (1997). Energy and protein: Energy ratio. In L. R. D'Abramo, D. E. Conklin, & D. M. Akiyama (Eds.),
Advances in world aquaculture-crustacean nutrition (Vol. 6, pp. 5170). Baton Rouge, LA: World Aquaculture Society.
Degani, G., & Gur, N. (1992). Growth of juvenile Trichogaster leerii (Bleeker, 1852) on diets with various protein levels. Aqua-
culture & Fisheries Management,23, 161166.
Emmert, J. L., & Baker, D. H. (1997). Use of the ideal concept for precision formulation of amino acid level in broiler diets.
Journal of Applied Poultry Research,6, 462470.
Encarnação, P. M., de Lange, C., Rodehutscord, M., Hoehler, D., Bureau, W., & Bureau, D. P. (2004). Diet digestible energy
content affects lysine utilization, but not dietary lysine requirements in rainbow trout (Oncorhyncus mykiss) for maximum
growth. Aquaculture,235, 569586.
MITHUN ET AL.13
Encarnação, P. M., de Lange, C., Rodehutscord, M., Hoehler, D., Bureau, W., & Bureau, D. P. (2006). Diet energy
source affects lysine utilization for protein deposition in rainbow trout (Onchorynchus mykiss). Aquaculture,261,
13711381.
Farhat, & Khan, M. A. (2014). Total sulfur amino acid requirement and cysteine replacement value for fingerling stinging cat-
fish, Heteropneustes fossilis (Bloch). Aquaculture,426427, 270281.
Folch, J., Lees, M., & Sloane-Stanley, G. H. (1957). A simple method for the isolation and purification of total lipids from ani-
mal tissues. Journal of Biological Chemistry,226, 497509.
Heinrikson, R. L., & Meredith, S. C. (1984). Amino acid analysis by reverse-phase high-performance liquid chromatography:
Precolumn derivatization with phenylisothiocyanate. Analytical Biochemistry,136,6574. https://doi.org/10.
1016/0003-2697(84)90307-5
Kaushik, S. J., & Seiliez, I. (2010). Protein and amino acid nutrition and metabolism in fish: Current knowledge and future
needs. Aquaculture Research,41, 322332.
Ling, K. H., & Ling, L. Y. (2006). The status of ornamental fish industry in Singapore. Singapore Journal of Primary Industry,32,
5969.
Mai, K., Zhang, L., Ai, Q., Duan, Q., Zhang, C., Li, H., Liufu, Z. (2006). Dietary lysine requirement of juvenile seabass (Lateo-
labrax japonicus). Aquaculture,258, 535542.
Mambrini, M., & Kaushik, S. J. (1995). Indispensible amino acid requirements of fish: Correspondence between quantitative
data and amino acid profiles of tissue proteins. Journal of Applied Ichthyology,11, 240247.
McKinnon, J. S., & Liley, N. R. (1987). Asymmetric species specificity in responses to female sexual pheromone by males of
two species of Trichogaster (Pisces: Belontiidae). Canadian Journal of Zoology,65, 11291134. https://doi.org/10.1139/
z87-176
Mohanta, K. N.,&Subramanian, S. (2007). Effect of protein and lipid levels on growth and nutrient utilization of blue gourami,
Trichogaster trichopterus in a closed water system. Paper presented in 8th Asian Fisheries Forum, Central Marine Fisheries
Research Institute, Kochi, India (Abstract No. PEO 023).
Mookerjee, H. K., & Majumder, S. R. (1960). On the history of Colisa lalius (Hamilton). Proceedings of the Zoological Society of
Bengal,13,2938.
Morrison, W. R., & Smith, L. M. (1964). Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron
fluoride-methanol. Journal of Lipid Research,5, 600608.
National Research Council. (2011). Nutrient requirements of fish and shrimp. Washington, DC: The National Academies Press.
Ogino, C. (1980). Requirements of carp and rainbow trout for essential amino acids. Nippon Suisan Gakkaishi,46, 171175.
https://doi.org/10.2331/suisan.46.171
Poppi, D. A., Moore, S. S., & Glencross, B. D. (2017). Redefining the requirement for total sulfur amino acids in the diet of
barramundi (Lates calcarifer) including assessment of the cysteine replacement value. Aquaculture,471, 213222.
Santamaria, V. Y., & Santamaria, C. W. (2011). Nutritional requirements of freshwater ornamental fish: A review. Revista
MVZ Cordoba,16, 24582469.
Sauberlich, H. E. (1961). Studies on the toxicity and antagonism of amino acids for weanling rats. Journal of Nutrition,75,
6172.
