North American Journal of Aquaculture

© 2019 American Fisheries Society

ISSN: 1522-2055 print / 1548-8454 online
DOI: 10.1002/naaq.10124

ARTICLE

Effects of Feeding Rates on Growth, Digestive Enzyme Activity,

Serum Biochemical Parameters, and Body Composition of Juvenile,
Genetically Improved, Farmed Nile Tilapia Reared in an In-Pond
Raceway Recirculating Culture System

Yuyu Wang, Pao Xu, Zhijuan Nie, Quanjie Li, Nailin Shao, Jiancao Gao, and Gangchun Xu*
Ministry of Agriculture, Key Laboratory of Freshwater Fisheries and Germplasm Resource Utilization,
Freshwater Fisheries Research Center of Chinese Academy of Fishery Sciences, Wuxi 214081, China

Abstract
Fish culture using an in-pond raceway system (IPRS) is a new mode of freshwater aquaculture in China. This
study was carried out to assess the effect of feeding rates on growth performance, serum parameters, digestive enzyme
activity, and body composition of juvenile, genetically improved, farmed Nile Tilapia Oreochromis niloticus (GIFT)
reared in an IPRS. Fish with an initial mean body weight of 6.25 g were fed twice daily at rates of 3, 4, or 5% body
weight per day (BW/d) for 50 d. Results indicated that final fish body weight, weight gain, and specific growth rate of
GIFT improved with increasing feeding rates from 3% to 5%. Growth of GIFT fed 5% BW/d was significantly higher
than for those fed 3% BW/d; however, there was no significant difference from that of fish fed at 4% BW/d. Serum
parameters and digestive enzyme activities were not affected by feeding rates. Lipid content in muscle and whole body
increased with increasing feeding rates, and fish fed 5% BW/d had higher values compared with those fed 3% BW/d.
Based on growth and physiological and biochemical variables, it could be concluded that a feeding level of 5% BW/d
is suitable for juvenile GIFT fed twice daily reared in an IPRS.

Feeding cost is usually the major expenditure in inten- achieving the best growth and productivity and for mini-
sive fish farming. Feeding management plays a crucial role mizing feed loss and water pollution (Naylor et al. 2009;
in the effective use of feeds (Ng and Romano 2013). Feed- Lee et al. 2016). Most studies have revealed that the
ing rate is an important factor affecting feed efficiency, growth of fish increased with increasing feeding rate to an
growth, fillet quality, reproductive performance, produc- optimum level and then plateaued, and farmed fish typi-
tion costs, and feed wastage (Brett and Groves 1979; Cho cally are fed 1–5% of their body weight per day (BW/d)
et al. 2006; Bhujel et al. 2007). Feeding too much will (De Silva et al. 1986; Craig 2009; Sun et al. 2016). The
increase feed waste, which has a negative effect on growth, range of the feeding rates of fish is affected by differences
profit, and the environment (Cho and Bureau 2001; in life stage, time of day, season, and water quality vari-
Rowland et al. 2005), while feeding too little results in ables (e.g., water temperature, dissolved oxygen level)
poor growth, cannibalism, and greater size heterogeneity (Craig 2009).
(Ribeiro et al. 2015). Moreover, underfeeding or overfeed- In China, aquaculture has expanded rapidly over the
ing can increase the susceptibility of fish to stress and dis- past few decades. Pond culture is the most important
ease-causing agents, which could ultimately represent an freshwater aquaculture model, and it was estimated to
economic loss (Lim et al. 2015). Therefore, it is crucial to account for about 60–70% of total freshwater aquaculture
understand the appropriate feeding rate of fish for production (Li et al. 2011). However, along with the rapid

*Corresponding author: xugc@ffrc.cn

Received May 13, 2019; accepted August 18, 2019

1
2 WANG ET AL.

