DOI: 10.1111/are.

13735

ORIGINAL ARTICLE

Replacement of de-oiled rice bran by soaked and fermented

sweet potato leaf meal: Effect on growth performance, body
composition and expression of insulin-like growth factor 1 in
Labeo rohita (Hamilton), fingerlings

Shweta Meshram | Ashutosh D Deo | Sarvendra Kumar | Md Aklakur | Narottam

P Sahu

Fish Nutrition, Biochemistry and Physiology

Division, ICAR-Central Institute of Fisheries
Education, Versova, Mumbai, India
Correspondence
Ashutosh D. Deo, Fish Nutrition,
Biochemistry and Physiology Division, ICAR-
Central Institute of Fisheries Education,
Versova, Mumbai, 400061, India.
E-mails: ashutoshddeo@gmail.com;
ashutosh@cife.edu.in
Funding information
Indian Council of Agricultural Research
(ICAR)
Abstract
A 60-day feeding trial was conducted to determine the growth performance and
expression of insulin-like growth factor 1 gene (IGF-I gene) in Labeo rohita fingerlings
fed with either raw, soaked or fermented sweet potato leaf meal (SPLM) by com-
pletely replacing de-oiled rice bran (DORB), following a completely randomized
design. Seven isonitrogenous (30%) and isocaloric (1.8 MJ/100 g) diets were pre-
pared by replacing DORB with 50% and 100% raw, soaked and fermented sweet
potato leaf meal, maintaining DORB-containing diets as a control. Weight gain %,
SGR (specific growth rate) and PER (protein efficiency ratio) were significantly
(p < 0.05) higher when 100% DORB was replaced by fermented SPLM in compar-
ison to other treatment groups. The fermented and soaked SPLM-fed groups had
registered with lower FCR value. The expression of growth regulating gene IGF-I
mRNA and RNA/DNA ratio was found to be significantly higher (p < 0.05) in soaked
and fermented SPLM-fed groups. In this study, the body protein and lipid composi-
tion did not vary significantly (p > 0.05). Hence, the study concludes that the fer-
mented sweet potato leaf meal using Chaetomium globosum can replace 100% DORB
in the diet of Labeo rohita without any detrimental effect on growth performance.

KEYWORDS
de-oiled rice bran, growth performance, insulin-like growth factor 1 gene, Labeo rohita, RNA/
DNA ratio

1 | INTRODUCTION
Fish farming is the fastest growing agricultural sector in the world
(SOFIA, 2016). The increasing human population has a lead increased
demand for quality protein. Fish being a good protein source has
witnessed an increase in per capita apparent fish consumption from
an average of 9.9 kg in the 1960s to 20 kg in 2016 (SOFIA, 2016).
India is dominating in aquaculture production, which includes many
species like carps, catfishes, shellfish, pangasius, tilapia etc. Among
the three Indian major carps (IMC), Labeo rohita is the most
important cultivable species and high consumer demand due to its
taste preference. The aquaculture has also grown rapidly and led to
more feed based culture system. Feeding is the most important fac-
tors required for faster growth and higher yield of cultured Labeo
rohita (Jose, Mohan, Shyama, Ramachandran Nair, & Mathew, 2006;
Maity & Patra, 2008). Aquafeed ingredients mainly depend on oil
cake as the main conventional protein sources and de-oiled rice bran
(DORB) as an energy source for the carp culture in the Asian sub-
continent. As a practice, DORB (De-Oiled Rice Bran) is most com-
monly used feed ingredient for carp culture. In carp culture, DORB is

Aquaculture Research. 2018;1–10. wileyonlinelibrary.com/journal/are © 2018 John Wiley & Sons Ltd | 1
2 |
used either as mash feed or a combination of mash and pelleted
feed (Singh, Garg, Bhatnagar, & Kalla, 2004). DORB is cheapest agri-
cultural by-product available throughout the year which is predomi-
nantly used along with oil cakes in the feed formulation and well
accepted by the carps (Veerina, Nandeesha, & Gopal Rao, 1993).
DORB is fat-free rice bran or rice polish and it is a good replacer of
other aquafeed ingredients of low protein (<20%) as energy source
(Limbu et al., 2016). Using higher level of DORB in aquafeed the
cost of aquafeed can be reduced substantially (Ramakrishna, Shipton,
& Hasan, 2013). However, it has been observed in the recent past
that the price of DORB has increased substantially. Therefore, there
is an urgent need to look for the potential alternate ingredient to
DORB that must be available in plenty.
There are plenty of other potential ingredients available locally,
however, irregular supply and presence of antinutritional factors make
it difficult to include those ingredients in aquafeed preparations. In
the present scenario, leaf meals are one of the cheapest sources of
protein and energy that may be a potential alternative in future and
will reduce the cost of fish feed (Adewolu, 2008). Leaf meal may be
the best option to replace protein and energy sources for animal feed
but their nutritive and antinutritive potentials are yet to be ade-
quately studied and utilized (Osman, 2007; Vhanalakar & Muley,
2014). Among such plants, Ipomoea batatas leaf serves as an effective
protein and energy source for the formulation of fish diets. Several
studies have been conducted to estimate the nutritive value of sweet
potato leaf meal viz. Adewolu (2008); Ishida et al. (2000). The sweet
potato leaf meal (SPLM) has high protein content 23%–33% with the
high amino acid score in addition to the substantial amount of nutri-
ents; therefore, it should be recommended in the diets of animals
(Antia, Akpanz, Okonl, & Umorenl, 2006). However, the major factor
limiting the use of SPLM in animal feed is the presence of antinutri-
tional factors which reduces its digestibility and hampers the growth.
Sweet potato leaf contains phytate, trypsin inhibitor, alkaloid, oxalate,
tannin and cyanide as antinutritional factors (Antia et al., 2006).
Several methods have been used and documented to remove the
ANFs from SPLM (Campbell & Bedford, 1992; Mwachireya, Beames,
Higgs, & Dosanjh, 1999; Almazan, 1995). Soaking of ingredients
helps in reduction in antinutritional factors; however, fermentation
was reported to be an innovative approach by which nutrient
digestibility and their availability can be increased (Kim, Yang, &
Song, 1999). In recent year several studies have been conducted on
the use of fermented ingredients in the diets of monogastric animals
such as chicken (Hirabayashi, Matsui, & Yano, 1998), piglets (Kiers
et al., 2003) and also in fish feed (Yamamoto et al., 2010; Yuan et al.,
2013). The biological detoxification of agro-industrial and plant ingre-
dients has been the main focus of solid-state fermentation (SSF)
research, with the basic aim to improve the nutrients status and its
bioavailability in animals by reducing the ANFs. Improve the nutrient
utilization is the bidirectional approach of one is the processing mod-
ification for reducing the ANF and increasing digestibility.
Nutrigenomics approach is a new advancement in nutritional
research understanding the nutrition and genomics effect of diets on
gene expression pattern (Mutch, Wahli, & Williamson, 2005). During
MESHRAM ET AL.
the last few years, the researchers are more focusing on molecular
research to verify the data generated from the traditional feeding
trial. The molecular tools help us to evaluate particular ingredients or
feed performance in particular species through metabolic markers.
Insulin-like growth factor (IGF) axis is a marker reported to be asso-
ciated with growth and development (Jones & Clemmons, 1995;
Kumar, Sandor Zs, et al., 2017; Kumar, Sahu, et al., 2017). Insulin-
like growth factor I are mitogenic peptides, which are regulated by
the nutritional status of fish and also have a significant role in nutri-
ent metabolism and growth (Duan, 1998; Tatar, Bartke, & Antebi,
2003). Insulin-like growth factors are produced mainly in the liver.
Insulin-like growth factor 1 has been shown the wide range of bio-
logical function including cell division, differentiation and also growth
(Jones & Clemmons, 1995). The insulin-like growth factor plays
important role in the development and somatic growth in verte-
brates (Picha, Turano, Beckman, & Borski, 2008). Therefore, the
objective of present study was to evaluate the growth performance
and expression of IGF-I gene in L. rohita fed with raw, soaked and
fermented sweet potato leaf meal by replacing DORB.
2 | MATERIALS AND METHODS
2.1 | Preparation of SPLM
Prior to incorporation in the diets, SPLM was given different treat-
ments with water soaking or solid state fermentation for removal of
antinutritional factors. For soaking 20 g leaf sample was taken in fine
clothbound triplicates in beakers (500 ml vol.) and added 200 ml of
tap water to simulate field condition of future use. Soaking was
done up to 24 hr; at every 4-hr interval, water was exchanged. After
soaking the leaf was oven dried at 60°C for overnight. The soaked
dried sample was milled and pass through 40 l mesh sieves and
stored in airtight container. Solid-state fermentation (SSF) of sweet
potato leaf meal was carried out using a fungus Chaetomium globo-
sum (MTCC-4179) for 120 hr based on the result of our earlier labo-
ratory work for maximum protein. About 20 g of dried sweet potato
leaf meal (particle size < 40 l) was added into the conical flask in
triplicate. Based on the standardization results, moisture content was
adjusted at 50% using distilled water and then it was autoclaved for
15 min at 121°C at 1 kg/cm2 pressure, after which they were
allowed to cool. After that, the sterilized SPLM samples were inocu-
lated with C. globosum (3 9 105 cell/g) and mixed properly for uni-
form distribution of inoculate. In addition to these, untreated sweet
potato leaf meal was used as a raw sample.
2.2 | Experimental setup
The experiment was conducted for a period of 60 days in the wet
laboratory of Central Institute of Fisheries Education, Mumbai. The
setup consisted of 21 plastic rectangular tubs (57 9 36 9 47 cm,
100 L capacity) covered with perforated lids. Two hundred ten fishes
(10 fish in each tank of ave.wt.7.2
0.05 g) were equally dis-
tributed in seven distinct experimental groups in triplicate following
MESHRAM ET AL. | 3

