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 Abstract
Education, Versova, Mumbai, India A 60-day feeding trial was conducted to determine the growth performance and
Correspondence expression of insulin-like growth factor 1 gene (IGF-I gene) in Labeo rohita fingerlings
Ashutosh D. Deo, Fish Nutrition, fed with either raw, soaked or fermented sweet potato leaf meal (SPLM) by com-
Biochemistry and Physiology Division, ICAR-
Central Institute of Fisheries Education, pletely replacing de-oiled rice bran (DORB), following a completely randomized
Versova, Mumbai, 400061, India. design. Seven isonitrogenous (30%) and isocaloric (1.8 MJ/100 g) diets were pre-
E-mails: [email protected];
[email protected] 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 %,
Funding information
Indian Council of Agricultural Research SGR (specific growth rate) and PER (protein efficiency ratio) were significantly
(ICAR) (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 important cultivable species and high consumer demand due to its
taste preference. The aquaculture has also grown rapidly and led to
Fish farming is the fastest growing agricultural sector in the world more feed based culture system. Feeding is the most important fac-
(SOFIA, 2016). The increasing human population has a lead increased tors required for faster growth and higher yield of cultured Labeo
demand for quality protein. Fish being a good protein source has rohita (Jose, Mohan, Shyama, Ramachandran Nair, & Mathew, 2006;
witnessed an increase in per capita apparent fish consumption from Maity & Patra, 2008). Aquafeed ingredients mainly depend on oil
an average of 9.9 kg in the 1960s to 20 kg in 2016 (SOFIA, 2016). cake as the main conventional protein sources and de-oiled rice bran
India is dominating in aquaculture production, which includes many (DORB) as an energy source for the carp culture in the Asian sub-
species like carps, catfishes, shellfish, pangasius, tilapia etc. Among continent. As a practice, DORB (De-Oiled Rice Bran) is most com-
the three Indian major carps (IMC), Labeo rohita is the most 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 | MESHRAM ET AL.

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

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

2.3 | Diet preparation 2.4 | Proximate composition and antinutritional

factor
Seven isonitrogenous (30% CP) and isocaloric (1.8 MJ/100 g) practi-
cal diets were prepared (Table 1) with the replacement of de-oiled Proximate composition of the experimental diets and fish were anal-
rice bran with raw (R), soaked (S) and fermented (F) SPLM viz. C ysed following AOAC (1995). The experimental diets and body sam-
(control with DORB), TR50 (treatment with 50% raw SPLM in 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. | 5

(1957) and was calculated as follows: The DNA content of the

3 | RESULTS
nucleic acid extract is presented by the following equation:

lgDNA mL1 ¼ 1=4 OD at 600nm  0:019 3.1 | Proximate composition of diets and fish body

The RNA content of the nucleic acid extract is presented by the Proximate composition of diets and SPLM, DORB are presented in
following equation: 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-
lgRNA mL1 ¼ ½ðOD at 660nm þ 0:0081Þ tively but the crude fibre decreased from 8.30% to 6.01% (Table 3).
 ðlgDNA mL1  0:013Þ=0:116 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
2.8 | RNA extraction and cDNA synthesis
from 8.30% to 8.10% (Table 3). The whole body compositions of all
Approximately 100 mg of liver tissue was mechanically homogenized the experimental fish of different treatments are presented in
with 1 ml of the TRI reagent (Sigma, St. Louis, MO, USA) and the Table 4. Dietary replacement of de-oiled rice bran by SPLM did not
total RNA was extracted according to the manufacturer’s instruc- affect (p > 0.05) the moisture, crude protein and lipid. However, the
tions. The purity and concentration of RNA were checked in Nano- ash content was significantly higher in TF100 which is similar to
Drop spectrophotometer (Thermoscientific, USA). The RNA samples TF50. The crude protein content varied from 14.8% to 16.4%,
were treated with DNase (Fermentas, USA) according to the proto- whereas lipid content of all the fish was estimated within 3.64% to
col for the removal of residual DNA during preparation. cDNA was 4.3% and total ash content varied from 3.2% to 4.88%. The reduc-
synthesized from the 1 lg of total RNA using oligo dT primer follow- tion in antinutritional factor (phytate, oxalate, alkaloid, tannin and
ing the protocol of first stand cDNA synthesis kit (Fermentas, USA). HCN) after soaking and fermentation are presented in Table 3.