development of aquaculture, some issues also appeared, Feed [Zhenjiang], Jiangsu, China). Every 10 d, 20 fish in
such as genetic deterioration, disease, pollution, and sea- each raceway were weighed and the growth was estimated,
sonal oversupply (Li et al. 2011; Wang et al. 2015), and then the amount of feed was adjusted. During the trial,
these issues seriously limit the sustainable development of temperature was 24.0–31.8°C, dissolved oxygen was 3.01–
aquaculture. In China, we modified and upgraded the in- 7.18 mg/L, water flow was 0.03 m/s, pH was 7.68–8.27,
pond raceway system (IPRS) culture technology, which and ammonia-N was 0.19–0.86 mg/L. The feeding trial
was introduced by the U.S. Soybean Export Council in lasted for 50 d.
2011. In comparison with traditional static fishponds, this
new culture model has the advantage of easy management Sample Collection
and increases production efficiency while decreasing water After they were starved for 24 h, 10 fish were sampled
and chemical pollution. So far, the field mainly focuses on from each raceway for chemical analysis. Twenty fish were
the engineering design of an IPRS, stocking density, and randomly captured from each raceway and anesthetized
flesh quality (Harimana et al. 2018; Wang et al. 2019). with MS-222 (tricaine methanesulfonate; 200 mg/L). Body
However, there are no published data on feeding rate. length and body weight were measured to estimate growth
Nile Tilapia Oreochromis niloticus is an important and calculate condition factor. Blood samples were
farmed species in China because of its wide-ranging food obtained from the caudal vein using a 2-ml one-off aseptic
habits, high growth rate, high disease resistance, conve- injector. Blood was kept a centrifuge tube, centrifuged at
nient processing, short culture period, and suitability for 4,000 × g (4°C, 15 min), then the serum was separated and
high-density culture (Xu and Ming 2018). Knowledge of stored at −80°C for serum parameter analyses. Viscera
the appropriate feeding management is crucial for high- and liver were dissected and weighed to calculate viscero-
yield, high-efficiency, and good-quality Nile Tilapia farm- somatic index (VSI) and hepatosomatic index (HSI),
ing. Nevertheless, there are no published records on the respectively. Liver, intestine, stomach, and muscle were
effect of feeding rate for fish cultured in an IPRS yet. In dissected and stored at −80°C for further analysis.
practical production, fish were fed by overfeeding or ad li-
bitum; however, this may cause feed waste and water pol- Analytical Procedures
lution. Therefore, this study was conducted to evaluate the Serum biochemistry analyses.— Serum aspartate amino-
effect of feeding rates on growth performance, serum transferase (AST), alanine aminotransferase (ALT), alkaline
parameters, digestive enzyme activity, and body composi- phosphatase (ALP), total protein, triglyceride, cholesterol,
tion of juvenile, genetically improved, farmed Nile Tilapia and glucose were measured using reagent kits (Shenzhen
(GIFT) cultured in an IPRS. These data are very neces- Mindray Bio Medical Company, Shenzhen, China) with an
sary for optimizing culture conditions and feeding man- automated biochemistry analyzer (Mindray BS-400; Shen-
agement strategy in IPRS culture operations. zhan, China). Serum lysozyme, cortisol, and superoxide dis-
mutase (SOD) content were determined using an ELISA Kit
(BPRO; Langton Biotechnology Company, Shanghai,
METHODS China) with a labsystems Multiskan MS (352, Finland).
Digestive enzyme activity.— Stomach and intestinal
Feeding Experiments samples were weighed and homogenized in ice-cold 0.86%
This feeding experiment was carried out in IPRSs as sterile normal saline solution (tissue : saline of 1:9) using a
describe by Wang et al. (2019) at Yangzhong experimental high-speed tissue homogenizer. Then, the homogenates
base, Freshwater Fisheries Research Center, Chinese were centrifuged at 3,500 × g for 15 min at 4°C and the
Academy of Fishery Sciences, Jiangsu, China. resultant supernatants were collected and stored at −80°C
Juvenile GIFT were obtained from a tilapia farm of for further analysis. The amylase, trypsin, lipase, and pep-
the Freshwater Fisheries Research Center of the Chinese sin activities were measured using assay kits (Jiancheng
Academy of Fishery Sciences (Yixing, China). Before the Bioengineering Institute, Nanjing, China), according to
feeding trial, fish were reared in ponds to acclimatize them the manufacturer's instructions.
to the experimental conditions for 2 weeks. At the begin- Proximate body composition.— Crude protein was ana-
ning of this trial, fish were starved for 24 h before weigh- lyzed by the Kjeldahl method using an Auto Kjeldahl Sys-
ing. Juvenile GIFT with an average body weight of tem (K1100 Auto analyzer; Hanon Instruments, Jinan,
6.25 ± 0.32 g (mean ± SD) were stocked into six 220-m3 China). Crude lipid was analyzed using a Soxtec System
raceways at a density of 0.57 kg/m3. Fish were hand feed (Sox500; Hanon Instruments, Jinan, China). Dry matter
at a rate of 3, 4, or 5% BW/d twice daily (0700 and 1700 was analyzed by drying to a constant weight at 105°C for
hours), with two replications per treatment. Fish were fed 12 h. Ash was analyzed by combustion in a muffle furnace
with a commercial floating formulated diet, which con- at 550°C for 4 h. The fatty acid profiles of muscle samples
tained ≥28% crude protein and ≥4% crude lipids (Cargill were analyzed with a chromatograph (GC-2010; Shimadzu
 FEEDING RATES OF GENETICALLY IMPROVED FARMED NILE TILAPIA 3

Corporation, Kyoto, Japan), following procedures described fed various feeding rates ranged from 91.07% to 94.56%
by Harimana et al. (2018). (P > 0.05). Weight gain, SGR, and final body weight of
GIFT increased with increasing feeding rates from 3% to
Calculations and Statistical Analyses 5%. Fish fed 5% BW/d had significantly higher weight
The following equations were used, where IBW is the gain, SGR, and final body weight than fish fed 3% BW/d
initial body weight and FBW is the final body weight: (P < 0.05), but there was no significant difference from
those fed 4% BW/d (P > 0.05). Fish fed 3% BW/d had a
Specific growth rate ðSGR; %=dÞ ¼ ðln FBW  ln IBWÞ higher feed efficiency value than those fed 5% BW/d (P <
 100=culturing period 0.05) but were not significantly different from those fed
4% BW/d (P > 0.05). No significant differences were found
for condition factor and HSI among all treatments (P >
Weight gain ð%Þ ¼ 100  ðFBW  IBWÞ=IBW 0.05); however, VSI increased with increasing feeding rates
(P < 0.05) (Table 1).
Feed efficiency ¼ weight gain (g)=feed consumed (g)
Serum Biochemistry Analyses
No significant differences were observed in serum AST,
Condition factor ðg=cmÞ ¼ 100  ðbody weight [g]Þ ALT, ALP, total protein, glucose, cholesterol, triglyceride,
=ðbody length [cm]Þ3 cortisol, SOD, and lysozyme content among fish fed vari-
ous feeding rates (P > 0.05) (Table 2).

VSI ð%Þ ¼ 100  ðviscera weight=body weightÞ Digestive Enzyme Activity

There were no significant differences in digestive
HSI ð%Þ ¼ 100  ðliver weight=body weightÞ enzyme activity (amylase, lipase, trypsin, and pepsin)
among fish fed various feeding rates (P > 0.05) (Table 3).

The data were expressed as mean ± SD. All data from Proximate Body Composition
different treatments were subjected to one-way analysis of Muscle and whole body composition of GIFT are pre-
variance, followed by Duncan's multiple range test for sig- sented in Table 4. The feeding rates did not affect crude
nificant differences (P < 0.05). The SAS 9.12 program protein content in the whole body (P > 0.05). Fish fed 5%
(Statistical Analysis System Institute, Cary, North Caro- BW/d exhibited lower moisture and ash levels in the whole
lina) was used for statistical analysis. body compared with fish fed 3% BW/d, but no significant
differences were found between fish fed 3% and 4% BW/d
or between 4% and 5% BW/d (P < 0.05). Lipid content in
RESULTS muscle and whole body increased with increasing feeding
rates, and fish fed 5% BW/d had significantly higher lipid
Growth content than those fed 3% BW/d (P < 0.05). No significant
The growth performance of GIFT fed different feeding differences were found for moisture or protein content in
rates is presented in Table 1. Mean survival rates of fish muscle among all treatments (P > 0.05).

