Effects of α-Tocopherol Supplemented Corn Gluten Meal
Based Diet on Growth, Nutrient Digestibility and Antioxidant Activity of Catla catla Fingerlings
Syed Makhdoom
Hussain1†*, Muhammad Arshad1†, Shafaqat
Ali2,3, Muhammad Moazam Jalees4, Farooq Ahmad5, Fatima
Bashir1, Aqsa Sharif1 and
Zeeshan Yousaf1
1Fish Nutrition Lab,
Department of Zoology, Government College University, Faisalabad, Pakistan
2Department of Environmental Sciences & Engineering,
Government College University, Faisalabad, Pakistan
3Department of Biological Sciences and Technology,
China Medical University, Taichung 40402, Taiwan
4Department of Zoology, The
Islamia University of Bahawalpur, Pakistan
5Department of
Microbiology, Cholistan University of Veterinary and Animal Sciences Bahawalpur
*For Correspondence: drmakhdoom90@gmail.com
Received 20 January 2021; Accepted 12 June 2021; Published 18 September 2021
Abstract
A feeding trial
of 70 days was conducted to assess the effects of α-tocopherol on
growth performance, antioxidant activity and nutrient digestibility of C.
catla fingerlings (7.05 ± 0.02 g body weight and
6.05 ± 1.07 cm body length) were fed on corn gluten meal-based diets. The α-tocopherol
was supplemented at various concentrations viz., 0, 100, 200, 300, 400,
500 and 600 mg/kg of body weight. Triplicate tanks were used for feeding trial
and each tank housed 15 fingerlings. During trial, fingerlings were fed at 5%
of their body weight. Completely randomized design (CRD) was followed to
allocate test diets to the fish groups. Our results showed that 300 mg/kg of α-tocopherol was the optimal
level that had a significant role in improving growth parameters such as weight
gain (230%) and feed conversion ratio (1.80). Nutrient digestibility parameters
i.e., crude protein (75.79%),
crude fat (76.54%) and gross energy (61.03%) were best observed at the same
level compared with the control and other test diets. Furthermore,
antioxidant status of the fish was improved with the increase in concentration
of α-tocopherol,
so, only 5.46% oxidation was observed at 600 mg/kg. It was concluded from present
project that corn gluten meal-based diet supplemented with 300 mg/kg level of
α-tocopherol proved to be effective in developing low cost, nutritionally
balanced and environment friendly feed for C.
catla. © 2021 Friends Science Publishers
Keywords: α-tocopherol; Antioxidant activity; Aquaculture; Corn gluten meal; Growth
Introduction
Nowadays, fish
farming is becoming the top growing industry for producing good quality food.
However, we are facing challenges in improving aquaculture sector like high
demand of fish products and limited supply of fish feed ingredients especially
fish meal (Zarantoniello
et al. 2020). Along with
nutritional values, this sector also provides employment for millions of the
people especially belonging to poor and rural areas (Salim 2006). The main
target for most of the fish nutritionists is to prepare good quality feed for
fish at minimum cost. It is estimated that cost of feed is 50 to 60% of the
whole expenditure of aquaculture (Shahzad et al.
2018). Requirement of fish meal due
to increase in its cost and shortage made it essential to look for the other
sources of protein for fish feed (Pham et al. 2008). To replace fish meal, appropriate plant feed
ingredients such as grains and other plant by-products are being used as the
most favorable source of protein for feed formulation (Manuel et al. 2019). Corn gluten meal is
produced during processing of corn starch, contains approximately 60% protein
and has potential to replace fish meal in diet because of its essential amino
acid profile (Men et al.
2014).
The substances which reduce the reactive oxygen species
are called as antioxidants (Majid et al.
2015). Under normal physiological conditions, the body of animals has capacity
to produce suitable quantity of antioxidants that are able to prevent the
process of oxidation. When there is higher rate of lipid per-oxidation,
reactive oxygen specie is produced constantly. When free radicals exceed the
limit of antioxidant potential of the body, stress is produced by oxidative
response in body (Rahman et al.