2.9 | Quantitative RT-PCR 3.2 | Growth performance

The relative quantification of IGF-I mRNA expression was performed There was significant difference (p < 0.05) in the body weight gain
using AriaMx Real-Time PCR System (Agilent Technology, LDA among the different treatment groups at the end of the experiment
Malaysia) using SYBER Green qPCR master mix without ROX (Ther- (Table 5). The weight gain % specific growth rate and PER in TF100
moscientific, USA). Standard reaction mixtures (10 µl) were assem- were significantly (p < 0.05) higher as compared to other treatment
bled using 5 µl of SYBER Green supermix 29, 1 µl each primer groups. However, the highest FCR was recorded in TR100 group,
(5 pm) and 100 ng of template cDNA For amplification of IGF-I which was similar to TR50. The lower FCR was recorded in TF50-
gene-specific primer and beta-actin reference gene primer (Table 2) and TF100-fed group which is statically similar to TS50- and TS100-
were designed using Gene runner software (version 3.05) based on fed group.
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- 3.3 | Body indices
parative CT values were used to estimate the relative expression of The HSI value of different treatment was not significantly different
mRNA. All reaction run in duplicates and the data were analysed (p > 0.05) (Table 5) among the treatment groups but the ISI value of
using the 2DDct method (Livak & Schmittgen, 2001). different experimental groups varied significantly (p < 0.05). In this
study, the ISI value in TF50 fed group was significantly (p < 0.05)

2.10 | Statistical analysis higher and lowest value was recorded in control-fed group.

All values presented are the mean
standard error. Statistical analy-
sis of data was performed by analysis of variance (ANOVA) in SPSS 3.4 | RNA, DNA and RNA/DNA ratio
(Version 22) followed by Duncan’s multiple range test. Significance The RNA and RNA/DNA ratio varied significantly (p < 0.05) in the
was considered at p < 0.05. different treatment groups (Table 6). The DNA content did not show

T A B L E 2 Primer used for

Gene Accession no. Primer sequence Amplicon size Anneline temp.
expression study in real time PCR
IGF-I KX455870 F-GCAAACCGACAGGCTATGGGC 166 57
R-GTGTCTGTGTGCCGTTCCGC
b-actin EU184877 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. growth performance of animal. Therefore, to increase SPLM nutri-
However, the RNA concentration and RNA/DNA ratio was found tional value two method, microbial fermentation and water soaking
significantly higher in the treated SPLM fed groups compared to the was planned. It was for replacing the expensive protein resources by
control group and raw SPLM-fed groups. The RNA concentration SPLM to make cost effective feed for animal feed production. Reduc-
was found significantly higher in TS50-fed group followed by TS100 tion in antinutritional factors (phytate, tannin, alkaloid, HCN and oxa-
treatment groups. RNA/DNA ratio was observed significantly higher late) from plant ingredients for sustainability have been taken by many
in TF100-fed groups. 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
3.5 | Expression of IGF-I mRNA of Labeo rohita
factors. Similar to earlier studies (Vijayakumari, Siddhuraju, Pugalenthi,
liver tissue
& Janardhanan, 1998; Nwosu, 2010), soaking increased the nutritional
The expression of IGF-I in liver tissue of L. rohita varied significantly value of SPLM by decreasing the antinutritional factors (Table 3).
(p < 0.05) among the treatment groups (Figure 1). IGF-I mRNA Solid-state fermentation is also important and an effective method for
expression was significantly (p < 0.05) higher in TF100 and TS100, reducing antinutritional factors to improve nutritional value (Yuan
while control, TR50- and TR100-fed groups showed lower value et al., 2013). In the present study, crude protein and lipid content in
(Figure 1). 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-
4 | DISCUSSION ment. Previous studies focused on the replacement of ingredients in
the feed, based on the growth data of the experimental animal. Soak-
SPLM is important source of protein and energy for fish feed but some ing and solid-state fermentation (SSF) are well-documented for the
antinutritional factor reduced the nutritional value and inhibited 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 | MESHRAM ET AL.