TABLE 1. Effects of feeding rate on the growth and somatic indexes of juvenile, genetically improved, farmed Nile Tilapia. Within a row, different
letters indicate significant differences (P < 0.05). Data are presented as mean ± SD (n = 2).

Feeding rate (% BW/d)

Measurement 3 4 5
Initial body weight (g) 6.25 ± 0.32 6.25 ± 0.32 6.25 ± 0.32
Survival rate (%) 92.38 ± 1.72 94.56 ± 0.29 91.07 ± 2.23
Final body weight (g) 92.71 ± 3.83 y 108.27 ± 5.45 zy 115.04 ± 4.78 z
Weight gain (%) 1,383.31 ± 61.26 y 1,632.31 ± 123.23 zy 1,740.63 ± 76.46 z
SGR (%/d) 5.14 ± 0.09 y 5.35 ± 0.18 zy 5.56 ± 0.08 z
Feed efficiency 0.67 ± 0.01 z 0.60 ± 0.02 zy 0.58 ± 0.01 y
Condition factor (g/cm3) 4.13 ± 0.12 4.30 ± 0.08 4.22 ± 0.14
HSI (%) 2.28 ± 0.35 2.51 ± 0.57 2.59 ± 0.52
VSI (%) 15.95 ± 0.56 y 16.45 ± 0.38 zy 17.38 ± 0.29 z
4 WANG ET AL.

TABLE 2. Effects of feeding rate on the serum biochemical parameters of juvenile, genetically improved, farmed Nile Tilapia. There were no signifi-
cant differences between any of the parameters (P < 0.05). Data are presented as mean ± SD (n = 2). Abbreviations are as follows: ALT = alanine
aminotransferase, AST = aspartate aminotransferase, ALP = alkaline phosphatase, and SOD = superoxide dismutase.

Feeding rate (% BW/d)

Measurement 3 4 5
ALT (U/L) 20.42 ± 1.11 20.26 ± 1.68 21.39 ± 0.93
AST (U/L) 47.34 ± 1.49 46.45 ± 4.00 43.26 ± 3.40
ALP (U/L) 19.66 ± 0.79 18.05 ± 1.36 17.73 ± 0.27
Total protein (g/L) 23.70 ± 0.88 22.46 ± 0.91 23.15 ± 0.30
Glucose (mmol/L) 4.95 ± 0.22 4.55 ± 0.49 4.84 ± 0.16
Cholesterol (mmol/L) 2.35 ± 0.18 2.08 ± 0.11 1.98 ± 0.05
Triglyceride (mmol/L) 3.20 ± 0.18 2.99 ± 0.31 3.01 ± 0.27
Cortisol (ng/L) 855.53 ± 59.24 798.62 ± 109.99 990.72 ± 80.81
SOD (ng/ml) 45.65 ± 0.83 48.44 ± 1.56 51.19 ± 1.70
Lysozyme (ng/ml) 24.52 ± 3.70 23.93 ± 3.67 25.85 ± 2.22

The percentages of the identified fatty acids in the mus- et al. 2010), and Indian Major Carp Catla catla (Pradhan
cle of GIFT fed different feeding rates are presented in et al. 2018), reported that the growth performance of fish
Table 5. No significant differences were found in total sat- improved with an increasing feeding rate up to a level but
urated fatty acids, C14:0, C15:0, C16:0, C17:0, C18:0, then further increases in the feeding rate did not result in
C20:0, C22:0, C16:1, C17:1, C18:1c, C18:2t, C18:3(n-6), an improvement in growth. In the present study, fish have
C20:1, C18:3(n-3), C20:2, C20:3(n-6), C24:1(n-9), C22: better adapted for feed and cultural conditions. The
1(n-9), C22:6(n-3), C20:5(n-3), total n-3 fatty acids, or growth of GIFT reared in an IPRS improved with
total n-6 fatty acids in muscle samples among dietary increasing feeding rates from 3% to 5% BW/d. Similarly,
treatments (P > 0.05). The proportions of C23:0, C18:2c, Rondán et al. (2004), El-Saidy and Gaber (2005), Baloi et
C20:4(n-6), and total polyunsaturated fatty acids al. (2017), and Liu et al. (2018) also found that feed con-
increased, while C18:1t and total monounsaturated fatty version ratio, growth, and production rate of fish signifi-
acids decreased as feeding rates increased (P < 0.05). cantly increased with increasing feeding rate. However,
Abidi and Khan (2014) observed that growth and protein
gain of fingerling Indian Major Carp increased with
DISCUSSION increasing feeding rates from 1% to 5% BW/d but signifi-
For successful and sustained aquaculture operations, cantly decreased in 6% and 7% BW/d groups. Huang et al.
the estimation of appropriate feeding and management is (2015) also found poor growth when fish were fed a higher
essential. Studies on most fish species, such as Florida red feeding rate. Abidi and Khan (2014) explained that the
tilapia Oreochromis sp. (Clark et al. 1990), Common Carp growth retardation in fish fed at the higher feeding rate
Cyprinus carpio (Shimeno et al. 1997), Nile Tilapia (De may be attributed to the over burden of dietary nitrogen
Silva et al. 1986), Asian Seabass Lates calcarifer (Harpaz in the body requiring its elimination, which is done at the
et al. 2005), Chinese Sucker Myxocyprinus asiaticus (Yuan cost of deposited nutrients.

TABLE 3. Effects of feeding rate on digestive enzyme activity in juvenile, genetically improved, farmed Nile Tilapia. There were no significant differ-
ences in digestive enzyme activity (P < 0.05). Data are presented as mean ± SD (n = 2).