2014). Vitamins are the organic compounds required by organisms in minute
amount for proper growth and metabolism (Amlashi et al. 2012). Since most of
fishes cannot synthesize vitamins at all or can only produce them in small
quantities for normal growth and metabolism, they must be supplemented in the
diet (Falahatkar et al. 2011).
Vitamin E is required to maintain immunity, save cell membrane from peroxide
damage and improves the tissue feature (Salinthone et al. 2013). Vitamin E is composed of
four tocopherols and four tocotrienols having two rings and amongst them the
most common and biologically dynamic molecule is α-tocopherol (Yang
et al. 2012).
Fish fed with vitamin E showed significantly higher growth
rate and feed efficiency (Kashani et al. 2010). Vitamin E
requirement have been confirmed in many fish species. It was basically
considered as a dietary component in nutrition of animals, which is very
important for fish reproduction. In aquaculture, it is added in feed to enhance
growth, and survival of fish against disease and increase stress resilience (Vismara et al.
2003). C. catla is an important carp specie; being cultured extensively
for its fast growth rate, nutritional quality, good taste, acclimatization to
laboratory conditions and high commercial value (Wahab et al. 2002). The present research
work was conducted to evaluate the effect of α-tocopherol
on growth performance, nutrient digestibility and antioxidant activity of C. catla fingerlings.
Materials and
Methods
Present project was carried out in Fish Nutrition
Laboratory, Department of Zoology, Government College University, Faisalabad,
Pakistan. C. catla fingerlings of the
equal body weight (7.05 ± 0.02 g) and same body length (6.05 ± 1.07 cm) were
purchased from Government Fish Hatchery, Satiana road, Faisalabad.
Fish
acclimatization and culture conditions
Fingerlings were kept in V-shaped tanks having 70 L
volumetric capacity (specifically made for feces collection) and were acclimatized
to experimental conditions for 15 days (Rowland 1991). Fingerlings were fed
once with basal diet for two weeks (Allan and Rowland 1992). Before experiment,
C. catla fingerlings were bathed in
0.5% NaCl solution, so as to free fish from fungal infection and
ecto-parasites. On daily basis, temperature was measured with thermometer, pH
with pH meter (Jenway 3510) and dissolved oxygen with DO meter (Jenway 970). By
capillary method, aeration (24 h) was supplied to all tanks.
Feed
ingredients and experimental diets preparation
Feed ingredients such as corn
gluten meal, fish meal and soybean meal were grinded by passing through
Feeding protocol
Feeding trial of 70 days was conducted and fingerlings
were fed @ of 5% of their wet body weight, two times a day. Remaining diet was
drained out from each tank after the feeding period of two hours on daily
basis; by opening the valves of tanks. To remove the remaining diet particles,
tanks were washed entirely and filled with water again. Feces were collected
through fecal collecting tube of each tank through valve-I and II. To minimize
the leakage of nutrients, fecal strings were collected carefully to avoid fecal
dissolving. The collected strings were first dried in oven and then grinded for
further chemical analysis.
Growth parameters
To study
growth rate, fish in each tank were bulk weighed at the start and at the end of
whole experimental period. Weight gain (WG %), FCR and specific growth rate
(SGR) of fingerlings were checked by using the following standard formulae (National
Research Council 1993) :
Nutrient digestibility
Evaluation of
apparent nutrient digestibility coefficients (ADC%) of experimental diets were
measured by using the standard formula (National Research Council 1993):
Antioxidant activity
To determine antioxidant status of C. catla
fingerlings, percent inhibition of oxidation was calculated following Hussain et
al. (2011) with minor modifications. 1 g dried and ground samples of fish
body from each group were mixed with 10 mL of n-hexane in separate test tubes.
Following this, test tubes having hexane fraction were heated gently in water
bath for about 10 min. To prepare 10 mL solution of 0.2 M, phosphate buffer was added in each test tube. After gently
shaking, 200 μL from each test
tube was picked and transferred to a new one so that 200 μL of 30% aqueous ammonium thiocyanate solution and 200 μL of 35% ferrous chloride solution
can be mixed in it. At the end, absorbance of solution by adding 10 mL of 95%
ethanol in each test tube was measured using spectrophotometer at 500 nm. The
percentage inhibition of oxidation was evaluated using the formula:
% inhibition = [(A0 - As) /A0] × 100
In formula, A
(0) and A(s) are the sample’s absorbance from 0 to 5 min, respectively.