a completely randomized design. Feeding was done up to satiation
twice daily until the end of the experiment. The aeration pipe in
each tub provided with an air stone and a control valve to regulate
the air pressure uniformly. The water quality parameter viz. tempera-
ture, dissolved oxygen pH and ammonia were recorded and main-
tained within the optimum range (Temperature 29–30°C; pH 7.5–
8.2; DO 6.3–7.5 mg/L and ammonia 0.05–0.08 ppm) during the
experimental period. Siphoning was done daily to remove the excess
excreta and residual feed. The same volume of water was replen-
ished after siphoning.
replacement), TR100 (treatment with 100% raw SPLM in replace-
ment), TS50 (treatment with 50% soaked SPLM in replacement),
TS100 (treatment with 100% soaked SPLM in replacement), TF50
(treatment with 50% fermented SPLM in replacement) and TF100
(treatment with 100% fermented SPLM in replacement). Practical
ingredients such as groundnut oil cake, de-oiled rice bran, wheat
flour, carboxymethyl cellulose (CMC), defatted soybean meal, buty-
lated hydroxytoluene (BHT), betaine, cod liver oil, sunflower oil vita-
min mix and mineral mixture, were taken for feed preparation. The C
(control) had only 30% de-oiled rice bran.

2.3 | Diet preparation 2.4 | Proximate composition and antinutritional

Seven isonitrogenous (30% CP) and isocaloric (1.8 MJ/100 g) practi-
cal diets were prepared (Table 1) with the replacement of de-oiled
rice bran with raw (R), soaked (S) and fermented (F) SPLM viz. C
(control with DORB), TR50 (treatment with 50% raw SPLM in
factor
Proximate composition of the experimental diets and fish were anal-
ysed following AOAC (1995). The experimental diets and body sam-
ples were dried at 102
2°C in a hot air oven to determine the

T A B L E 1 Feed and proximate composition of the experimental diets (DM basis) for feeding trial
Ingredients C TR50 TR100 TS50 TS100 TF50 TF100 SPLM DORB
GNOC 29.5 25.375 21.1 25.25 22.2 22.7 16.25
RSPLM 0 15 30 0 0 0 0
SSPLM 0 0 0 15 30 0 0
FSPLM 0 0 0 0 0 15 30
DORB 30 15 0 15 0 15 0
DSBM 25 25 25 25 25 25 25
Wheat flour 6.625 10.75 15.025 10.875 13.925 13.425 19.875
Sunflower oil 3 3 3 3 3 3 3
Fish oil 3 3 3 3 3 3 3
Vit–min mix 2 2 2 2 2 2 2
Vit C 0.1 0.1 0.1 0.1 0.1 0.1 0.1
BHT 0.025 0.025 0.025 0.025 0.025 0.025 0.025
CMC 0.5 0.5 0.5 0.5 0.5 0.5 0.5
Betaine 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Total 100 100 100 100 100 100 100
Proximate composition
Dry matter (%) 91.6 93.1 92.0 92.3 91.8 92.3 91.8 95.31
0.30 90.26
0.22
Crude Protein (%) 30.8 29.8 30.6 30.6 29.8 30.7 29.9 23.91
0.817 14.57
0.05
Crude lipid (%) 6.57 7.14 7.23 7.12 7.27 7.28 7.32 5.55
0.55 0.26
0.04
Ash (%) 9.61 10.17 9.82 10.28 9.99 10.74 10.24 10.17
0.84 5.62
0.08
Crude fibre (%) 9.98 10.25 9.40 9.6 9.52 9.60 8.54 8.3
0.16 14.85
0.19
NFE (%) 43.0 42.6 42.8 42.3 43.3 41.5 43.9 52.04
0.57 54.95
0.38
GE(MJ/100 g) 1.81 1.80 1.81 1.81 1.82 1.83 1.81

Note. BHT, butylated hydroxytoluene; C, control; CF, crude fibre; CL, crude lipid; CMC, carboxymethyl cellulose; CP, crude protein; DORB, de-oiled rice
bran; DSBM, defatted soybean meal; F, fermented; FSPLM, fermented sweet potato leaf meal; GE, gross energy, gross energy expressed in terms of
MJ/100 g; GNOC, groundnut oil cake; NFE, nitrogen-free extract.(NFE = 100(CP + CL + CF + ash); R, raw; RSPLM, raw sweet potato leaf meal; S,
soaked; SSPLM, soaked sweet potato leaf meal; T, Treatment.
Vit–min mix-vitamin and mineral mixture. Composition of vitamin mineral mix (PREEMIX PLUS) (quantity/kg): Vitamin A, 55,00,000 IU; Vitamin D3,
11,00,000 IU; Vitamin B2, 2,000 mg; Vitamin E, 750 mg; Vitamin K, 1,000 mg; Vitamin B6, 1,000 mg; Vitamin B12, 6 mcg; Calcium Pantothenate,
2,500 mg; Nicotinamide, 10 g; Choline Chloride, 150 g; Mn, 27,000 mg; I, 1,000 mg; Fe, 7,500 mg; Zn, 5,000 mg; Cu, 2,000 mg; Co, 450 L- lysine,
10 g; DL- Methionine, 10 g; Selenium.
All data are expressed in the terms of Mean
SE in triplicates, values with common superscripts letters in each row are significantly different
(p < 0.05).
4 | MESHRAM ET AL.