Olude, Alegbeleye and Obasa (2008) found soaked copra meal can 5 | CONCLUSION
be incorporated in the diet of Nile tilapia up to 30% without any
deleterious effect on growth performance and nutrient utilization. It The present study clearly established that the SPLM has potential as
is important to mention that in the present study, the DORB was aquafeed ingredients. The fermented sweet potato leaf meal using
replaced with raw and processed sweet potato leaf meal which is Chaetomium globosum can replace 100% de-oiled rice bran in the
first attempts of this kind hence; in the absence of authentic pub- diet of L. rohita without any detrimental effect on growth perfor-
lished literature, it could not be compared or discussed in particular mance, nutrient utilization which was also verified with the expres-
to carp feed. sion of growth gene IGF-I. However, further study needs to be
Several studies considered RNA/DNA ratio as an index of conducted on economic feasibility of SPLM as a replacer of DORB
growth (Akhtar, Pal, Sahu, Ciji, & Gupta, 2012; Gupta, Pal, Sahu, Jha, and also to evaluate the long-term feeding effect of fermented
& Kumar, 2015; Kumar, Sahu, & Ranjan, 2018; Kumar, Sandor Zs, sweet potato leaf meal on the growth and other related parameters
et al., 2017; Kumar, Sahu, et al., 2017). The quantitative analysis of of fish before SPLM is recommended for commercial application in
RNA and DNA can disclose the growth rate in fish as RNA is an the carp diets.
organizer of protein synthesis. The somatic growths increased with
increased RNA content (Bulow, 1987) while the DNA content
ACKNOWLEDGMENTS
remains constant. RNA is organizer molecule for protein synthesis.
The RNA/DNA ratio is a sensitive index of anabolic activity and The Authors are thankful to Indian Council of Agricultural Research
highly varies depending on the nutritional status and feeding (Tanaka (ICAR) for the financial support. Director of ICAR- CIFE (Indian
et al., 2007). The present study indicated that the RNA concentra- Council of Agricultural Research-Central Institute of Fisheries Educa-
tion and RNA/DNA ratio were significantly higher in soaked and tion) for providing the necessary infrastructural facilities for complet-
solid state fermented SPLM replacing DORB-fed group as compared ing the experiment.
to control and raw. Similarly, the RNA/DNA ratio indicates the
growth performance in fish L. rohita (Akhtar et al., 2012; Kumar,
CONFLICT OF INTEREST
Sahu, & Pal, 2006; Kumar, Sandor Zs, et al., 2017; Kumar, Sahu,
et al., 2017). The authors declare no competing financial interests.
As the growth of animal is affected by many factors, evaluation
of growth only by considering weight gain of the animal due to feed-
ORCID
ing of particular ingredients may not be justified. Hence, molecular
responses of the growth-regulated gene are being used to choose Ashutosh D Deo http://orcid.org/0000-0002-9341-2084