Statistical
analysis
One way analysis of variance (ANOVA) was applied on the data of growth
performance, nutrient digestibility and anti-oxidant status. The
differences between means were compared by applying Tukey’s Test (Snedecor and
Cochran 1991). For statistical analysis, the Co-Stat computer software (version
6.303, Monterey, CA, PMB 320, 93940 USA) was used.
Results
Growth parameters
The growth performance of C. catla fingerlings fed on corn gluten meal-based diets
supplemented with different levels of α-tocopherol as 0, 100, 200, 300,
400, 500 and 600 mg/kg diet is shown in Table 1, 2. In terms of
weight gain %, diet IV with
α-tocopherol at 300 mg/kg of diet showed maximum values. In all
treatments, initial weight of fingerlings was almost same, but weight of
fingerlings measured after experiment was significantly different than others.
The maximum weight gain (16.31 g), weight gain % (230%) was noted in fish fed
at 300 mg/kg of α-tocopherol supplementation, followed by fish (15.51 g;
219%) fed at 200 mg/kg of α-tocopherol. These values were found to be
significantly (P < 0.05)
different when compared with control diet (12.92 g; 183.35%) and other test
diets. Beyond 300 mg/kg, further increase in α-tocopherol level did not
result in improved results. Its maximum concentration improved weight gain only up to 171%. Lowest value of FCR (1.80) was noticed
in fish fed diet having 300 mg/kg α-tocopherol which rendered most of the
feed portion to be converted in body flesh. Alternatively, highest FCR value
(1.99) was obtained by using control diet having no supplementation of vitamin
E. Supplementation of α-tocopherol in corn gluten meal-based diet at the
level of 300 mg/kg had a vital role in promoting growth of fish.
Nutrient digestibility
The analyzed composition of nutrients (crude protein,
crude fat and gross energy) in diet and feces of C. catla fingerlings fed on graded concentrations
of α-tocopherol supplemented corn gluten meal-based diet is shown in Table
3 and 4, respectively. It was
observed that α-tocopherol at 300 mg/kg supplementation in corn gluten
meal-based diet played a vital role in improving apparent digestibility
coefficient (ADC%). Results showed that increasing trend was observed in
nutrients digestibility up to level IV having 300 mg/kg of
α-tocopherol in diet where it reached its maximum, while further increase
in α-tocopherol supplementation resulted in reduced digestibility of
nutrients. In current study, it was observed that at 300 mg/kg level,
the digestibility value of crude fat (76.54%), crude
protein (75.79%) and gross energy (61.03%) were found to be highest and significantly (P < 0.05) different than all other
test diets (Table 5). However, lowest digestibility value of nutrients, crude
fat (64.68%), crude protein (55.36%) and gross energy (40.62%) was noted at control diet (0 mg/kg α-tocopherol).
These results revealed that supplementation of α-tocopherol at the level
of 300 mg/kg of diet is the best for maximum digestibility of nutrients in body
of C. catla fingerlings and resulted
in reduced discharge of nutrients in environment.
Antioxidant activity
The α-tocopherol’s antioxidant activity at various
levels (0, 100, 200, 300, 400, 500 and 600 mg/kg) was checked as shown in Table
6. Decreasing trend of percentage of oxidation was observed with increasing
level of α-tocopherol in all fish groups. It means that antioxidant
activity of α-tocopherol was best described at its highest level.
Experimental diet VII with 600 mg/kg of α-tocopherol was observed to be
the best (P < 0.05) of all other
test diets because the percentage of oxidation was lowest (5.46%) when compared
with other test diets. Second good results (18.22%) were found at 500 mg/kg
level of α-tocopherol. On the other hand, 100% oxidation was observed in
control group that fed on diet supplemented with 0 mg/kg of α-tocopherol.
Discussion
The present study showed that α-tocopherol is an important
feed additive to enhance antioxidant activity, improve growth and nutrient
digestibility. The higher weight gain, weight
gain values were observed in fingerlings fed on 300 mg/kg of alpha tocopherol
supplemented corn gluten meal-based diet. Results resembling to our study were reported by Muchlisin et al.