Shah Alam, M., Watanabe, W. O., & Carroll, P. M. (2008). Dietary protein requirements of juvenile black sea bass, Centropris-
tis striata.Journal of the World Aquaculture Society,39, 656663. https://doi.org/10.1111/j.1749-7345.2008.00204.x
Shim, K. F., Landesman, L., & Lam, T. J. (1987). Culture of the dwarf gourami Coliosa lalia in Singapore. Journal of the World
Aquaculture Society,18(3), 203206.
Shim, K. F., Landesman, L., & Lam, T. J. (1989). Effect of dietary protein on growth, ovarian development and fecundity in
the dwarf gourami, Colisa lalia (Hamilton). Journal of Aquaculture in the Tropics,4, 111123.
Siddiqui, T. Q., & Khan, M. A. (2009). Effects of dietary protein levels on growth, feed utilization, protein retention efficiency
and body composition of young Heteropneustes fossilis (Bloch). Fish Physiology and Biochemistry,35, 479488. https://
doi.org/10.1007/s10695-008-9273-7
Tocher, D. R. (2003). Metabolism and functions of lipids and fatty acids in teleost fish. Reviews in Fisheries Science,11,
107184. https://doi.org/10.1080/713610925
Vijayagopal, P. (2004). Blending of ingredients in aquafeed formulation the Excelway: Tips to farmers and managers. Fishing
Chimes,23,1723.
Vishwanath, W.(2010). Trichogaster lalius. The IUCN Red List of Threatened Species 2010:e.T166445A6210533. Retreived
from https://doi.org/10.2305/IUCN.UK.2010-4.RLTS.T166445A6210533.en
Wang, A., Yang, W., Shen, Y., Han, G., Lv, F., Yu, Y., Zhang, J. (2014). Effects of dietary lipid levels on growth performance,
whole body composition and fatty acid composition of juvenile gibel carp (Carassius auratus gibelio). Aquaculture Research,
46, 28192828. https://doi.org/10.1111/are.12571
Wang, T. C., & Fuller, M. F. (1989). The optimum dietary amino acid pattern for growing pig. 1. Experiments by amino acid
deletion. British Journal of Nutrition,62,7789.
Zehra, S., & Khan, M. (2012). Dietary protein requirement for fingerling Channa punctatus (Bloch), based on growth, feed
conversion, protein retention and biochemical composition. Aquaculture International,20, 383395.
14 MITHUN ET AL.
Zuanon, J. A. S., Carneiro, A. P. S., Nascimento, S. L. A. D., Da Silva, M. D., Pontes, M., Kanashiro, M. Y., & Salaro, A. L.
(2013). Protein requirement for Trichogaster lalius, blue variety: A short communication. Revista Brasileira de Zootecnica,
42, 144147.
How to cite this article: Mithun S, Vijayagopal P, Chakraborty K, Chan DPS. Evaluation of gross protein and
lipid requirements in formulated feed for honey gourami, Trichogaster chuna (Hamilton 1822). J World Aqua-
cult Soc. 2018;115. https://doi.org/10.1111/jwas.12557
MITHUN ET AL.15
This research hasn't been cited in any other publications.
  • Article
    This study investigates the influence of arachidonic acid (ARA) on growth, reproductive performance, and fatty acid compositions of ovary, egg and 3-DPH (days post hatching) larvae of Trichopodus trichopterus broodstock. A 150-day feeding experiment was performed comparing the effects of five ARA-supplemented diets containing 0 (control), 0.5, 1, 1.5 and 2% ARA (of total fatty acids) (TFA). Growth parameters including final weight, weight gain and specific growth rate (SGR) did not differ between different groups (P > 0.05). Feed conversion ratio (FCR) was significantly lower in control group compared to 0.5 and 1.5% ARA groups (P < 0.05). Contrary to growth indices, all reproductive traits were significantly affected by ARA contents in diets. Absolute and relative fecundities increased in relation to the ARA content (P < 0.05). The maximum absolute and relative fecundities (13.3 and 974.3 × 10³, respectively) were recorded in 2% ARA treatment group (P < 0.05). Mean oocyte diameter reached the greatest size (861.66 μm) in 1% ARA and was significantly different than other treatments (P < 0.05). The yolk sac diameter was enhanced linearly by increasing dietary ARA (P < 0.05). Results showed striking effects of dietary ARA on its accumulation in ovaries, eggs and larva. Ʃ n − 6 series fatty acid (44.9%) and its constituent (18:2n − 6) (40.85%) reached the highest contents in 0.5% ARA in ovaries (P < 0.05). A similar trend was seen in fertilized eggs and 3-DPH larvae in that the highest amounts of 18:2-6 and Σ n − 6 were found in the 0.5% ARA group (P < 0.05). A second order polynomial regression was employed for estimating ARA/EPA (eicosapentaenoic acid) ratios and showed that the respective ratio enhanced linearly in the ovary and fertilized egg. Correspondingly, ARA/DHA (docosahexaenoic acid) ratio in ovary, fertilized egg and larvae reflected those of the experimental diets. Their fatty acid profiles reflect the diet composition well, ovary (R² = 0.87), fertilized egg (R² = 0.91) and 3DPH larvae (R² = 0.82) and the ARA/DHA ratios were linearly improved by increasing dietary ARA (P < 0.05). Overall, the results of this study revealed the necessity of ARA inclusion in T. trichopterus broodstock diets. This conclusion challenges the generally accepted hypothesis that freshwater fish meet their highly unsaturated fatty acids (HUFA) requirements by elongation and desaturation their precursors. These findings showed that T. trichopterus broodstock, as a model of asynchronous multi-batch spawning fish, needs to receive at least 1% ARA in their maturation diet to improve reproductive performance. The best efficiency was achieved in 2% ARA, based on fecundity, yolk sac diameter and hatching rate.