for evaluation of particular ingredients in the feed of a targeted spe- Narottam P Sahu http://orcid.org/0000-0003-3932-7302
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 REFERENCES
factors (IGF), IGF-I belong to the growth regulating family. These
Adewolu, M. A. (2008). Potentials of sweet potato (Ipomoea batatas) leaf
hormones are synthesized in liver and its expression is controlled by meal as dietary ingredient for Tilapia zilli fingerlings. Pakistan Journal
the quantity of feed fed and its nutrient composition in fish and of Nutrition, 7(3), 444–449. https://doi.org/10.3923/pjn.2008.444.
birds (Clemmons & Underwood, 1991). The expression of IGF-I con- 449
Akhtar, M. S., Pal, A. K., Sahu, N. P., Ciji, A., & Gupta, S. K. (2012). Effects
centration was positively correlated with the growth rate in Salmo
of dietary pyridoxine on growth and biochemical responses of Labeo
salar (Dyer et al., 2004). Dietary protein level alters the expression rohita fingerlings exposed to endosulfan. Pesticide biochemistry and
of IGF-I in fish (Gomez-Requeni et al., 2004) and also interacts with physiology, 103(1), 23–30. https://doi.org/10.1016/j.pestbp.2012.02.
nutritional status of fish (Margaret, Cheng, & Chan, 2006). Insulin- 004
Almazan, A. M. (1995). Antinutritional factors in sweet potato greens.
like growth factor I axis is regulated by the nutritional and metabolic
Journal of Food Composition and Analysis, 8(4), 363–368. https://doi.
condition. Its subsequent effect on liver and liver for modulating org/10.1006/jfca.1995.1031
sensitivity to growth hormone is a real concern (Pierce, Breves, Mor- Antia, B. S., Akpanz, E. J., Okonl, P. A., & Umorenl, I. U. (2006). Nutritive
iyama, Hirano, & Grau, 2011). In the present study highest expres- and Anti-Nutritive Evaluation of Sweet Potatoes. Pakistan Journal of
Nutrition, 5(2), 166–168.
sion (p < 0.05) of IGF-I mRNA was observed in TF100-fed group.
AOAC. (1984). Official Methods of Analysis 14th ed. Arlington, VA: Associ-
Similar observation has been made by Kumar, Sandor Zs, et al., ation of official Analytical Chemists.
2017; Kumar, Sahu, et al., 2017 who reported decreased growth AOAC. (1995). Association of Official Analytical Chemistry (AOAC): Official
performance with decreased IGF-I gene expression in Silurus glanis Methods of Analysis of the Association of Official Analytical Chemists
(pp. 1298).15th ed. W. Horwitz(Ed.), Arlington, VA: Association of
when fishmeal was replaced with soyabean meal. The IGF-I mRNA
Official Analytical Chemists.
expression is an important indicator of growth in Tilapia (Uchida
Bairagi, A., Ghosh, K. S., Sen, S. K., & Ray, A. K. (2002). Duckweed
et al., 2003); Labeo rohita (Kumar, Sandor Zs, et al., 2017; Kumar, (Lemna polyrhiza) leaf meal as a source of feedstuff in formulated
Sahu, et al., 2017). diets for rohu (Labeo rohita Ham.) fingerlings after fermentation with
MESHRAM ET AL. | 9