Feeding rate (% BW/d)

Enzyme Location 3 4 5
Lipase (U/g protein) Intestine 33.39 ± 3.90 37.34 ± 5.28 40.63 ± 5.44
Stomach 70.35 ± 8.97 51.45 ± 5.18 64.52 ± 8.44
Amylase (U/mg protein) Intestine 0.59 ± 0.03 0.63 ± 0.08 0.64 ± 0.03
Stomach 0.44 ± 0.05 0.32 ± 0.03 0.41 ± 0.08
Trypsin (U/mg protein) Intestine 214.64 ± 10.33 199.95 ± 17.34 233.50 ± 17.26
Pepsin (U/mg protein) Stomach 14.74 ± 2.19 13.46 ± 3.23 16.07 ± 2.52
 FEEDING RATES OF GENETICALLY IMPROVED FARMED NILE TILAPIA 5

TABLE 4. Effects of feeding rate on the proximate composition of body and muscle of juvenile, genetically improved, farmed Nile Tilapia. Within a
row, different letters indicate significant differences (P < 0.05). Data are presented as mean ± SD (n = 2).

Feeding rate (% BW/d)

Proximate composition 3 4 5
Whole body (% wet weight basis)
Moisture 70.80 ± 2.21 z 69.99 ± 0.97 zy 68.23 ± 1.80 y
Protein 15.49 ± 1.22 16.04 ± 0.31 16.31 ± 0.85
Lipid 5.78 ± 0.32 y 6.41 ± 0.11 zy 7.25 ± 0.49 z
Ash 4.26 ± 0.13 z 4.14 ± 0.13 zy 3.71 ± 0.21 y
Muscle (% wet weight basis)
Moisture 76.12 ± 1.02 75.64 ± 0.23 74.38 ± 2.04
Protein 17.90 ± 1.07 18.19 ± 0.49 18.94 ± 1.70
Lipid 1.44 ± 0.11 y 1.73 ± 0.09 y 2.28 ± 0.18 z

In the present study, the optimal feeding rate was 5% mainly results from feed wastage (Puvanendran et al.
BW/d for GIFT reared in an IPRS, which is similar to 2003), which indicates that overfeeding would raise pro-
Liu et al. (2018), who found that the optimal feeding rate duction costs and nitrogen and phosphorus discharge.
for juvenile GIFT was 5% BW/d with a 35% protein diet. Fish serum biochemical parameters, such as enzymes,
This is higher than what was found in previous studies nutrients, and metabolites, have been used to evaluate
that indicated that 2–4% BW/d is suitable for better nutritional status, health status, and many biochemical
growth and production (De Silva et al. 1986; El-Saidy and processes in the metabolism (Öner et al. 2008; Satheeshku-
Gaber 2005; Bhujel et al. 2007). The higher feeding rate in mar et al. 2012). The activity of the two most important
this study could be ascribed to the sustained swimming amino-acid-metabolizing enzymes, ALT and AST, reflects
exercise of GIFT in an IPRS. As energy output is the intensity of fish protein and amino acid metabolism
increased through long-term moderate exercise, in order to (Ballantyne 2001). Also, ALP is a polyfunctional enzyme
maintain energy balance, an individual must increase food that acts as a transphosphorylase at alkaline pH and plays
and energy intake to match high-energy requirements a pivotal role in the resorption of sugar, formation of fatty
(Melzer et al. 2005). Moreover, the long-term exercise can acids, bone formation and cell metabolism, and metabo-
stimulate hypothalamic neuropeptide Y secretion or inhi- lism of carbohydrates, nucleic acids, nucleotides, and
bit leptin secretion (Lewis et al. 1993). phosphoproteins (Tandon and Chandra 1976; Zikic et al.
Generally, the feed utilization efficiency decreased with 2001). Likewise, serum glucose, triglycerides, cholesterol,
increasing feeding rates, and the feeding rates that allow and total protein commonly are used to detect the meta-
maximum growth performance are higher than those that bolic health of animals (Öner et al. 2008). Superoxide dis-
obtain the best feed utilization (Clark et al. 1990; Hardy mutase and lysozyme activity, as nonspecific immunity
1998; Yuan et al. 2010). Reducing the daily amount of factors, play an important role in the immune response
feed intake appears to compel fish to make the best use of and disease resistance of fish (Dong et al. 2018). Cortisol
the feed they have ingested without affecting growth rate is an important glucocorticoid hormone and is involved in
(Velázquez et al. 2006). In the present study, GIFT fed at the redistribution of energy during a stress response, and
3% BW/d had poor growth but higher feed efficiency than food deprivation may increase plasma cortisol levels
those fed 4% and 5% BW/d, indicating that 3% BW/d (Wunderink et al. 2012). In the present study, no signifi-
exceeded the maintenance ration level of juvenile GIFT. cant differences were observed in serum ALT, AST, ALP,
Studies on various cultured species, such as Chinese glucose, total protein, triglyceride, cholesterol, cortisol,
Sucker (Yuan et al. 2010), Brazilian Sardine Sardinella SOD, and lysozyme content among fish fed with different
brasiliensis (Baloi et al. 2017), and Nile Tilapia (El-Sayed feeding rates. These results indicated that the serum bio-
2002; Liu et al. 2018), also found that an increase in the chemical indices were not influenced by feeding rates. Sim-
feeding rates decreased feed efficiency and protein reten- ilar results were observed in GIFT (Liu et al. 2018) and
tion efficiency. The reduced feed utilization efficiency Olive Flounder Paralichthys olivaceus (Cho et al. 2006).
could be related to the higher rate of feed passing through However, Cho et al. (2007) found that an increased feed-
the digestive tract and decreased feed digestibility and ing rate led to an increase in the serum triglyceride and
metabolism (Henken et al. 1985). The increased feed con- AST of subadult Olive Flounder but had no significant
version ratio with increasing feeding rates beyond the best effect on serum total protein, glucose, and ALT. Shimeno
6 WANG ET AL.