moisture content. The crude protein content of the raw material, into desiccators to cool and then reweighed until obtained a con-
diets and body sample was estimated using micro-Kjeldahl methods stant weight. The weight of the alkaloid was determined by the
(Kelplus, PELICAN, India) crude lipid was determined by Soxhlet weight difference of the filter paper.
extraction method (SOCS plus, SAS-AS 08 PELICAN, India). Fibre The alkaline titration method of AOAC (1984) was used for the
estimation was measured using fibre tech (Tulin equipment’s, India) determination of hydrogen cyanide. About 5 gram of finally ground
and the ash content was done using a muffle furnace at 550°C for sample was taken in 100 ml of distilled water and kept at room
5 hr. temperature for 2 hr. The solution was filtered and steam-distilled
The antinutritional factors present in SPLM (raw, soaked and fer- with NaOH solution. The distillate was treated with diluted KI solu-
mented) were determined by various methods. The phytic acid in tion. The solution was titrated against 0.02M AgNO3 solution. The
the extract was determined according to the colorimetric assay endpoint was obtained when there was a change from clear to a
described by Gao et al. (2007). 0.5 g defatted dried SPLM was taken faint but permanent turbid solution. Hydrogen cyanide was deter-
in 100-ml Erlenmeyer flask and extracted with 10 ml of 3.5% HCl. mined by taking 1 ml of 0.02 M AgNo3 equivalent to 1.08 mg
The samples were constantly shaken for 1 hr in the shaker HCN.
(ORBITEKâ; Scigenics, India) at 200 rpm followed by 10 min cen-
trifugation at 1,677 g and collected the supernatant. The sample
2.5 | Growth parameters
(0.1 ml) extract was diluted with 3 ml distilled water and mixed with
1 ml Wade reagent (0.03% FeCl36H2O + 0.3% sulphosalicylic acid) The growth performance such as weight gain, specific growth rate
and the mixture was centrifuged at 1,677 g for 10 min. After 30 min (SGR), feed conversion ratio (FCR) and protein efficiency ratio (PER)
the supernatant was taken for absorbance at 500 nm against dis- of fish were evaluated at the end of feeding trial using following
tilled water. The phytic acid was calculated from the calibration equations.
curve of phytic acid standard.
For the estimation of tannin, the spectrophotometric method as
Weight gainð%Þ ¼ Final weightðgÞ
described by Makkar, Siddhuraju and Becker (2007) was followed.
 Initial weightðgÞ=Initial weightðgÞ  100
Folin– Ciocalteu reagent was used in this method and the results
were expressed as the tannic acid equivalents. The total tannin
SGR ¼ ðLoge Final weight
content was calculated from the calibration curve of tannic acid
 Loge Initial weightÞ=ðnumber of daysÞ  100
standard.
Oxalate was estimated following the method of Nwosu (2011).
FCR ¼ Feed givenðg dry weightÞ=Body weight gainðg wet weightÞ
One gram sample was taken in 100 ml beaker, added 20 ml of
 100
0.03N HCl, warmed for 1 hr at 40–50°C using a magnetic hot plate
and stirred. The extract was diluted to 100 ml in a volumetric flask. PER ¼ Net weight gain=protein fedðgÞ  100
About 5ml of the extract was taken in a conical flask and made alka-
line by adding 1 ml of 5N ammonium hydroxide. Then it was made
2.6 | Body indices
acidic by adding glacial acetic acid. 1 ml of 5% CaCl2 and 3 drops of
phenolphthalein and allowed to stand for 3 hr and centrifuged for
2.6.1 | Hepatosomatic index (HSI)
15 min at 604 g. The supernatant was discarded and 2 ml of 3N
H2SO4 was added to each tube and the precipitate was dissolved by The liver weights of fish of different treatment groups were
warming at 70°C. The contents of all the tubes were carefully recorded and the Hepatosomatic index was calculated as follows:
poured into a clean conical flask and titrated with 0.01N KMNO4
HSI ¼ Liver WeightðgÞ=Weight of fishðgÞ  100
freshly prepared at room temperature until the first pink colour.
Then, it was allowed to stand until the solution became colourless.
The solution was warmed at 70°C and titrated until a permanent 2.6.2 | Intestinal somatic index (ISI)
pink colour appeared for at least 30 s.
The weights of gastrointestinal tract of fish from different treatment
For the estimation of alkaloids 5 g of sample was weighed in
groups were recorded and the intestinal somatic index was calcu-
triplicates following gravimetric method suggested by Harborne
lated as follows:
(1973). The sample was dispersed into 50 ml of 10% acetic acid
solution in ethanol. Before filtering the mixture was shaken and then ISI ¼ Intestine weight ðgÞ=weight of fishðgÞ  100
allowed to stand for 4 hr. The filtrate was then evaporated to one-
quarter of its original volume. Concentrated ammonium hydroxide
2.7 | Quantification of DNA, RNA and RNA: DNA
was added drop wise in order to precipitate the alkaloids. A pre-
ratio
weighed filter paper was used to filter off the precipitate and it was
then washed with ammonium hydroxide solution (1%). The precipi- Determination of nucleic acids in tissue was performed by pentose
tate filter paper was dried at 60°C for 30 min in oven, transferred analysis following the method of Schneider, Calowick and Kaplon
MESHRAM ET AL.
(1957) and was calculated as follows: The DNA content of the
nucleic acid extract is presented by the following equation:
lgDNA mL1 ¼ 1=4 OD at 600nm  0:019
The RNA content of the nucleic acid extract is presented by the
following equation:
lgRNA mL1 ¼ ½ðOD at 660nm þ 0:0081Þ
 ðlgDNA mL1  0:013Þ=0:116
2.8 | RNA extraction and cDNA synthesis
Approximately 100 mg of liver tissue was mechanically homogenized
with 1 ml of the TRI reagent (Sigma, St. Louis, MO, USA) and the
total RNA was extracted according to the manufacturer’s instruc-
tions. The purity and concentration of RNA were checked in Nano-
Drop spectrophotometer (Thermoscientific, USA). The RNA samples
were treated with DNase (Fermentas, USA) according to the proto-
col for the removal of residual DNA during preparation. cDNA was
synthesized from the 1 lg of total RNA using oligo dT primer follow-
ing the protocol of first stand cDNA synthesis kit (Fermentas, USA).
2.9 | Quantitative RT-PCR
The relative quantification of IGF-I mRNA expression was performed
using AriaMx Real-Time PCR System (Agilent Technology, LDA
Malaysia) using SYBER Green qPCR master mix without ROX (Ther-
moscientific, USA). Standard reaction mixtures (10 µl) were assem-
bled using 5 µl of SYBER Green supermix 29, 1 µl each primer
(5 pm) and 100 ng of template cDNA For amplification of IGF-I
gene-specific primer and beta-actin reference gene primer (Table 2)
were designed using Gene runner software (version 3.05) based on
sequences available in GenBank (http://www.ncbi.nlm.nih.gov). The
melting curve analysis of the target gene and reference gene
resulted in single products with specific melting temperature. Com-
parative CT values were used to estimate the relative expression of
mRNA. All reaction run in duplicates and the data were analysed
using the 2DDct method (Livak & Schmittgen, 2001).
2.10 | Statistical analysis
All values presented are the mean
standard error. Statistical analy-
sis of data was performed by analysis of variance (ANOVA) in SPSS
(Version 22) followed by Duncan’s multiple range test. Significance
was considered at p < 0.05.
T A B L E 2 Primer used for
Gene Accession no.
expression study in real time PCR
IGF-I KX455870
b-actin EU184877
| 5
3 | RESULTS
3.1 | Proximate composition of diets and fish body
Proximate composition of diets and SPLM, DORB are presented in
Table 1. The crude protein and lipid content in fermented SPLM
increased from 23.92% and 5.55% to 30.34%, and 7.05% respec-
tively but the crude fibre decreased from 8.30% to 6.01% (Table 3).
After 24 hr soaking, the protein content decreased from 23.92% to
23.27%, crude lipid from 5.55% to 5.14% and fibre content change
from 8.30% to 8.10% (Table 3). The whole body compositions of all
the experimental fish of different treatments are presented in
Table 4. Dietary replacement of de-oiled rice bran by SPLM did not
affect (p > 0.05) the moisture, crude protein and lipid. However, the
ash content was significantly higher in TF100 which is similar to
TF50. The crude protein content varied from 14.8% to 16.4%,
whereas lipid content of all the fish was estimated within 3.64% to
4.3% and total ash content varied from 3.2% to 4.88%. The reduc-
tion in antinutritional factor (phytate, oxalate, alkaloid, tannin and
HCN) after soaking and fermentation are presented in Table 3.
3.2 | Growth performance
There was significant difference (p < 0.05) in the body weight gain
among the different treatment groups at the end of the experiment
(Table 5). The weight gain % specific growth rate and PER in TF100
were significantly (p < 0.05) higher as compared to other treatment
groups. However, the highest FCR was recorded in TR100 group,
which was similar to TR50. The lower FCR was recorded in TF50-
and TF100-fed group which is statically similar to TS50- and TS100-
fed group.
3.3 | Body indices
The HSI value of different treatment was not significantly different
(p > 0.05) (Table 5) among the treatment groups but the ISI value of
different experimental groups varied significantly (p < 0.05). In this
study, the ISI value in TF50 fed group was significantly (p < 0.05)
higher and lowest value was recorded in control-fed group.
3.4 | RNA, DNA and RNA/DNA ratio
The RNA and RNA/DNA ratio varied significantly (p < 0.05) in the
different treatment groups (Table 6). The DNA content did not show
Primer sequence Amplicon size Anneline temp.
F-GCAAACCGACAGGCTATGGGC 166 57
R-GTGTCTGTGTGCCGTTCCGC
F-CACTGCTGCTTCCTCCTCCTCC 138 58
R-GATACCGCAAGACTCCATACCCAAG
6 | MESHRAM ET AL.