a fish intestinal bacterium. Bioresource technology, 85(1), 17–24. Kiers, J. L., Meijer, J. C., Nout, M. J. R., Rombouts, F. M., Nabuurs, M. J.
https://doi.org/10.1016/S0960-8524(02)00067-6 A., & Van der Meulen, J. (2003). Effect of fermented soya beans on
Beckman, B. R., & Dickhoff, W. W. (1998). Plasticity of smelting in spring diarrhoea and feed efficiency in weaned piglets. Journal of Applied
Chinook salmon: Relation to growth and insulin-like growth factor-I. Microbiology, 95(3), 545–552. https://doi.org/10.1046/j.1365-2672.
Journal of Fish Biology, 53(808), 826. 2003.02011.x
Bulow, F. J. (1987). RNA-DNA ratios as indicators of growth in fish: A Kim, B. N., Yang, J. L., & Song, Y. S. (1999). Physiological functions of
review. In R. C. Summerfelt, & G. C. Hall (Eds.), The Age and Growth chongkukjang. Food and Nutrtion, 4, 40–46.
of Fish (pp. 45–64). Ames, IA: The Lowa State University Press. Kumar, V., Makkar, H. P. S., Amselgruber, W., & Becker, K. (2010). Physi-
Campbell, G. L., & Bedford, M. R. (1992). Enzyme applications for ological, haematological and histopathological responses in common
monogastric feeds: A review. Canadian Journal of Animal science, 72, carp (Cyprinus carpio L.) fingerlings fed with differently detoxified Jat-
449–466. https://doi.org/10.4141/cjas92-058 ropha curcas kernel meal. Food and Chemical Toxicology, 48, 2063–
Clemmons, D. R., & Underwood, L. E. (1991). Nutritional regulation of 2072. https://doi.org/10.1016/j.fct.2010.05.007
IGF-I and IGF binding proteins. Annual Review of Nutrition, 11(1), Kumar, S., Sahu, N. P., Gupta, S., Deo, A. D., Shamna, N., & Ranjan, A.
393–412. https://doi.org/10.1146/annurev.nu.11.070191.002141 (2017). Inclusion level of deoiled rice bran (DORB) in the diet of
Duan, C. (1998). Nutritional and developmental regulation of insulin like Labeo rohita (Hamilton, 1882) fingerlings: Effect on growth and gene
growth factors in fish. Journal of Nutrition, 128, 306S–314S. https://d expression of IGF-I and IGF-II. Aquaculture, 481, 211–217. https://d
oi.org/10.1093/jn/128.2.306S oi.org/10.1016/j.aquaculture.2017.08.025
Dyer, A. R., Barlow, C. G., Bransden, M. P., Carter, C. G., Glencross, B. Kumar, S., Sahu, N. P., & Pal, A. K. (2006). Non-gelatinized Corn Supple-
D., Richardson, N., . . . Carragher, J. F. (2004). Correlation of plasma mented with Microbial cc-amylase at Sub-optimal Protein in the Diet
IGF-I concentrations and growth rate in aquacultured finfish: A tool of Labeo rohita (Hamilton) Fingerlings Increases Cell Size of Muscle.
for assessing the potential of new diets. Aquaculture, 236, 583–592. Journal of Fisheries and Aquatic Science, 1(2), 102–111.
https://doi.org/10.1016/j.aquaculture.2003.12.025 Kumar, S., Sahu, N. P., & Ranjan, A. (2018). Feeding de-oiled rice bran
Felix, N., & Brindo, R. A. (2013). Substituting fish meal with fermented (DORB) to Rohu, Labeo rohita: Effect of varying dietary protein and
seaweed, Kappaphycus alvarezii in diets of juvenile freshwater prawn lipid level on growth, body composition, and insulin like growth fac-
Macrobrachium rosenbergii. International Journal of Fisheries and tor (IGF) expression. Aquaculture, 492, 59–66. https://doi.org/10.
Aquatic Studies, 1, 261–265. 1016/j.aquaculture.2018.04.001
Gao, Y., Shang, C., Maroof, M. A., Biyashev, R. M., Grabau, E. A., Kwa- Kumar, S., Sandor Zs, J., Nagy, Z., Fazekas, G., Havasi, M., Sinha, A. K., &
nyuen, P., & Buss, G. R. (2007). A modified colorimetric method for Gal, D.. (2017). Potential of processed animal protein versus soybean
phytic acid analysis in soybean. Crop Science, 47(5), 1797–1803. meal to replace fish meal in practical diets for European catfish (Silu-
https://doi.org/10.2135/cropsci2007.03.0122 rus glanis): Growth response and liver gene expression. Aquaculture
Gomez-Requeni, P., Mingarro, M., Caulduch-Giner, J. A., Me dale, F., Mar- Nutrition, 0, 1–11. https://doi.org/10.1111/anu.12487
tin, S. A. M., Houlihan, D. F., . . . Perez-Sanchez, J. (2004). Protein Limbu, S. M., Shoko, A. P., Lamtane, H. A., Kishe-Machumu, M. A., Joram,