TABLE 5. Effects of feeding rate on the fatty acid composition of muscle of juvenile, genetically improved, farmed Nile Tilapia. Within a row, differ-
ent letters indicate significant differences (P < 0.05). Abbreviations are as follows: SFA = saturated fatty acid, MUFA = monounsaturated fatty acid,
and PUFA = polyunsaturated fatty acid.

Feeding rate (% BW/d)

Fatty acid 3 4 5
C14:0 2.59 ± 0.23 2.52 ± 0.01 2.53 ± 0.02
C15:0 0.26 ± 0.03 0.32 ± 0.04 0.28 ± 0.03
C16:0 23.65 ± 0.45 22.76 ± 0.11 23.60 ± 0.59
C17:0 0.36 ± 0.04 0.43 ± 0.06 0.38 ± 0.06
C18:0 6.55 ± 0.32 6.27 ± 0.08 7.01 ± 0.43
C20:0 0.62 ± 0.02 0.61 ± 0.06 0.64 ± 0.04
C22:0 0.71 ± 0.03 0.70 ± 0.05 0.71 ± 0.02
C23:0 0.18 ± 0.02 y 0.28 ± 0.02 z 0.22 ± 0.01 y
ΣSFA 34.92 ± 1.25 33.88 ± 0.35 35.37 ± 1.69
C16:1 3.89 ± 0.22 3.63 ± 0.10 3.62 ± 0.01
C17:1 0.14 ± 0.02 0.17 ± 0.02 0.15 ± 0.01
C18:1c 0.20 ± 0.03 0.18 ± 0.02 0.19 ± 0.01
C18:1t 25.83 ± 0.17 z 23.80 ± 0.64 y 24.14 ± 0.47 y
C20:1 0.20 ± 0.02 0.21 ± 0.01 0.21 ± 0.03
C22:1(n-9) 0.26 ± 0.02 0.29 ± 0.04 0.26 ± 0.04
C24:1(n-9) 0.10 ± 0.02 0.10 ± 0.01 0.09 ± 0.02
ΣMUFA 30.60 ± 0.49 z 28.38 ± 1.13 y 28.64 ± 0.84 y
C18:2(n-6) 23.30 ± 1.08 y 25.21 ± 0.20 z 24.26 ± 0.90 zy
C18:3(n-6) 2.25 ± 0.11 2.53 ± 0.07 2.43 ± 0.09
C18:3(n-3) 1.63 ± 0.05 1.67 ± 0.06 1.65 ± 0.16
C20:2 0.85 ± 0.02 0.87 ± 0.07 0.83 ± 0.05
C20:3(n-6) 0.55 ± 0.03 0.52 ± 0.02 0.50 ± 0.03
C20:4(n-6) 0.47 ± 0.03 y 0.55 ± 0.03 z 0.54 ± 0.02 z
C20:5(n-3) 0.36 ± 0.03 0.38 ± 0.04 0.33 ± 0.04
C22:6(n-3) 7.06 ± 0.24 7.55 ± 0.33 7.08 ± 0.18
ΣPUFA 36.48 ± 1.37 y 39.28 ± 0.45 z 37.62 ± 1.17 zy
Σn-3 9.06 ± 0.24 9.60 ± 0.37 9.05 ± 0.24
Σn-6 26.57 ± 1.23 y 28.81 ± 0.38 z 27.73 ± 1.09 zy
n-6/n-3 0.34 ± 0.01 0.33 ± 0.02 0.32 ± 0.02

et al. (1997), Huang et al. (2015), and Baloi et al. (2017) digestive function were observed when GIFT were fed at
also reported that an increased daily feeding rate elevated different feeding rates. Harpaz et al. (2005) and Baloi et
plasma biochemical parameters in fish. These results sug- al. (2017) observed that fish fed low feeding rates had
gest that a higher feeding rate may cause some dysfunc- poor growth but higher digestive enzyme activities com-
tion in the metabolism of lipids and serious liver lesions in pared with those fed higher feeding rates, and this could
fish. be explained by the shortage of food. However, Sun et
Digestive enzyme activity is regarded as the depend- al. (2016) found that the digestive enzyme activities of
able indicator of fish nutritional state and fish feed fish fed to satiation were significantly higher than those
acceptance. Changes in feed intake and composition fed to 80% of satiation. Gisbert et al. (2011) reported
could influence the synthesis and secretion of digestive that feed deprivation resulted in a decrease in the specific
enzymes and further affect the rates of digestion and and total activities of intestinal brush border and pancre-
absorption of nutrients and the health of the gut atic enzymes and that activity fell more rapidly for
microenvironment of fish (Gisbert et al. 2011; Hoseinifar intestinal enzymes than for pancreatic enzymes, eventu-
et al. 2017). In this study, lipase, amylase, and protease ally causing a reduction in digestive function and mass
activities were not affected by feeding rate, which indi- loss. Gisbert et al. (2011) stated that the decrease in the
cated that no changes in digestive enzyme activities and intestinal enzyme activity might be attributed to a
 FEEDING RATES OF GENETICALLY IMPROVED FARMED NILE TILAPIA 7