T A B L E 3 Effect of soaking
Treatment Particulars Raw(0 hr) Soaked(24 hr) Fermented(120 hr)
(24 hr) and solid-state
Proximate composition CP (%) 23.9b
0.1 23.4b
0.42 30.3a
0.48 fermentation (120 hr) on proximate
CL (%) 5.55
0.68 5.14
0.52 7.05
0.41 composition and antinutritional
CF (%) 8.30
0.19 8.10
0.19 6.01
0.75 factors of SPLM

ASH (%) 10.17
1.01 9.60
1.45 10.21
1.45

NFE (%) 52.04
0.96 53.86
1.35 46.36
2.10
Antinutritional factor Tannin (mg/100 g) 23.03
0.94
a
4.84
0.56
c
8.53b
0.14
Try (TIA mg/g 3.20
0.46 2.50
0.22 2.90
0.16
Phy (mg/100 g) 15.02
0.68
a
11.06
0.05
b
9.38c
0.10
Oxalate (%) 1.36
0.01
a
0.29
0.007
c
0.94b
0.01
Alkaloid (mg/g) 1.36a
0.14 1.00b
0.17 0.73b
0.06
HCN (mg/100 g) 38.8
1.24
a
23.0
1.90
b
23.04b
1.9

Note. CF, crude fibre; CLcrude lipid; CP, crude Protein; HCN, hydrogen cyanide; NFE, nitrogen-free
extract; Phy, phytate; Try, trypsin inhibitor.
Data represent mean
S.E. Values in the same column with different superscripts are significantly differ-
ent (p < 0.05), n = 3.

T A B L E 4 Proximate
Treatment Moisture CP CL TC Ash
composition of whole body of
C 73.7
0.90 16.4
0.18 4.04
0.38 1.66
0.66 4.12abc
0.31 Labeo rohita fingerlings of different
TR50 72.8
1.14 15.8
0.66 4.31
0.37 3.41
0.25 3.60bc
0.06 experimental groups (%wet weight
TR100 73.9
1.39 15.5
1.02 4.08
0.49 3.09
0.46 3.24c
0.16 basis)

TS50 73.9
0.94 15.3
0.62 4.01
0.30 2.38
0.46 4.34abc
0.48
TS100 74.2
2.25 15.5
1.25 4.07
0.69 1.49
0.24 4.61ab
0.07
TF50 75.1
1.51 14.8
1.14 3.64
0.44 1.51
0.71 4.85a
0.50
TF100 74.1
1.05 15.3
1.78 3.99
0.62 1.65
1.04 4.88a
0.31

Note. CL, crude lipid; CP, crude protein; TC, total carbohydrate (TC = 100(%moisture + %CP + %CL +
%ash).
Data represent mean
S.E. Values in the same column with different superscripts are significantly differ-
ent (p < 0.05) n = 3.

any significant (p > 0.05) variation among the treatment groups.
However, the RNA concentration and RNA/DNA ratio was found
significantly higher in the treated SPLM fed groups compared to the
control group and raw SPLM-fed groups. The RNA concentration
was found significantly higher in TS50-fed group followed by TS100
treatment groups. RNA/DNA ratio was observed significantly higher
in TF100-fed groups.
3.5 | Expression of IGF-I mRNA of Labeo rohita
liver tissue
The expression of IGF-I in liver tissue of L. rohita varied significantly
(p < 0.05) among the treatment groups (Figure 1). IGF-I mRNA
expression was significantly (p < 0.05) higher in TF100 and TS100,
while control, TR50- and TR100-fed groups showed lower value
(Figure 1).
4 | DISCUSSION
SPLM is important source of protein and energy for fish feed but some
antinutritional factor reduced the nutritional value and inhibited the
growth performance of animal. Therefore, to increase SPLM nutri-
tional value two method, microbial fermentation and water soaking
was planned. It was for replacing the expensive protein resources by
SPLM to make cost effective feed for animal feed production. Reduc-
tion in antinutritional factors (phytate, tannin, alkaloid, HCN and oxa-
late) from plant ingredients for sustainability have been taken by many
researchers (Kumar, Makkar, Amselgruber, & Becker, 2010; Makkar &
Becker, 2008; Shamna et al., 2015). In the present study increased
nutritional values have been observed with reduction in antinutritional
factors. Similar to earlier studies (Vijayakumari, Siddhuraju, Pugalenthi,
& Janardhanan, 1998; Nwosu, 2010), soaking increased the nutritional
value of SPLM by decreasing the antinutritional factors (Table 3).
Solid-state fermentation is also important and an effective method for
reducing antinutritional factors to improve nutritional value (Yuan
et al., 2013). In the present study, crude protein and lipid content in
fermented SPLM increased from 23.92% and 5.55% to 30.34%, and
7.05% respectively and the crude fibre decreased from 8.30% to
6.01%. It gave a very considerable change in nutritional value improve-
ment. Previous studies focused on the replacement of ingredients in
the feed, based on the growth data of the experimental animal. Soak-
ing and solid-state fermentation (SSF) are well-documented for the
improvement of ingredients quality. According to Nwosu (2010),
MESHRAM ET AL. | 7

