Introduction
Aquaculture contributes significantly to poverty reduction and
nutritional security, hence increasing impoverished people's socioeconomic
condition, particularly in developing nations (FAO 2018). In 2018, global fish
production reached 179 MMT, with 82 MMT produced from the aquaculture industry
(FAO 2020). Increased feed-based aquaculture practices allowed for this
expansion. Aquafeeds must combine unproven unconventional locally available
components because a source of protein and energy typical dietary items are
currently in short supply. Traditional feedstuffs for carp production in Asia
include oil cakes and deoiled rice bran (DORB), which
is mostly utilized in aquafeeds (Meshram et al.
2018). Farm-made carp feed, on the other hand, typically contains more than 85
percent DORB. As a result, fish feed made in farms is highly dependent on DORB
may pose challenges about availability in the future. The use of DORB in human
food and the terrestrial livestock feed industry may also result in decreased
accessibility to DORB as an aquafeed ingredient (Maiti
et al. 2019). Thus, alternatives to DORB have to be identified. The
unused and wasted plant leaves in the form of dried meal may be a suitable
option for DORB replacement in the fish diet (Meshram
et al. 2018; Ahmad et al. 2019; Maiti et
al. 2019; Sahoo et al. 2020; Anand et al. 2020).
Due to the existence of antinutritional influences, plant leaves have
limitations for optimal inclusion in aquafeed (ANFs). Sweet potato, Leucaena
leucocephala, Morus esculenta,
Medicago sativa, Moringa oleifera, and Manihot esculenta
are just a few of the plant leaves that have lately been added into fish diets
(Ali et al. 2003; Bairagi et al. 2004;
Mondal et al. 2012; Diarra et al. 2017;
Meshram et al. 2018), with promising results.
To ensure the availability around the season and geographical locations, a
greater number of leaf meals should be evaluated in aquafeed. The current study
was intended to evaluate the potential of introducing mucuna
common carp pellets are enriched with leaf meal; there is a widespread cultured
Species of fish found throughout the world. The nutrition of the Mucuna bean (Mucuna
pruriens var. Use), grains legumes native to tropical regions, a similarity
can be found between soybeans and other legumes commonly found in agriculture,
as it includes the same levels of nutrients, such as protein, fat, minerals and
vitamins (Siddhuraju et al. 2000). According
to Duke (1981), its value as a source of nutritional protein for animal feeds
is well known, especially in underdeveloped nations. Cyprinus carpio L., sometimes known as common carp, is an
economically important freshwater fish species that accounts for 8% of global
finfish aquaculture production (FAO 2018). Under a variety of geographical,
meteorological and technical conditions, it is the world's third most farmed
species (Safari et al. 2016). Since its farming becomes common in
tropical areas, it's essential to evaluate cheaper feed resources like leaf
meals in the feed of this commercially important species.
Materials and
Methods
Experimental details and treatments
Leaf meal
preparation: M. bracteata DC. ex (Access no: 14825) leaves were
collected from Calicut, Kerala. Tap water was used to remove the dirt from the leaves
contaminants and obtain the lowest amount of moisture without altering the
profile of nutritional intake., Then it was dehydrated for two days in a shaded
place. Dried leaves were grounded with a mixer-grinder, kept in an airtight
container in ICAR-CIFEs feed lab.
Experimental diets:
Four
isonitrogenous and isocaloric diets (32% CP and 356 kcal DE/100 g) were
prepared by including Mucuna leaf meal (MLM) at 10, 20 and 30% in the diet,
intended to replace DORB at 33.3, 66.6 and 100%. DSBM, GNOC, MLM, WF, DORB and
CMC were weighed accurately (Table 1) and mixed to create a homogeneous
mixture. At 121°C, the dough was steam-cooked for 25 min. Oil, BHT, after the
dough had cooled; vitamin-mineral premix and choline chloride were combined. An
automatic pelletizer was used to pelletize the dough (SB Panchal Company).
After one day at room temperature, the pellets were dried under a fan, then for
one hour in a 40°C oven. After that, the pellets samples were allowed to dry
for one day at room temperature under a fan, following that, it was dried for
one hour using a 40°C oven, before being cooled and stored until needed at 4°C
in sealed containers.
Proximate analysis of leaf meal and diets
At the Fish Nutrition Laboratory, CIFE, dietary proximate analysis was
carried out utilizing conventional techniques (AOAC 1995). By removing the
weight before and after an overnight drying at 105°C, the moisture content was
calculated. The crude protein content % and crude lipid content % were determined
using Micro-Kjeldahl and Soxhlet extraction. A muffle
furnace kept at 450°C was used to ash the feed and ingredients and the crude
ash content (%) was estimated after weighing the ashes residue. Acid and alkali
digestion were used to calculate the crude fiber content (CF%). Nitrogen free
extract was computed by the formula:
NFE % =
100 [Crude protein (%) + Ether extract (%) + Crude fibre (%) + Total ash (%)].
Antinutritional factors (ANFs)
The Vaintraub and Lepteva
(1998) spectrophotometric approach were used to measure the phytic acid content
in MLM. According to Makkar et al. (2007)
total tannin was determined using a spectrophotometric approach. Day and
Underwood (1986) spectrophotometric method for estimating total oxalate was
used. The gravimetric method was used to determine the amount of alkaloids in
leaf meal. The saponin content of the leaf was
determined using a technique developed by (Hiai et al. 1976).