growth performance, amino acid utilisation and somatotropic axis M. C., Mbonde, A. S., & Mgaya, Y. D. (2016). Supplemental effects of
responsiveness to fish meal replacement by plant protein sources in mixed ingredients and rice bran on the growth performance, survival
gilthead sea bream (Sparus aurata). Aquaculture, 232, 493–510. and yield of Nile tilapia, Oreochromis niloticus reared in fertilized
https://doi.org/10.1016/S0044-8486(03)00532-5 earthen ponds. Springer Plus, 5(1), 5. https://doi.org/10.1186/
Gupta, S. K., Pal, A. K., Sahu, N. P., Jha, A. K., & Kumar, S. (2015). Effects s40064-015-1643-x
of dietary microbial levan on growth performance, RNA/DNA ratio Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene
and some physio-biochemical responses of Labeo rohita (Hamilton) expression data using real-time quantitative PCR and the 2 DDCT
juveniles. Aquaculture Nutrition, 21(6), 892–903. https://doi.org/10. method. Methods, 25(4), 402–408. https://doi.org/10.1006/meth.
1111/anu.12216 2001.1262
Harborne, J. B. (1973). Phenolic compounds. In Phytochemical Methods Maity, J., & Patra, B. C. (2008). Effect of replacement of fishmeal by
(pp. 33–88). Dordrecht, Netherlands: Springer. https://doi.org/10. Azolla leaf meal on growth, food utilization, pancreatic protease
1007/978-94-009-5921-7 activity and RNA/DNA ratio in the fingerlings of Labeo rohita (Ham.).
Hassan, M. S., Soltan, M. A., & Abdel-Moez, A. M. (2015). Nutritive value Canadian Journal of Pure and Applied Sciences, 2(2), 323.
of soybean meal after solid state fermentation with Saccharomyces Makkar, H. P., Siddhuraju, P., & Becker, K.. (2007). Plant Secondary
cerevisiae for Nile tilapia, Oreochromis niloticus. Animal Feed Science Metabolites (pp. 67). Louisville, KY: Humana Press
and Technology, 201, 89–98. https://doi.org/10.1016/j.anifeedsci. Makkar, H. P. S., & Becker, K. (2008). Protein concentrate from Jatropha
2015.01.007 curcas screw-pressed seed cake and toxic and anti-nutritional factors
Hirabayashi, M., Matsui, T., & Yano, H. (1998). Fermentation of soybean in protein concentrate. Journal of the Science of Food and Agriculture,
meal with Aspergillus usamii improves zinc availability in rats. Biologi- 88, 1542–1548. https://doi.org/10.1002/jsfa.3248
cal trace element research, 61(2), 227–234. https://doi.org/10.1093/ Margaret, C. L., Cheng, C. H., & Chan, K. M. (2006). Effects of chronic
ps/77.4.552 cysteamine treatment on growth enhancement and insulin-like
Ishida, H., Suzuno, H., Sugiyama, N., Innami, S., Tadokoro, T., & Maekawa, growth factor I and II mRNA levels in common carp tissues. British
A. (2000). Nutritive evaluation on chemical components of leaves stalks Journal of Nutrition, 96(04), 650–659. https://doi.org/10.1079/
and stems of sweet potatoes (Ipomoea batataspoir). Food Chemistry, 68 BJN20061870
(3), 359–367. https://doi.org/10.1016/S0308-8146(99)00206-X Mutch, D. M., Wahli, W., & Williamson, G. (2005). Nutrigenomics and
Jones, J. I., & Clemmons, D. R. (1995). Insulin-like growth factors and nutrigenetics: The emerging faces of nutrition. The FASEB Journal, 19,
their binding proteins: Biological actions. Endocrine Reviews, 16(1), 3– 1602–1616. https://doi.org/10.1096/fj.05-3911rev
34. https://doi.org/10.1210/edrv-16-1-3 Mwachireya, S. A., Beames, R. M., Higgs, D. A., & Dosanjh, B. S. (1999).
Jose, S., Mohan, M. V., Shyama, S., Ramachandran Nair, K. G., & Mathew, Digestibility of canola protein products derived from the physical,
P. T. (2006). Effect of soybean-meal-based diets on the growth and enzymatic and chemical processing of commercial canola meal in
survival rate of the Indian major carp, Cirrhinus mrigala (Ham.). Aqua- rainbow trout Oncorhynchus mykiss (Walbaum) held in fresh water.
culture Nutrition, 12(4), 275–279. https://doi.org/10.1111/j.1365- Aquaculture Nutrition, 5(2), 73–82. https://doi.org/10.1046/j.1365-
2095.2006.00405.x 2095.1999.00089.x
10 | MESHRAM ET AL.