T A B L E 5 Growth performance of Labeo rohita fingerling fed with different experimental diets
Treatment WG% SGR FCR PER HSI ISI
C 84.5c
4.01 1.02c
0.03 2.03bc
0.03 1.59bc
0.03 1.04
0.048 2.21b
0.23
TR50 83.0
3.54
c
1.00
0.03
c
2.17 ab

0.04 1.53 cd

0.03 0.93
0.04 2.90b
0.16
TR100 81.6c
2.19 0.99c
0.02 2.24a
0.09 1.45d
0.02 0.90
0.05 3.22ab
0.19
TS50 88.8 bc

2.48 1.05 bc

0.02 1.93 cd

0.06 1.69 ab

0.06 1.01
0.08 3.03b
0.49
TS100 90.5 abc

1.66 1.07 abc

0.01 1.88 cd

0.03 1.77
0.03
a
1.09
0.24 2.78b
0.57
TF50 94.8ab
1.97 1.11ab
0.01 1.82d
0.06 1.78a
0.06 0.97
0.13 4.30a
0.67
TF100 98.1
2.45
a
1.13
0.02
a
1.79
0.04
d
1.82
0.04
a
1.19
0.23 2.71b
0.22
p-value 0.005 0.006 0.000 0.000 0.833 0.037

Note. FCR, feed conversion ratio; FER, feed efficiency ratio; HSI, hepatosomatic index; ISI, intestinal somatic index; PER, protein efficiency ratio; SGR,
specific growth rate; WG, weight gain.
Data represent mean
S.E. Values in the same column with different superscripts are significantly different (p < 0.05) n = 3.

T A B L E 6 DNA, RNA and RNA/DNA ratio in the muscle of Labeo and enhances the growth performance in the animal (Ramachandran,
rohita fingerling fed with different experimental diets Bairagi, & Ray, 2005).
Treatments lg DNA/ml lg RNA/ml RNA/DNA ratio In the present study, an effort was made to evaluate the replace-
C 11.3
0.30 1.03
0.17
c
0.09 bc

0.01 ment of de-oiled rice bran (DORB) with raw sweet potato leaf meal,
TR50 10.5
0.54 0.89c
0.29 0.08c
0.02 soaked sweet potato leaf meal and fermented sweet potato leaf

TR100 11.0
0.70 1.10bc
0.06 0.10bc
0.1

meal in Labeo rohita diet. It was observed that the replacement of
DORB with soaked and fermented SPLM significantly increased the
TS50 12.8
0.33 1.65
0.16
a
0.12 abc

0.01
growth performance and feed utilization as compared to raw SPLM.
TS100 12.1
1.0 1.60ab
0.04 0.13ab
0.01
The TF100 (fermented with 100% replacement of DORB) had higher
TF50 10.4
0.69 1.54a
0.08 0.14a
0.01
WG% and SGR as compared to control and other treatment groups.
TF100 10.3
0.61 1.53ab
0.03 0.15a
0.01
Hassan, Soltan and Abdel-Moez (2015) showed that up to 37.7%
p value 0.115 0.011 0.023
fish meal in diets of tilapia could be replaced by yeast-fermented
Note. C, control; DNA, Deoxyribonucleic acid; RNA, Ribonucleic acid; T, soyabean meal without affecting the growth performance and nutri-
Treatment.
ent utilization. It was also reported that 30% fermented (intestinal
Data represent mean
S.E. Values in the same column with different
superscripts are significantly different (p < 0.05) n = 3. bacterium) Lemna leaf meal resulted in the best growth and feed uti-
lization efficiencies in rohu fingerlings (Bairagi, Ghosh, Sen, & Ray,
2002). They also found that fish fed fermented leaf meal-containing
1.4 diets were superior in growth to those fed diets containing raw leaf

1.2
IGF-I a meal (Bairagi et al., 2002). Felix and Brindo (2013) had reported that
the fish meal can be substituted with fermented and raw seaweed
ab
1 (Kappaphycus alvarezzii) in the diet of Macrobrachium rosenbergii up
Fold change