  • Article
    This study was designed to confirm a previous estimate of the methionine (Met) and total sulfur amino acid (TSAA) requirement of juvenile barramundi (Lates calcarifer) (Coloso et al., 1999) with a view for further study. Triplicate groups of fish (initial weight: 18.3 g ± 1.5 g) were fed diets with graded levels of dietary Met (7.2–12.8 g kg− 1 DM), centred around a previously reported requirement, and a constant dietary cystine (Cys) inclusion (5.9 g kg− 1 DM) over a 42 day period. At the termination of the experiment, a significant linear increase (p < 0.001) in %BW gain was observed in response to increasing dietary methionine, with no plateau in growth, suggesting the previous estimate of requirement may have been inadequate. A second experiment was designed to re-evaluate the Met/TSAA requirement in which a broader range of methionine inclusion levels were assessed (8.6–21.4 g kg− 1 diet DM Met). Triplicate groups of fish (initial weight: 36.4 g ± 8.3 g) were fed the diets for a period of 49 days. A plateau and subsequent depression in growth, as well as significant (p < 0.05) effects of dietary Met inclusion on %BW gain, feed conversion ratio (FCR) and protein retention efficiency (PRE) were observed at the conclusion of this experiment. The best fitting of nine nutrient response models, the Compartmental Model (R² = 0.71), predicted a requirement for Met of between 10.5 (95% of maximum response) and 13.6 g kg− 1 (99% of maximum response) in a diet with 592 g kg− 1 CP and 6.6 g kg− 1 Cys (17.1–20.2 g kg− 1 TSAA; 1.8–2.3% CP Met + 1.1% CP Cys). This TSAA requirement is equivalent to 43–51% of the lysine content of the diets. The applicability of this mode of expression and its relation to the ideal protein concept is discussed as is the application of different response models to the data. The impact of dietary Met:Cys ratio was also investigated with results suggesting at least 40% of dietary Met can be replaced with Cys without significantly affecting animal performance. It was concluded that disparity in the estimates of Met and TSAA requirement between this study and that of Coloso et al. (1999) was likely the result of a combination of model choice, experimental design and mode of expression of the requirements.
  • Article
    Full-text available
    Six isocaloric diets containing 30, 40, 45, 50, 55 and 60% crude protein (CP) were fed to juvenile yellow puffer Takifugu obscyrus to determine their optimum dietary level of protein. After a 12-week feeding trial, weight gain of fish fed 50% CP diet was not significantly different from that of fish fed 55% CP diet, but significantly higher (P<0.05) than that of fish fed 30, 40, 45 and 60% CP diet. Also, a broken-line model analysis for weight gain indicated that the optimum dietary protein level was 50 ± 3.7% (mean ± SD). Fish fed 50% CP also had the highest feed efficiency and hepatosomatic index. Condition factor increased with dietary protein level. No significant differences existed in haematocrit, hemoglobin and survival rate among the dietary treatments. Therefore, these findings suggest that the optimum protein level is approximately 50% of dry diet for maximum growth in juvenile yellow puffer.
  • Article
    Ideal amino acid ratios for chicks during the early growth period (0 to 21 days) are well documented by empirical evidence, but suggested ratios for older birds are in need of confirmation. Based on best empirical estimates of lysine, SAA, and threonine requirements of broiler chicks during 0 to 21, 21 to 42, and 42 to 56 days of age, together with new knowledge of maintenance contributions to the total requirement for these amino acids, it appears that the ideal ratio of SAA and threonine to lysine may change very little as birds advance in age and weight towards a 56-day market weight. This paper presents regression equations that predict both digestible and total lysine, SAA, and threonine requirements at any age or weight between hatching and 56 days.