Ndau, L. J., & Madalla, A. N. (2015). Effects of soaked pigeon peas on Tanaka, Y., Gwak, W. S., Tanaka, M., Sawada, Y., Okada, T., Miyashita, S.,
the growth of Nile tilapia (Oreochromis niloticus L) fingerlings. Journal & Kumai, H. (2007). Ontogenetic changes in RNA, DNA and protein
of Fisheries & Livestock Production, 3, 125. contents of laboratory-reared Pacific bluefin tuna Thunnus orientalis.
Ng, W. K., & Wee, K. L. (1989). The nutritive value of cassava leaf meal Fisheries Science, 73(2), 378–384. https://doi.org/10.1111/j.1444-
in pelleted feed for Nile tilapia. Aquaculture, 83(1–2), 45–58. 2906.2007.01345.x
https://doi.org/10.1016/0044-8486(89)90059-8 Tatar, M., Bartke, A., & Antebi, A. (2003). The endocrine regulation of
Nwosu, J. N. (2010). The effects of processing on the functional proper- aging by insulin-like signals. Science, 299, 1346–1351. https://doi.
ties of ‘Oze’ (Bosqueia angolensis) seeds. Pakistan Journal of Nutrition, org/10.1126/science.108144
9(8), 781–786. https://doi.org/10.3923/pjn.2010.781.786 Uchida, K., Kajimura, S., Riley, L. G., Hirano, T., Aida, K., & Grau, E. G.
Nwosu, J. N. (2011). The effects of processing on the anti-nutritional (2003). Effects of fasting on growth hormone/insulin-like growth
properties of ‘Oze’ Bosqueiaangolensis seed. Journal of American factor I axis in the tilapia, Oreochromis mossambicus. Comparative
Science, 7(1), 1–6. Biochemistry and Physiology Part A: Molecular & Integrative Physiol-
Olude, O. O., Alegbeleye, W. O. A., & Obasa, S. O.(2008). The use of ogy, 134(2), 429–439. https://doi.org/10.1016/S1095-6433(02)
soaked copra meal as a partial substitute for soybean meal in the diet 00318-5
of Nile tilapia (Oreochromis niloticus) fingerlings. Lipids, 8(9.45), 9–62. Veerina, S. S., Nandeesha, M. C., & Gopal Rao, K.. (1993). Status and
Osman, M. A. (2007). Changes in nutrient composition, trypsin inhibitor, Technology of Indian Major Carp Farming in Andhra Pradesh, India
phytate, tannins and protein digestibility of dolichos lablab seeds (pp.52). Special Publication No. 9. Andhra Pradesh, India: Asian Fish-
(Lablab purpuresus (L) sweet) occurring during germination. Journal of eries Society Special Publication.
Food Technology, 5, 294–299. Vhanalakar, S. A., & Muley, D. V. (2014). Effect of Dietary Incorporation
Picha, M. E., Turano, M. J., Beckman, B. R., & Borski, R. J. (2008). Endo- of Gliricidia Maculata Leaf Meal on Growth and Feed Utilization of
crine biomarkers of growth and applications to aquaculture: A mini Cirrhinus Mrigala Fingerlings. Global Journal of Science Frontier
review of growth hormone, insulin-like growth factor (IGF)-I, and Research Biological Science, 14(1), 2014.
IGF-binding proteins as potential growth indicators in fish. North Vijayakumari, K., Siddhuraju, P., Pugalenthi, M., & Janardhanan, K..
American Journal of Aquaculture, 70(2), 196–211. https://doi.org/10. (1998). Effect of soaking and heat processing on the levels of antinu-
1577/A07-038.1 trients and digestible proteins in seeds of Vigna aconitifolia and Vigna