0.8 to 20% and 30% respectively. The present result of better growth
bc performance in 100% fermented SPLM fed group is being supported
bc
0.6
by Wang et al. (2017). They reported feeding SSF feed could signifi-
c c c
0.4 cantly improve the growth performance and digestion in sea cucum-
ber nursery and grow-out phase. The nutrient level was significantly
0.2
increased by the fermentation with Chaetomium globosum. The
0 improvement of nutritional value of fermented plant-based ingredi-
C TR50 TR100 TS50 TS100 TF50 TF100
ents have been reported by many authors viz., sesame seed meal
Treatments
(Bairagi et al., 2002); palm kernel meal (Ng & Wee, 1989) and cot-
F I G U R E 1 Expression of IGF-I mRNA Labeo rohita liver tissue at tonseed meal (Sun et al., 2015), which corroborated our findings.
different experimental groups TS100-fed group had higher WG% and SGR compared to control
and TR100(raw SPLM replacing 100% DORB)-fed group. Soaked
sweet potato leaf meal had significantly positive effect on growth
among the pre-processing method soaking is the effective method for performance and nutrient utilization in L. rohita, which is in agree-
the reduction in the antinutrient factors. Similarly, solid-state fermen- ment with Ndau and Madalla (2015), who found that the effect of
tation has been reported to decrease the antinutritional factors, thus soaked pigeon pea seed meal with inclusion level up to 45% could
enhances the utilization of nutrients by improving nutrients quality, replace fish meal with better growth performance in Nile tilapia.
8 |
Olude, Alegbeleye and Obasa (2008) found soaked copra meal can
be incorporated in the diet of Nile tilapia up to 30% without any
deleterious effect on growth performance and nutrient utilization. It
is important to mention that in the present study, the DORB was
replaced with raw and processed sweet potato leaf meal which is
first attempts of this kind hence; in the absence of authentic pub-
lished literature, it could not be compared or discussed in particular
to carp feed.
Several studies considered RNA/DNA ratio as an index of
growth (Akhtar, Pal, Sahu, Ciji, & Gupta, 2012; Gupta, Pal, Sahu, Jha,
& Kumar, 2015; Kumar, Sahu, & Ranjan, 2018; Kumar, Sandor Zs,
et al., 2017; Kumar, Sahu, et al., 2017). The quantitative analysis of
RNA and DNA can disclose the growth rate in fish as RNA is an
organizer of protein synthesis. The somatic growths increased with
increased RNA content (Bulow, 1987) while the DNA content
remains constant. RNA is organizer molecule for protein synthesis.
The RNA/DNA ratio is a sensitive index of anabolic activity and
highly varies depending on the nutritional status and feeding (Tanaka
et al., 2007). The present study indicated that the RNA concentra-
tion and RNA/DNA ratio were significantly higher in soaked and
solid state fermented SPLM replacing DORB-fed group as compared
to control and raw. Similarly, the RNA/DNA ratio indicates the
growth performance in fish L. rohita (Akhtar et al., 2012; Kumar,
Sahu, & Pal, 2006; Kumar, Sandor Zs, et al., 2017; Kumar, Sahu,
et al., 2017).
As the growth of animal is affected by many factors, evaluation
of growth only by considering weight gain of the animal due to feed-
ing of particular ingredients may not be justified. Hence, molecular
responses of the growth-regulated gene are being used to choose
for evaluation of particular ingredients in the feed of a targeted spe-
cies. The GH/IGF-I axis provides an integral signal for growth and
nutrient partitioning (Beckman & Dickhoff, 1998) and it is also
involved in metabolism and tissue differentiation. Insulin-like growth
factors (IGF), IGF-I belong to the growth regulating family. These
hormones are synthesized in liver and its expression is controlled by
the quantity of feed fed and its nutrient composition in fish and
birds (Clemmons & Underwood, 1991). The expression of IGF-I con-
centration was positively correlated with the growth rate in Salmo
salar (Dyer et al., 2004). Dietary protein level alters the expression
of IGF-I in fish (Gomez-Requeni et al., 2004) and also interacts with
nutritional status of fish (Margaret, Cheng, & Chan, 2006). Insulin-
like growth factor I axis is regulated by the nutritional and metabolic
condition. Its subsequent effect on liver and liver for modulating
sensitivity to growth hormone is a real concern (Pierce, Breves, Mor-
iyama, Hirano, & Grau, 2011). In the present study highest expres-
sion (p < 0.05) of IGF-I mRNA was observed in TF100-fed group.
Similar observation has been made by Kumar, Sandor Zs, et al.,
2017; Kumar, Sahu, et al., 2017 who reported decreased growth
performance with decreased IGF-I gene expression in Silurus glanis
when fishmeal was replaced with soyabean meal. The IGF-I mRNA
expression is an important indicator of growth in Tilapia (Uchida
et al., 2003); Labeo rohita (Kumar, Sandor Zs, et al., 2017; Kumar,
Sahu, et al., 2017).
MESHRAM ET AL.
5 | CONCLUSION
The present study clearly established that the SPLM has potential as
aquafeed ingredients. The fermented sweet potato leaf meal using
Chaetomium globosum can replace 100% de-oiled rice bran in the
diet of L. rohita without any detrimental effect on growth perfor-
mance, nutrient utilization which was also verified with the expres-
sion of growth gene IGF-I. However, further study needs to be
conducted on economic feasibility of SPLM as a replacer of DORB
and also to evaluate the long-term feeding effect of fermented
sweet potato leaf meal on the growth and other related parameters
of fish before SPLM is recommended for commercial application in
the carp diets.
ACKNOWLEDGMENTS
The Authors are thankful to Indian Council of Agricultural Research
(ICAR) for the financial support. Director of ICAR- CIFE (Indian
Council of Agricultural Research-Central Institute of Fisheries Educa-
tion) for providing the necessary infrastructural facilities for complet-
ing the experiment.
CONFLICT OF INTEREST
The authors declare no competing financial interests.
ORCID
Ashutosh D Deo http://orcid.org/0000-0002-9341-2084
Narottam P Sahu http://orcid.org/0000-0003-3932-7302
REFERENCES
Adewolu, M. A. (2008). Potentials of sweet potato (Ipomoea batatas) leaf
meal as dietary ingredient for Tilapia zilli fingerlings. Pakistan Journal
of Nutrition, 7(3), 444–449. https://doi.org/10.3923/pjn.2008.444.
449
Akhtar, M. S., Pal, A. K., Sahu, N. P., Ciji, A., & Gupta, S. K. (2012). Effects
of dietary pyridoxine on growth and biochemical responses of Labeo
rohita fingerlings exposed to endosulfan. Pesticide biochemistry and
physiology, 103(1), 23–30. https://doi.org/10.1016/j.pestbp.2012.02.
004
Almazan, A. M. (1995). Antinutritional factors in sweet potato greens.
Journal of Food Composition and Analysis, 8(4), 363–368. https://doi.
org/10.1006/jfca.1995.1031
Antia, B. S., Akpanz, E. J., Okonl, P. A., & Umorenl, I. U. (2006). Nutritive
and Anti-Nutritive Evaluation of Sweet Potatoes. Pakistan Journal of
Nutrition, 5(2), 166–168.
AOAC. (1984). Official Methods of Analysis 14th ed. Arlington, VA: Associ-
ation of official Analytical Chemists.
AOAC. (1995). Association of Official Analytical Chemistry (AOAC): Official
Methods of Analysis of the Association of Official Analytical Chemists
(pp. 1298).15th ed. W. Horwitz(Ed.), Arlington, VA: Association of
Official Analytical Chemists.