reduction in the absorptive surface area and of the The n-6/n-3 ratio is a dependable indicator for compar-
intestinal mucosa, and the decrease in pancreatic enzyme ing the lipids nutritive values (Guler et al. 2008). A very
activity might be linked to an absence of digestive prod- high n-6/n-3 ratio and excessive amounts of n-6 polyunsat-
ucts that act directly on the pancreatic acinar cells to urated fatty acids may promote the pathogenesis of many
stimulate production and secretion of enzymes. diseases, such as inflammatory and autoimmune diseases,
Many studies have shown that there is a relationship cancer, and cardiovascular disease; however, an increased
between feeding rates and body composition of fish (Ng n-3 polyunsaturated fatty acid level (lower n-6/n-3 ratio) is
and Romano 2013; Karapanagiotidis 2016). In this study, more desirable in reducing the risk of these chronic dis-
moisture and crude protein in the muscle and crude pro- eases (Simopoulos 2008). In the present study, GIFT fed
tein in the whole body were not affected by feeding rates, at 3% BW/d had significantly lower C18:2(n-6) content,
but ash and moisture levels in the whole body were nega- while n-6/n-3 ratios (0.32–0.34), C22:6(n-3) (7.06–7.55%),
tively correlated with feeding rate. Generally, when C20:5(n-3) (0.33–0.38%), and C18:3(n-3) (1.63–1.67%)
farmed fish are fed at higher rates than that needed for exhibited no significant differences among all groups.
the maintenance requirement, the excess energy accumu- However, Rondán et al. (2004) found that the n-3/n-6
lates mainly in the form of lipid in the adipose tissues ratio and n-3 polyunsaturated fatty acids increased, while
(Lee et al. 2016), as shown in Common Carp (Shimeno et n-6 polyunsaturated fatty acids and C18:2(n-6) decreased,
al. 1997), Indian Major Carp (Abidi and Khan 2014; as food intake decreases, whether in perivisceral fat, white
Pradhan et al. 2018), Nile Tilapia (Huang et al. 2015), and muscle, or liver of Sharpsnout Seabream Diplodus pun-
GIFT (Liu et al. 2018). Similarly, we also observed that tazzo.
the crude lipid content in the whole body (5.78–7.25%)
and muscle (1.44–2.28%) increased as feeding rates Conclusion
increased, and these lipid values were lower than previous In conclusion, the present study has shown that growth,
results reported in tilapia Oreochromis sp. (Karapanagio- feed utilization, and body lipid content of GIFT were
tidis 2016). This may be due to the sustained swimming affected by feeding rates and that 5% BW/d is a suitable
exercise in a IPRS. Jobling et al. (1993) and Harimana et feeding level for GIFT fed a commercial feed twice daily
al. (2018) observed that the lipid level was reduced when in a IPRS.
fish were raised in lotic conditions compared with in lentic
conditions. A lower lipid level in muscle or whole body
could be explained by the aerobic metabolism using lipoly- ACKNOWLEDGMENTS
sis for energy production during sustained swimming exer- This work was supported by the Modern Agriculture
cise (Rasmussen et al. 2011). El-Saidy and Gaber (2005) Industry System Construction of Special Funds (CARS-
found that feeding rates had not affected the whole body 46) and the funds for Independent Innovation of Agricul-
energy and fat levels of Nile Tilapia, while increasing the tural Sciences of Jiangsu Province (CX[16]1004), and
feeding rate to 3% BW/d lead to the energy and lipid con- Three New Projects of Agricultural Aquaculture Program
tent reaching a highest value in flesh. However, Cho et al. of Jiangsu Province (D2016-18). There is no conflict of
(2007) reported that body composition seemed less interest declared in this article.
affected by feeding rates. These different results could be
due to differences in size of fish, sampling part, nutritional
history, and culture conditions (e.g., water temperature, REFERENCES
salinity) (Jobling et al. 1993; Rowland et al. 2005). Abidi, S. F., and M. A. Khan. 2014. Evaluation of feeding rate based on
In this study, the saturated fatty acid content was 33.88– growth, feed conversion, protein gain and carcass quality of fingerling
35.37% in the muscle of GIFT fed different feeding rates, Indian Major Carp, Catla catla (Hamilton). Aquaculture Research
and the main component in the saturated fatty acids was 45:439–447.
Ballantyne, J. S. 2001. Amino acid metabolism. Pages 77–107 in P.
palmitic acid, which coincides with earlier studies (Pradhan
Wright and P. Anderson, editors. Fish physiology. Academic Press,
et al. 2018; dos Santos et al. 2019). The GIFT fed 3% BW/d New York.
had lower linoleic acid content than those fed 4% and 5% Baloi, M. F., F. C. Sterzelecki, J. K. Sugai, G. Passini, C. V. A. Car-
BW/d. Similarly, Rondán et al. (2004) and Pradhan et al. valho, and V. R. Cerqueira. 2017. Growth performance, body compo-
(2018) also observed that linoleic acid content decreased sition and metabolic response to feeding rates in juvenile Brazilian
Sardine Sardinella brasiliensis. Aquaculture Nutrition 23:1458–1466.
along with feed intake; all these could be interpreted as this
Bhujel, R. C., D. C. Little, and A. Hossain. 2007. Reproductive perfor-
fatty acid being mobilized to obtain energy. The sum of mance and the growth of pre-stunted and normal Nile Tilapia (Ore-
polyunsaturated fatty acids (31.62–34.42%) increased, while ochromis niloticus) broodfish at varying feeding rates. Aquaculture
monounsaturated fatty acids (28.64–30.60%) decreased as 273:71–79.
feeding rates increased. Pradhan et al. (2018) observed simi- Brett, J. R., and T. D. D. Groves. 1979. Physiological energetics. Pages
279–352 in W. S. Hoar, D. J. Randall, and J. R. Brett, editors. Fish
lar results in Indian Major Carp.
8 WANG ET AL.