Pierce, A. L., Breves, J. P., Moriyama, S., Hirano, T., & Grau, E. G. (2011). sinensis. Food Chemistry, 63(2), 259–264. https://doi.org/10.1016/
Differential regulation of Igf1 and Igf2 mRNA levels in tilapia hepato- S0308-8146(97)00207-0
cytes: Effects of insulin and cortisol on GH sensitivity. Journal of Wang, J. H., Guo, H., Zhang, T. R., Wang, H., Liu, B. N., & Xiao, S..
Endocrinology, 211(2), 201–210. https://doi.org/10.1530/JOE-10- (2017). Growth performance and digestion improvement of juvenile
0456 sea cucumber Apostichopus japonicus fed by solid-state fermentation
Ramachandran, S., Bairagi, A., & Ray, A. K. (2005). Improvement of nutri- diet. Aquaculture Nutrition, 23, 1312–1318. https://doi.org/10.1111/
tive value of grass pea (Lathyrus sativus) seed meal in the formulated anu.12506
diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation Yamamoto, T., Iwashita, Y., Matsunari, H., Sugita, T., Furuita, H., Akimoto,
with a fish gut bacterium. Bioresource Technology, 96, 1465–1472. A., & Suzuki, N. (2010). Influence of fermentation conditions for soy-
https://doi.org/10.1016/j.biortech.2004.12.002 bean meal in a non-fish meal diet on the growth performance and
Ramakrishna, R., Shipton, T. A., & Hasan, M. R.. (2013). Feeding and feed physiological condition of rainbow trout Oncorhynchus mykiss. Aqua-
management of Indian major carps in Andhra Pradesh, India. Food culture, 309(1), 173–180. https://doi.org/10.1016/j.aquaculture.2010.
and Agriculture Organization of the United Nations. FAO Fisheries 09.021
and Aquaculture Technical Paper No. 578. Rome, FAO, p. 90. Yuan, Y. C., Lin, Y. C., Yang, H. J., Gong, Y., Gong, S. Y., & Yu, D. H.
Schneider, W. C.. (1957). Determination of nucleic acids in tissue by pan- (2013). Evaluation of fermented soybean meal in the practical diets
tose analysis. In Calowick, S. P., & Kaplon, N. O. (Eds.), Methods of for juvenile Chinese sucker, Myxocyprinus asiaticus. Aquaculture
Enzymology (pp. 680). New York, NY: Academic Press. Nutrition, 19(1), 74–83. https://doi.org/10.1111/j.1365-2095.2012.
Shamna, N., Sardar, P., Sahu, N. P., Pal, A. K., Jain, K. K., & Phulia, V. 00939.x
(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 How to cite this article: Meshram S, Deo AD, Kumar S,
five different practical diets with various concentrations of dietary Aklakur M, Sahu NP. Replacement of de-oiled rice bran by
protein in nursery ponds: Survival and growth of Indian major carp
soaked and fermented sweet potato leaf meal: Effect on
fry. Asian Fisheries Science, 17(1/2), 121–134.
SOFIA. (2016). The State of World Fisheries and Aquaculture. Contributing growth performance, body composition and expression of
to Food Security and Nutrition for All (pp. 20). Rome, Italy: FAO. insulin-like growth factor 1 in Labeo rohita (Hamilton),
Sun, H., Tang, J. W., Yao, X. H., W. U., Y. F., Wang, X., Liu, Y., & Lou, B.. fingerlings. Aquac Res. 2018;00:1–10.
(2015). Partial substitution of fish meal with fermented cottonseed
https://doi.org/10.1111/are.13735
meal in juvenile black sea bream (Acanthopagrus schlegelii) diets,
Aquaculture, 446, 30–36. https://doi.org/10.1016/j.aquaculture.2015.
04.020