Bairagi, A., Ghosh, K. S., Sen, S. K., & Ray, A. K. (2002). Duckweed
(Lemna polyrhiza) leaf meal as a source of feedstuff in formulated
diets for rohu (Labeo rohita Ham.) fingerlings after fermentation with
MESHRAM ET AL.
a fish intestinal bacterium. Bioresource technology, 85(1), 17–24.
https://doi.org/10.1016/S0960-8524(02)00067-6
Beckman, B. R., & Dickhoff, W. W. (1998). Plasticity of smelting in spring
Chinook salmon: Relation to growth and insulin-like growth factor-I.
Journal of Fish Biology, 53(808), 826.
Bulow, F. J. (1987). RNA-DNA ratios as indicators of growth in fish: A
review. In R. C. Summerfelt, & G. C. Hall (Eds.), The Age and Growth
of Fish (pp. 45–64). Ames, IA: The Lowa State University Press.
Campbell, G. L., & Bedford, M. R. (1992). Enzyme applications for
monogastric feeds: A review. Canadian Journal of Animal science, 72,
449–466. https://doi.org/10.4141/cjas92-058
Clemmons, D. R., & Underwood, L. E. (1991). Nutritional regulation of
IGF-I and IGF binding proteins. Annual Review of Nutrition, 11(1),
393–412. https://doi.org/10.1146/annurev.nu.11.070191.002141
Duan, C. (1998). Nutritional and developmental regulation of insulin like
growth factors in fish. Journal of Nutrition, 128, 306S–314S. https://d
oi.org/10.1093/jn/128.2.306S
Dyer, A. R., Barlow, C. G., Bransden, M. P., Carter, C. G., Glencross, B.
D., Richardson, N., . . . Carragher, J. F. (2004). Correlation of plasma
IGF-I concentrations and growth rate in aquacultured finfish: A tool
for assessing the potential of new diets. Aquaculture, 236, 583–592.
https://doi.org/10.1016/j.aquaculture.2003.12.025
Felix, N., & Brindo, R. A. (2013). Substituting fish meal with fermented
seaweed, Kappaphycus alvarezii in diets of juvenile freshwater prawn
Macrobrachium rosenbergii. International Journal of Fisheries and
Aquatic Studies, 1, 261–265.
Gao, Y., Shang, C., Maroof, M. A., Biyashev, R. M., Grabau, E. A., Kwa-
nyuen, P., & Buss, G. R. (2007). A modified colorimetric method for
phytic acid analysis in soybean. Crop Science, 47(5), 1797–1803.
https://doi.org/10.2135/cropsci2007.03.0122
Gomez-Requeni, P., Mingarro, M., Caulduch-Giner, J. A., Me dale, F., Mar-
tin, S. A. M., Houlihan, D. F., . . . Perez-Sanchez, J. (2004). Protein
growth performance, amino acid utilisation and somatotropic axis
responsiveness to fish meal replacement by plant protein sources in
gilthead sea bream (Sparus aurata). Aquaculture, 232, 493–510.
https://doi.org/10.1016/S0044-8486(03)00532-5
Gupta, S. K., Pal, A. K., Sahu, N. P., Jha, A. K., & Kumar, S. (2015). Effects
of dietary microbial levan on growth performance, RNA/DNA ratio
and some physio-biochemical responses of Labeo rohita (Hamilton)
juveniles. Aquaculture Nutrition, 21(6), 892–903. https://doi.org/10.
1111/anu.12216
Harborne, J. B. (1973). Phenolic compounds. In Phytochemical Methods
(pp. 33–88). Dordrecht, Netherlands: Springer. https://doi.org/10.
1007/978-94-009-5921-7
Hassan, M. S., Soltan, M. A., & Abdel-Moez, A. M. (2015). Nutritive value
of soybean meal after solid state fermentation with Saccharomyces
cerevisiae for Nile tilapia, Oreochromis niloticus. Animal Feed Science
and Technology, 201, 89–98. https://doi.org/10.1016/j.anifeedsci.
2015.01.007
Hirabayashi, M., Matsui, T., & Yano, H. (1998). Fermentation of soybean
meal with Aspergillus usamii improves zinc availability in rats. Biologi-
cal trace element research, 61(2), 227–234. https://doi.org/10.1093/
ps/77.4.552
Ishida, H., Suzuno, H., Sugiyama, N., Innami, S., Tadokoro, T., & Maekawa,
A. (2000). Nutritive evaluation on chemical components of leaves stalks
and stems of sweet potatoes (Ipomoea batataspoir). Food Chemistry, 68
(3), 359–367. https://doi.org/10.1016/S0308-8146(99)00206-X
Jones, J. I., & Clemmons, D. R. (1995). Insulin-like growth factors and
their binding proteins: Biological actions. Endocrine Reviews, 16(1), 3–
34. https://doi.org/10.1210/edrv-16-1-3
Jose, S., Mohan, M. V., Shyama, S., Ramachandran Nair, K. G., & Mathew,
P. T. (2006). Effect of soybean-meal-based diets on the growth and
survival rate of the Indian major carp, Cirrhinus mrigala (Ham.). Aqua-
culture Nutrition, 12(4), 275–279. https://doi.org/10.1111/j.1365-
2095.2006.00405.x
| 9
Kiers, J. L., Meijer, J. C., Nout, M. J. R., Rombouts, F. M., Nabuurs, M. J.
A., & Van der Meulen, J. (2003). Effect of fermented soya beans on
diarrhoea and feed efficiency in weaned piglets. Journal of Applied
Microbiology, 95(3), 545–552. https://doi.org/10.1046/j.1365-2672.
2003.02011.x
Kim, B. N., Yang, J. L., & Song, Y. S. (1999). Physiological functions of
chongkukjang. Food and Nutrtion, 4, 40–46.
Kumar, V., Makkar, H. P. S., Amselgruber, W., & Becker, K. (2010). Physi-
ological, haematological and histopathological responses in common
carp (Cyprinus carpio L.) fingerlings fed with differently detoxified Jat-
ropha curcas kernel meal. Food and Chemical Toxicology, 48, 2063–
2072. https://doi.org/10.1016/j.fct.2010.05.007
Kumar, S., Sahu, N. P., Gupta, S., Deo, A. D., Shamna, N., & Ranjan, A.
(2017). Inclusion level of deoiled rice bran (DORB) in the diet of
Labeo rohita (Hamilton, 1882) fingerlings: Effect on growth and gene
expression of IGF-I and IGF-II. Aquaculture, 481, 211–217. https://d
oi.org/10.1016/j.aquaculture.2017.08.025
Kumar, S., Sahu, N. P., & Pal, A. K. (2006). Non-gelatinized Corn Supple-
mented with Microbial cc-amylase at Sub-optimal Protein in the Diet
of Labeo rohita (Hamilton) Fingerlings Increases Cell Size of Muscle.
Journal of Fisheries and Aquatic Science, 1(2), 102–111.
Kumar, S., Sahu, N. P., & Ranjan, A. (2018). Feeding de-oiled rice bran
(DORB) to Rohu, Labeo rohita: Effect of varying dietary protein and
lipid level on growth, body composition, and insulin like growth fac-
tor (IGF) expression. Aquaculture, 492, 59–66. https://doi.org/10.
1016/j.aquaculture.2018.04.001
Kumar, S., Sandor Zs, J., Nagy, Z., Fazekas, G., Havasi, M., Sinha, A. K., &
Gal, D.. (2017). Potential of processed animal protein versus soybean
meal to replace fish meal in practical diets for European catfish (Silu-
rus glanis): Growth response and liver gene expression. Aquaculture
Nutrition, 0, 1–11. https://doi.org/10.1111/anu.12487
Limbu, S. M., Shoko, A. P., Lamtane, H. A., Kishe-Machumu, M. A., Joram,
M. C., Mbonde, A. S., & Mgaya, Y. D. (2016). Supplemental effects of
mixed ingredients and rice bran on the growth performance, survival
and yield of Nile tilapia, Oreochromis niloticus reared in fertilized
earthen ponds. Springer Plus, 5(1), 5. https://doi.org/10.1186/
s40064-015-1643-x
Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene
expression data using real-time quantitative PCR and the 2 DDCT
method. Methods, 25(4), 402–408. https://doi.org/10.1006/meth.
2001.1262
Maity, J., & Patra, B. C. (2008). Effect of replacement of fishmeal by
Azolla leaf meal on growth, food utilization, pancreatic protease
activity and RNA/DNA ratio in the fingerlings of Labeo rohita (Ham.).
Canadian Journal of Pure and Applied Sciences, 2(2), 323.
Makkar, H. P., Siddhuraju, P., & Becker, K.. (2007). Plant Secondary
Metabolites (pp. 67). Louisville, KY: Humana Press
Makkar, H. P. S., & Becker, K. (2008). Protein concentrate from Jatropha
curcas screw-pressed seed cake and toxic and anti-nutritional factors
in protein concentrate. Journal of the Science of Food and Agriculture,