physiology: bioenergetics and growth, volume 8. Academic Press, protein and gross energy in the African Catfish Clarias gariepinus.
New York. Aquaculture 51:1–11.
Cho, C. Y., and D. P. Bureau. 2001. A review of diet formulation strate- Hoseinifar, S. H., M. Dadar, and E. Ringø. 2017. Modulation of nutri-
gies and feeding systems to reduce excretory and feed wastes in aqua- ent digestibility and digestive enzyme activities in aquatic animals: the
culture. Aquaculture Research 32:349–360. functional feed additives scenario. Aquaculture Research 48:3987–
Cho, S. H., S. M. Lee, B. H. Park, S. C. Ji, C. Y. Choi, J. H. Lee, Y. C. 4000.
Kim, J. H. Lee, and S. Y. Oh. 2007. Effect of daily feeding ratio on Huang, Q., K. Huang, Y. Ma, X. Qin, Y. Wen, L. Sun, and L. Tang.
growth and body composition of subadult Olive Flounder, Par- 2015. Feeding frequency and rate effects on growth and physiology of
alichthys olivaceus, fed an extruded diet during the summer season. juvenile genetically improved farmed Nile Tilapia. North American
Journal of the World Aquaculture Society 38:68–73. Journal of Aquaculture 77:503–512.
Cho, S. H., S. M. Lee, B. H. Park, and S. M. Lee. 2006. Effect of feed- Jobling, M., B. M. Baardvik, J. S. Christiansen, and E. H. Jørgensen.
ing ratio on growth and body composition of juvenile Olive Flounder 1993. The effects of prolonged exercise training on growth perfor-
Paralichthys olivaceus fed extruded pellets during the summer season. mance and production parameters in fish. Aquaculture International
Aquaculture 251:78–84. 1:95–111.
Clark, J. H., W. O. Watanabe, D. H. Ernst, R. I. Wicklund, and B. L. Karapanagiotidis, I. T. 2016. Nutrient profiles of tilapia. Pages 261–305
Olla. 1990. Effect of feeding rate on growth and feed conversion of in P. W. Perschbacher and R. R. Stickney, editors. Tilapia in inten-
Florida red tilapia reared in floating marine cages. Journal of the sive co-culture. Wiley, Hoboken, New Jersey.
World Aquaculture Society 21:16–24. Lee, S., L. Y. Haller, N. A. Fangue, J. G. Fadel, and S. S. O. Hung.
Craig, S. 2009. Understanding fish nutrition, feeds, and feeding. Virginia 2016. Effects of feeding rate on growth performance and nutrient par-
Polytechnic Institute and State University, Publication 420-256, titioning of young of the year White Sturgeon. Aquaculture Nutrition
Blacksburg. 22:400–409.
De Silva, S. S., M. R. Gunasekera, and C. Keembiyahetty. 1986. Opti- Lewis, D. E., L. Y. N. D. S. E. Y. Shellard, D. G. Koeslag, D. E. Boer,
mum ration and feeding frequency in Oreochromis niloticus young. H. D. McCarthy, P. E. McKibbin, and G. Williams. 1993. Intense
Pages 559–564 in J. L. Maclean, L. B. Dizon, and L. V. Hosillos, edi- exercise and food restriction cause similar hypothalamic neuropeptide
tors. The first Asian fisheries forum. Asian Fisheries Society, Manila. Y increases in rats. American Journal of Physiology Endocrinology
Dong, J., Y. Y. Zhao, Y. H. Yu, N. Sun, Y. D. Li, H. Wei, Z. Q. and Metabolism 264:E279–E284.
Yang, X. D. Li, and L. Li 2018. Effect of stocking density on Li, X. P., J. R. Li, Y. B. Wang, L. L. Fu, Y. Y. Fu, B. Q. Li, and B. H.
growth performance, digestive enzyme activities, and nonspecific Jiao. 2011. Aquaculture industry in China: current state, challenges,
immune parameters of Palaemonetes sinensis. Fish and Shellfish and outlook. Reviews in Fisheries Science 19:187–200.
Immunology 73:37–41. Lim, C., C. D. Webster, and C. S. Lee. 2015. Feeding practices and fish
dos Santos, S. K. A., M. Schorer, G. D. S. Moura, E. A. T. Lanna, and health. Pages 333–346 in C. Lee, C. Lim, D. M. Gatlin, and C. D.
M. M. Pedreira. 2019. Evaluation of growth and fatty acid profile of Webster, editors. Dietary nutrients, additives, and fish health. Wiley,
Nile Tilapia (Oreochromis niloticus) fed with Schizochytrium sp. Hoboken, New Jersey.
Aquaculture Research 50:1068–1074. Liu, W., H. Wen, and Z. Luo. 2018. Effect of dietary protein levels and
El-Saidy, D. M. S. D., and M. M. A. Gaber. 2005. Effect of dietary pro- feeding rates on the growth and health status of juvenile genetically
tein levels and feeding rates on growth performance, production traits improved farmed tilapia (Oreochromis niloticus). Aquaculture Interna-
and body composition of Nile Tilapia, Oreochromis niloticus (L.) cul- tional 26:153–167.
tured in concrete tanks. Aquaculture Research 36:163–171. Melzer, K., B. Kayser, W. H. Saris, and C. Pichard. 2005. Effects of
El-Sayed, A. F. M. 2002. Effects of stocking density and feeding levels physical activity on food intake. Clinical Nutrition 24:885–895.
on growth and feed efficiency of Nile Tilapia (Oreochromis niloticus Naylor, R. L., R. W. Hardy, D. P. Bureau, A. Chiu, M. Elliott, A. P.
L.) fry. Aquaculture Research 33:621–626. Farrell, I. Forster, D. M. Gatlin, F. J. Goldburg, K. Hua, and P. D.
Gisbert, E., I. Fernández, and C. A. Alvarez-González. 2011. Prolonged Nichols. 2009. Feeding aquaculture in an era of finite resources. Pro-
feed deprivation does not permanently compromise digestive function ceedings of the National Academy of Sciences of the USA
in migrating European glass eels Anguilla anguilla. Journal of Fish 106:15103–15110.
Biology 78:580–592. Ng, W. K., and N. Romano. 2013. A review of the nutrition and feeding
Guler, G. O., B. Kiztanir, A. Aktumsek, O. B. Citil, and H. Ozparlak. management of farmed tilapia throughout the culture cycle. Reviews
2008. Determination of the seasonal changes on total fatty acid com- in Aquaculture 5:220–254.
position and ω3/ω6 ratios of carp (Cyprinus carpio L.) muscle lipids in Öner, M., G. Atli, and M. Canli. 2008. Changes in serum biochemical
Beysehir Lake (Turkey). Food Chemistry 108:689–694. parameters of freshwater fish Oreochromis niloticus following pro-
Hardy, R. W. 1998. Feeding salmon and trout. Pages 175–197 in T. longed metal (Ag, Cd, Cr, Cu, Zn) exposures. Environmental Toxi-
Lovell, editor. Nutrition and feeding of fish. Kluwer Academic Pub- cology and Chemistry 27:360–366.
lishers, Boston. Pradhan, C., S. S. Giri, S. N. Mohanty, and R. Narasimmalu. 2018.
Harimana, Y., X. Tang, G. Le, X. Xing, K. Zhang, Y. Sun, and M. A. Effect of different feeding levels of plant-ingredient-based feed on fillet
Tuyishimire. 2018. Quality parameters of Black Carp (Mylopharyn- fatty acid profile, carcass trait, and sensory characteristics of Indian
godon piceus) raised in lotic and lentic freshwater systems. LWT- Major Carps in earthen pond polyculture. Journal of the World
Food Science and Technology 90:45–52. Aquaculture Society 50:374–389.
Harpaz, S., Y. Hakim, A. Barki, I. Karplus, T. Slosman, and O. T. Puvanendran, V., D. L. Boyce, and J. A. Brown 2003. Food ration
Eroldogan. 2005. Effects of different feeding levels during day and/or requirements of 0 + Yellowtail Flounder Limanda ferruginea (Storer)
night on growth and brush-border enzyme activity in juvenile Lates juveniles. Aquaculture 220:459–475.
calcarifer reared in freshwater re-circulating tanks. Aquaculture Rasmussen, R. S., M. T. Heinrich, G. Hyldig, C. Jacobsen, and A.
248:325–335. Jokumsen. 2011. Moderate exercise of Rainbow Trout induces only
Henken, A. M., D. W. Kleingeld, and P. A. T. Tijssen. 1985. The effect minor differences in fatty acid profile, texture, white muscle fibres and
of feeding level on apparent digestibility of dietary dry matter, crude proximate chemical composition of fillets. Aquaculture 314:159–164.
 FEEDING RATES OF GENETICALLY IMPROVED FARMED NILE TILAPIA 9