(2016) who described that 150 mg/kg of α-tocopherol is an
optimum dosage for better growth of keureling (Tor tambra). Galaz et al. (2010) found that increasing the dose of dietary α-tocopherol (up
to 500 mg/kg) enhanced growth in Vibrio anguilarum challenged fish
(Nekoubin et al. 2012). Gao et al. (2012) demonstrated that vitamin
C supplementation
in red sea bream at 500 mg/kg level could improve growth parameters and can
reduce oxidative stress. However, in contradiction to current results, Gao et al. (2014) reported that the
increasing dietary vitamin E level from 100 to 200 mg/kg decreased growth
performance in juvenile Japanese flounder, Paralichthys
olivaceus.
Table 1: Ingredients composition (%) of test diets
Ingredients |
Test Diet-I (Control) |
Test Diet-II |
Test Diet-III |
Test Diet-IV |
Test Diet-V |
Test Diet-VI |
Test Diet-VII |
α-tocopherol*
(mg/kg) |
0 |
100 |
200 |
300 |
400 |
500 |
600 |
Corn gluten
meal |
55 |
55 |
55 |
55 |
55 |
55 |
55 |
Fish meal |
16 |
16 |
16 |
16 |
16 |
16 |
16 |
Wheat flour |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
Soybean
meal |
8 |
8 |
8 |
8 |
8 |
8 |
8 |
Fish oil |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
Vitamin
Premix |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Mineral
mixture |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Ascorbic
acid |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Chromic
oxide |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Total |
100 |
100 |
100 |
100 |
100 |
100 |
100 |
*α-tocopherol was supplemented at the cost of wheat flour
Table 2: Growth
performance of C. catla
fingerlings fed α-tocopherol supplemented
corn gluten meal-based diets
Growth parameters |
Test
Diet-I (Control diet) |
Test
Diet-II |
Test
Diet-III |
Test
Diet-IV |
Test
Diet-V |
Test
Diet-VI |
Test
Diet-VII |
α-tocopherol Levels (mg/kg) |
|
||||||
0 |
100 |
200 |
300 |
400 |
500 |
600 |
|
IW (g) |
7.07 ± 0.03 a |
7.06 ± 0.02a |
7.09 ± 0.02 a |
7.09 ± 0.03a |
7.05 ± 0.05a |
7.06 ± 0.02 a |
|
FW (g) |
19.97 ± 0.02e |
21.41 ± 0.03c |
22.58 ± 0.06b |
23.40 ± 0.04a |
21.46 ± 0.06c |
20.94 ± 0.05d |
19.19 ± 0.03f |
WG (g) |
12.92 ± 0.03e |
14.35 ± 0.01c |
15.51 ± 0.05c |
16.31 ± 0.03b |
14.36 ± 0.04a |
13.89 ± 0.05c |
12.12 ± 0.02b |
WG (%) |
183.35 ± 0.78c |
203.02 ± 0.64b |
219.63 ± 0.86a |
230.00 ± 0.58a |
202.49 ± 0.50b |
196.89 ± 0.09b |
171.64 ± 0.42d |
WG
(fish/day) g |
0.18 ± 0.00d |
0.20 ± 0.00d |
0.22 ± 0.00cd |
0.23 ± 0.00b |
0.21 ± 0.00a |
0.20 ± 0.00bc |
0.17 ± 0.00d |
FI |
0.37 ± 0.02bc |
0.39 ± 0.03cd |
0.41 ± 0.02ab |
0.42 ± 0.01a |
0.38 ± 00.2a |
0.39 ± 0.01ab |
0.34 ± 0.01c |
FCR |
1.99 ± 0.08ab |
1.89 ± 0.12abc |
1.85 ± 0.10ab |
1.80 ± 0.05ab |
1.84 ± 0.08ab |
1.97 ± 0.05a |
1.98 ± 0.04ab |
Means within rows having different superscripts are significantly
different at P < 0.05
Data are means of three replicates
IW= Initial Weight, FW= Final Weight, WG= Weight gain,