88, 1542–1548. https://doi.org/10.1002/jsfa.3248
Margaret, C. L., Cheng, C. H., & Chan, K. M. (2006). Effects of chronic
cysteamine treatment on growth enhancement and insulin-like
growth factor I and II mRNA levels in common carp tissues. British
Journal of Nutrition, 96(04), 650–659. https://doi.org/10.1079/
BJN20061870
Mutch, D. M., Wahli, W., & Williamson, G. (2005). Nutrigenomics and
nutrigenetics: The emerging faces of nutrition. The FASEB Journal, 19,
1602–1616. https://doi.org/10.1096/fj.05-3911rev
Mwachireya, S. A., Beames, R. M., Higgs, D. A., & Dosanjh, B. S. (1999).
Digestibility of canola protein products derived from the physical,
enzymatic and chemical processing of commercial canola meal in
rainbow trout Oncorhynchus mykiss (Walbaum) held in fresh water.
Aquaculture Nutrition, 5(2), 73–82. https://doi.org/10.1046/j.1365-
2095.1999.00089.x
10 | MESHRAM
Ndau, L. J., & Madalla, A. N. (2015). Effects of soaked pigeon peas on
the growth of Nile tilapia (Oreochromis niloticus L) fingerlings. Journal
of Fisheries & Livestock Production, 3, 125.
Ng, W. K., & Wee, K. L. (1989). The nutritive value of cassava leaf meal
in pelleted feed for Nile tilapia. Aquaculture, 83(1–2), 45–58.
https://doi.org/10.1016/0044-8486(89)90059-8
Nwosu, J. N. (2010). The effects of processing on the functional proper-
ties of ‘Oze’ (Bosqueia angolensis) seeds. Pakistan Journal of Nutrition,
9(8), 781–786. https://doi.org/10.3923/pjn.2010.781.786
Nwosu, J. N. (2011). The effects of processing on the anti-nutritional
properties of ‘Oze’ Bosqueiaangolensis seed. Journal of American
Science, 7(1), 1–6.
Olude, O. O., Alegbeleye, W. O. A., & Obasa, S. O.(2008). The use of
soaked copra meal as a partial substitute for soybean meal in the diet
of Nile tilapia (Oreochromis niloticus) fingerlings. Lipids, 8(9.45), 9–62.
Osman, M. A. (2007). Changes in nutrient composition, trypsin inhibitor,
phytate, tannins and protein digestibility of dolichos lablab seeds
(Lablab purpuresus (L) sweet) occurring during germination. Journal of
Food Technology, 5, 294–299.
Picha, M. E., Turano, M. J., Beckman, B. R., & Borski, R. J. (2008). Endo-
crine biomarkers of growth and applications to aquaculture: A mini
review of growth hormone, insulin-like growth factor (IGF)-I, and
IGF-binding proteins as potential growth indicators in fish. North
American Journal of Aquaculture, 70(2), 196–211. https://doi.org/10.
1577/A07-038.1
Pierce, A. L., Breves, J. P., Moriyama, S., Hirano, T., & Grau, E. G. (2011).
Differential regulation of Igf1 and Igf2 mRNA levels in tilapia hepato-
cytes: Effects of insulin and cortisol on GH sensitivity. Journal of
Endocrinology, 211(2), 201–210. https://doi.org/10.1530/JOE-10-
0456
Ramachandran, S., Bairagi, A., & Ray, A. K. (2005). Improvement of nutri-
tive value of grass pea (Lathyrus sativus) seed meal in the formulated
diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation
with a fish gut bacterium. Bioresource Technology, 96, 1465–1472.
https://doi.org/10.1016/j.biortech.2004.12.002
Ramakrishna, R., Shipton, T. A., & Hasan, M. R.. (2013). Feeding and feed
management of Indian major carps in Andhra Pradesh, India. Food
and Agriculture Organization of the United Nations. FAO Fisheries
and Aquaculture Technical Paper No. 578. Rome, FAO, p. 90.
Schneider, W. C.. (1957). Determination of nucleic acids in tissue by pan-
tose analysis. In Calowick, S. P., & Kaplon, N. O. (Eds.), Methods of
Enzymology (pp. 680). New York, NY: Academic Press.
Shamna, N., Sardar, P., Sahu, N. P., Pal, A. K., Jain, K. K., & Phulia, V.
(2015). Nutritional evaluation of fermented Jatropha protein concen-
trates in Labeo rohita fingerlings. Aquaculture Nutrition, 21(1), 33–42.
https://doi.org/10.1111/anu.12138
Singh, K., Garg, S. K., Bhatnagar, A., & Kalla, A. (2004). Comparison of
five different practical diets with various concentrations of dietary
protein in nursery ponds: Survival and growth of Indian major carp
fry. Asian Fisheries Science, 17(1/2), 121–134.
SOFIA. (2016). The State of World Fisheries and Aquaculture. Contributing
to Food Security and Nutrition for All (pp. 20). Rome, Italy: FAO.
Sun, H., Tang, J. W., Yao, X. H., W. U., Y. F., Wang, X., Liu, Y., & Lou, B..
(2015). Partial substitution of fish meal with fermented cottonseed
meal in juvenile black sea bream (Acanthopagrus schlegelii) diets,
Aquaculture, 446, 30–36. https://doi.org/10.1016/j.aquaculture.2015.
04.020
ET AL.
Tanaka, Y., Gwak, W. S., Tanaka, M., Sawada, Y., Okada, T., Miyashita, S.,
& Kumai, H. (2007). Ontogenetic changes in RNA, DNA and protein
contents of laboratory-reared Pacific bluefin tuna Thunnus orientalis.
Fisheries Science, 73(2), 378–384. https://doi.org/10.1111/j.1444-
2906.2007.01345.x
Tatar, M., Bartke, A., & Antebi, A. (2003). The endocrine regulation of
aging by insulin-like signals. Science, 299, 1346–1351. https://doi.
org/10.1126/science.108144
Uchida, K., Kajimura, S., Riley, L. G., Hirano, T., Aida, K., & Grau, E. G.
(2003). Effects of fasting on growth hormone/insulin-like growth
factor I axis in the tilapia, Oreochromis mossambicus. Comparative
Biochemistry and Physiology Part A: Molecular & Integrative Physiol-
ogy, 134(2), 429–439. https://doi.org/10.1016/S1095-6433(02)
00318-5
Veerina, S. S., Nandeesha, M. C., & Gopal Rao, K.. (1993). Status and
Technology of Indian Major Carp Farming in Andhra Pradesh, India
(pp.52). Special Publication No. 9. Andhra Pradesh, India: Asian Fish-
eries Society Special Publication.
Vhanalakar, S. A., & Muley, D. V. (2014). Effect of Dietary Incorporation
of Gliricidia Maculata Leaf Meal on Growth and Feed Utilization of
Cirrhinus Mrigala Fingerlings. Global Journal of Science Frontier
Research Biological Science, 14(1), 2014.
Vijayakumari, K., Siddhuraju, P., Pugalenthi, M., & Janardhanan, K..
(1998). Effect of soaking and heat processing on the levels of antinu-
trients and digestible proteins in seeds of Vigna aconitifolia and Vigna
sinensis. Food Chemistry, 63(2), 259–264. https://doi.org/10.1016/
S0308-8146(97)00207-0
Wang, J. H., Guo, H., Zhang, T. R., Wang, H., Liu, B. N., & Xiao, S..
(2017). Growth performance and digestion improvement of juvenile
sea cucumber Apostichopus japonicus fed by solid-state fermentation
diet. Aquaculture Nutrition, 23, 1312–1318. https://doi.org/10.1111/
anu.12506
Yamamoto, T., Iwashita, Y., Matsunari, H., Sugita, T., Furuita, H., Akimoto,
A., & Suzuki, N. (2010). Influence of fermentation conditions for soy-
bean meal in a non-fish meal diet on the growth performance and
physiological condition of rainbow trout Oncorhynchus mykiss. Aqua-
culture, 309(1), 173–180. https://doi.org/10.1016/j.aquaculture.2010.
09.021
Yuan, Y. C., Lin, Y. C., Yang, H. J., Gong, Y., Gong, S. Y., & Yu, D. H.
(2013). Evaluation of fermented soybean meal in the practical diets
for juvenile Chinese sucker, Myxocyprinus asiaticus. Aquaculture
Nutrition, 19(1), 74–83. https://doi.org/10.1111/j.1365-2095.2012.
00939.x
How to cite this article: Meshram S, Deo AD, Kumar S,
Aklakur M, Sahu NP. Replacement of de-oiled rice bran by
soaked and fermented sweet potato leaf meal: Effect on
growth performance, body composition and expression of
insulin-like growth factor 1 in Labeo rohita (Hamilton),
fingerlings. Aquac Res. 2018;00:1–10.
https://doi.org/10.1111/are.13735