Ribeiro, F. F., S. Forsythe, and J. G. Qin. 2015. Dynamics of intraco- and nutritional utilization of the diet. Aquaculture Nutrition 12:403–
hort cannibalism and size heterogeneity in juvenile Barramundi at dif- 409.
ferent stocking densities and feeding frequencies. Aquaculture 444: Wang, Q. D., L. Cheng, J. S. Liu, Z. J. Li, S. Q. Xie, and S. S. De Silva.
55–61. 2015. Freshwater aquaculture in PR China: trends and prospects.
Rondán, M., M. D. Hernández, M. Á. Egea, B. Garcia, F. M. Rueda, Reviews in Aquaculture 7:283–302.
and F. J. Martinez. 2004. Effect of feeding rate on fatty acid compo- Wang, Y., P. Xu, Z. Nie, Q. Li, N. Shao, and G. Xu. 2019. Growth,
sition of Sharpsnout Seabream. Aquaculture Nutrition 10:301–307. digestive enzymes activities, serum biochemical parameters and
Rowland, S. J., G. L. Allan, C. Mifsud, M. Nixon, P. Boyd, and D. antioxidant status of juvenile genetically improved farmed tilapia
Glendenning. 2005. Development of a feeding strategy for Silver (Oreochromis niloticus) reared at different stocking densities in in-
Perch, based on restricted rations. Aquaculture Research 36:1429– pond raceway recirculating culture system. Aquaculture Research
1441. 50:1338–1347.
Satheeshkumar, P., G. Ananthan, D. Senthilkumar, A. B. Khan, and K. Wunderink, Y. S., G. Martínez-Rodríguez, M. Yúfera, I. M. Montero,
Jeevanantham. 2012. Comparative investigation on haematological and G. Flik, J. M. Mancera, and P. H. Klaren. 2012. Food deprivation
biochemical studies on wild marine teleost fishes from Vellar estuary, induces chronic stress and affects thyroid hormone metabolism in
southeast coast of India. Comparative Clinical Pathology 21:275–281. Senegalese Sole (Solea senegalensis) post-larvae. Comparative Bio-
Shimeno, S., T. Shikata, H. Hosokawa, T. Masumoto, and D. Kheyyali. chemistry and Physiology Part A: Molecular and Integrative Physiol-
1997. Metabolic response to feeding rate in Common Carp, Cyprinus ogy 162:317–322.
carpio. Aquaculture 151:371–377. Xu, P., and J. C. Ming. 2018. Status and trends of the tilapia farming
Simopoulos, A. P. 2008. The importance of the omega-6/omega-3 fatty industry development. Pages 404–420 in J. F. Gui, Q. S. Tang, Z. J.
acid ratio in cardiovascular disease and other chronic diseases. Exper- Li, J. S. Liu, and S. S. De Silva, editors. Aquaculture in China: suc-
imental Biology and Medicine 233:674–688. cess stories and modern trends. Wiley, Hoboken, New Jersey.
Sun, G., Y. Liu, D. Qiu, M. Yi, X. Li, and Y. Li. 2016. Effects of feed- Yuan, Y. C., H. J. Yang, S. Y. Gong, Z. Luo, H. W. Yuan, and X. K.
ing rate and frequency on growth performance, digestion and nutri- Chen 2010. Effects of feeding levels on growth performance, feed uti-
ents balances of Atlantic Salmon (Salmo salar) in recirculating lization, body composition and apparent digestibility coefficients of
aquaculture systems. Aquaculture Research 47:176–188. nutrients for juvenile Chinese Sucker, Myxocyprinus asiaticus. Aqua-
Tandon, R. S., and S. Chandra. 1976. Serum alkaline phosphatase levels culture Research 41:1030–1042.
of some fresh water teleosts. Zeitschrift für Tierphysiologie Tier- Zikic, R. V., S. Stajn, Z. Pavlovic, B. I. Ognjanovic, and Z. S. Saicic.
ernährung und Futtermittelkunde 37:330–333. 2001. Activities of superoxide dismutase and catalase in erytrocyte
Velázquez, M., S. Zamora, and F. J. Martínez. 2006. Effect of different and plasma transaminases of Goldfish (Carassius auratus gibelio
feeding strategies on Gilthead Seabream demand-feeding behavior Bloch.) exposed to cadmium. Physiological Research 50:105–111.