FCR= Feed Conversion Ratio, FI= Feed Intake
Table 3: Analyzed
compositions (%) of CP, EE and GE in feed of C. catla fingerlings fed on corn gluten
meal-based diet with α-tocopherol supplemented test diets
Experimental diets |
α-tocopherol Levels (mgkg-1) |
CP (%) |
EE (%) |
GE (kcalg-1) |
Test Diet-I (Control) |
0 |
30.87 ± 0.02a |
7.70 ± 0.14bc |
4.12 ± 0.01a |
Test Diet-II |
100 |
30.87 ± 0.02a |
7.73 ± 0.08b |
4.26 ± 0.04a |
Test Diet-III |
200 |
30.89 ± 0.02a |
7.81 ± 0.06a |
4.30 ± 0.03a |
Test Diet-IV |
300 |
30.88 ± 0.03a |
7.90 ± 0.06b |
4.23 ± 0.02a |
Test Diet-V |
400 |
30.86 ± 0.02a |
7.82 ± 0.06bc |
4.26 ± 0.03a |
Test Diet-VI |
500 |
30.88 ± 0.02a |
7.81 ± 0.07bc |
4.23 ± 0.02a |
Test Diet-VII |
600 |
30.87 ± 0.02a |
7.80 ± 0.06c |
4.26 ± 0.03ab |
CP= crude protein, EE= Ether Extract, GE= Gross Energy
Means within columns having different superscripts are
significantly different at P < 0.05
Data are means of three replicates
The accessibility of
nutrients to the fish is basically related with its digestibility values. In current
study, it was observed that at 300 mg/kg level, the digestibility values of
crude fat (76.54%), crude protein (75.79%) and gross energy (61.03%) were found to
be highest and significantly different from all other test diets. Muchlisin et al.
(2016) reported that dietary vitamin E significantly affected protein retention
in the carcass and protein digestibility of fish. Chae et al. (2006) concluded that supplementation of 100 and 200 mg/kg
of α-tocopherol increased the nutrient digestibility in chicks. Data
about nutrient digestibility in fish fed α-tocopherol or vitamin E is rare.
Vitamin E possesses effective antioxidant activity that
protects fish tissues from oxidative damage (Rainis et al. 2007). Ciocoiu et al. (2007) deduced that α-tocopherol performed major
antioxidant activity in per oxidation of lipids in mammalian plasma membrane.
Significant role of α-tocopherol in tissues appeared to be protecting the
membrane polyunsaturated fatty acids from oxidation of free radicals. Tocher et al. (2002) concluded that decreased level of α-tocopherol in tissues of
fish reduced the activity of liver antioxidant enzymes and increased the
activity of lipid peroxide enzymes in juvenile turbot (Scophthalmus
maximus L.), halibut (Hippoglossus hippoglossus L.) and red sea
bream (Sparus aurata L.). From these results, it can be concluded that vitamin E played
an important role against oxygen free radicals to protect biological membranes.
Conclusion
Results of present research work suggested that supplementation
of α-tocopherol in corn gluten meal-based diet improved the growth
parameters, nutrient digestibility and antioxidant activity of C. catla. Maximum values of growth
performance, nutrient digestibility and antioxidant activity analysis in C. catla were observed at 300 mg/kg of
α-tocopherol level.
Acknowledgements
Table 4: Analyzed
compositions (%) of CP, EE and GE in feces of C. catla fingerlings fed on corn gluten
meal-based diet with α-tocopherol supplemented test diets
Experimental diets |
α-tocopherol Levels (mgkg-1) |
CP (%) |
EE (%) |
GE (kcalg-1) |
Test Diet-I (Control) |
0 |
14.28 ± 0.10a |
2.92 ± 0.054a |
2.69 ± 0.16a |
Test Diet-II |
100 |
9.18 ± 0.03e |
2.70 ± 0.02b |
2.26 ± 0.08ab |
Test Diet-III |
200 |
9.27 ± 0.02e |
2.20 ± 0.03e |
2.07 ± 0.64ab |
Test Diet-IV |
300 |
8.41 ± 0.05f |
1.97 ± 0.05f |
1.82 ± 0.10b |
Test Diet-V |
400 |
11.48 ± 0.34d |
2.15 ± 0.02e |
1.96 ± 0.01b |
Test Diet-VI |
500 |
12.87 ± 0.09c |
2.46 ± 0.03d |
2.27 ± 0.06ab |
Test Diet-VII |
600 |
13.75 ± 0.10b |
2.57 ± 0.02c |
2.46 ± 0.13ab |
CP= crude protein, EE= Ether Extract, GE= Gross Energy
Means within columns having different superscripts are
significantly different at P < 0.05
Data are means of three replicates
Table 5: Apparent
digestibility coefficient (%) of CP, EE and GE of C. catla fingerlings fed on corn gluten
meal-based diet with α-tocopherol supplemented test diets
Experimental diets |
α-tocopherol Levels (mgkg-1) |
CP (%) |
EE (%) |
GE (%) |
Test Diet-I (Control) |
0 |
55.36 ± 0.19g |
64.68 ± 0.33d |
40.62 ± 0.78b |
Test Diet-II |
100 |
71.13 ± 0.24c |
66.20 ± 0.86d |
51.21 ± 0.23ab |
Test Diet-III |
200 |
72.82 ± 0.19b |
72.82 ± 0.57b |
54.21 ± 0.47ab |
Test Diet-IV |
300 |
75.79 ± 0.72a |
76.54 ± 0.64a |
61.03 ± 0.39a |
Test Diet-V |
400 |
66.82 ± 0.89d |
74.29 ± 0.35ab |
57.58 ± 0.60a |
Test Diet-VI |
500 |
62.84 ± 0.71e |
70.28 ± 0.39c |
50.97 ± 0.73ab |
Test Diet-VII |
600 |
59.89 ± 0.56f |
65.92 ± 0.80d |
46.45 ± 0.85ab |
CP= crude protein, EE= Ether Extract, GE= Gross Energy
Means within columns having different superscripts are
significantly different at P < 0.05
Data are means of three replicates
Table 6: Antioxidant
activity of α-tocopherol of C. catla fingerlings fed on corn gluten meal-based diet
with α-tocopherol supplemented test diets
Experimental diets |
α-tocopherol Levels (mgkg-1) |
Absorbance |
Oxidation (%) |
Test Diet-I (Control) |
0 |
0.0283 ± 0.00012 |
100.00 ± 0.00 |
Test Diet-II |
100 |
0.0274 ± 0.00010 |
97.81 ± 0.35 |
Test Diet-III |
200 |
0.0261 ± 0.00019 |
93.34 ± 0.74 |
Test Diet-IV |
300 |
0.0244 ± 0.00013 |
84.28 ± 0.82 |
Test Diet-V |
400 |
0.0078 ± 0.00022 |
33.25 ± 0.44 |
Test Diet-VI |
500 |
0.0054 ± 0.00015 |
18.22 ± 0.34 |
Test Diet-VII |
600 |
0.0013 ± 0.00011 |
5.46 ± 0.24 |
Means within columns having different superscripts are
significantly different at P < 0.05
Data are means of three replicates
The authors would like to express their
acknowledgements toHEC Pakistan for providing funds for Project #
5649/Punjab/NRPU/R&D/HEC/2016 and Fish Nutrition Laboratory, Department of
Zoology, Government College University, Faisalabad for providing all type of
facilities required for this research work.
Author Contributions
Syed Makhdoom Hussain Conceived and
designed the project and provided facilities for research work; Muhammad Arshad
Conducted trial, collected data and wrote this manuscript; Shafaqat Ali Helped
in writing, revising and improvement of manuscript; Muhammad Moazam Jalees
Helped in writing the manuscript; Farooq Ahmad helped in writing, revising and
improvement of manuscript; Fatima Bashir Helped in compiling results; Aqsa
Sharif Helped in nutrients analysis; Zeeshan Yousaf Helped in conducting
feeding trial and formulating fish feed.
Conflict of Interest
All authors declare no conflict of
interest
Data Availability
Data presented in this study will be
available on a fair request to the corresponding autho
Ethical Approval
Not applicable in this paper.
Funding Source
HEC NRPU Project No. 5649/Punjab/NRPU/R&D/HEC/2016
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