Experimental animal procurement
and acclimation
The fingerlings of C. carpio were sent to the ICAR-CIFE wet
laboratory facility in Mumbai, where two FRP circular tanks were with constant
aeration. A control diet was given to the fish for 21 days to acclimate them
(31 g crude lipid 100 g-1 and 6 g lipid 100 g-1).
Experimental design
The present study used a completely randomized
design. One hundred and eighty acclimated C.
carpio, in triplicates, fingerlings (average
bodyweight 6.06 - 0.08 g) were assigned to four experimental groups at random.
The feeding trial lasted 60 days and took place at the ICAR-wet CIFE's
laboratory in Mumbai, India, (tank measurements: 57%20139-1146_files/image002.png){kind=small}
36 47 cm, 150 L water volume, 175 L capacity).
Fish in each experimental tank were fed up to satiation using the respective
experimental diet two times a day (9 AM and 5 PM). Each experimental tank's
fecal matter was siphoned out and replaced with an equal volume of siphoned
water before the feeding for the next day began in the morning.
Estimation
of metabolic and oxidative stress enzymes
Following the completion of the experiment, each replicate had two fish
anesthetized with 50 l/L clove oil and the liver and muscle were dissected and
a mechanical homogenizer was used to prepare using a cold sucrose solution (0.25
M), homogenize the tissue. The whole
thing was done in an ice bath, and the homogenates were centrifuged for 10 min at
5000 rpm. The supernatants had been accumulated and stored in vials at 20°C.
The
activity of alanine aminotransferase (ALT) was tested similarly AST, for
example, with exception of the substrate. AST in muscle and liver was measured
and as defined by Wooten (1964), nanomoles of produced oxaloacetate/mg
protein/min (1964). Including the substrate, the action of alanine
aminotransferase (ALT) was measured Table 1: The experimental diets'
proximate content and the formulation was given to C. carpio fingerlings for 60 days
|
Ingredients composition
(g kg-1) |
Diets
(Treatments)1 |
|||
|
Control |
MLM10 |
MLM20 |
MLM30 |
|
|
DSBM 2 |
38 |
38 |
38 |
38 |
|
GNOC3 |
19.5 |
15 |
10.5 |
6 |
|
Wheat flour |
5.3 |
9.8 |
14.2 |
18.7 |
|
DORB4 |
30 |
20 |
10 |
0 |
|
MLM5 |
0 |
10 |
20 |
30 |
|
Soybean oil & fish oil
(1:1) |
4.2 |
4.2 |
4.2 |
4.2 |
|
Vitamin-mineral mix6
|
1.2 |
1.2 |
1.2 |
1.2 |
|
Choline chloride |
0.2 |
0.2 |
0.2 |
0.2 |
|
CMC7 |
1.5 |
1.5 |
1.5 |
1.5 |
|
BHT8 |
0.1 |
0.1 |
0.1 |
0.1 |
|
Proximate composition (dry matter basis) |
||||
|
Dry matter (%) |
90.33 |
90.87 |
91.07 |
90.86 |
|
Crude protein (%) |
32.08 |
31.99 |
32.09 |
31.94 |
|
Ether extract (%) |
6.05 |
6.02 |
6.04 |
6.02 |
|
Crude fiber
(%) |
7.14 |
7.25 |
7.34 |
7.74 |
|
Nitrogen free extract (%) |
46.34 |
46.23 |
45.72 |
43.99 |
|
Total ash (%) |
8.45 |
8.51 |
8.87 |
8.82 |
|
DE9 (%) |
368.54 |
367.54 |
365.19 |
358.44 |
|
P:E10 (%) |
87.04 |
87.03 |
87.87 |
89.11 |
The mean results of triplicates are used to calculate
proximate composition.
1) Control = 30% DORB and 0% MLM; MLM10, 10% MLM in
replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM in replacement of
66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in replacement of 100% DORB and 70.2%
GNOC 2) DSBM = De oiled soybean meal; (3) GNOC = Groundnut oil cake; (4) DORB De
oiled rice bran; (5) MLM = Mucuna bracteata
leaf meal (6) Composition of vitamin mineral mix (AGRUMIN 3503) (quantity kg-1)
Vitamin A, 7,00,000 IU; Vitamin D3, 7,00,000 IU; Vitamin E, 250 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, 1 g; Choline Chloride, 150 g; Mn, 27,000
mg; I, 325mg; Fe, 1,500 mg; Zn, 6,000 mg; Cu,1,000 mg; Co,150 mg; Lysine, 10 g;
Methionine, 10 g; Selenium, 125 mg; Vitamin C, 2,500 mg (7) CMC = Carboxymethyl
cellulose; (8) BHT = Butylated hydroxytoluene (9) DE (Kcal 100 g-1),
Digestible energy = {(kcal 100 g-1) = (%CP*4) + (%EE*9) + (%NFE*4)};
(10) P:E (mg protein kcal-1 DE), Protein to energy ratio (mg protein
kcal-1 DE) = (%CP*1000)/DE (kcal 100 g-1)
Table 2: Proximate composition and antinutritional factors of MLM
|
Particulars |
Mucuna leaf meal |
|
Proximate composition (%) |
|
|
Dry matter |
103.19 ± 0.13 |
|
CP |
28.12 ± 0.18 |
|
CL |
2.63 ± 0.05 |
|
CF |
17.69 ± 0.62 |
|
NFE |
44.79 ± 0.79 |
|
ASH |
18.76 ± 0.45 |
|
Gross energy (kcal/100g) |
370.61 ± 8.76 |
|
Antinutritional factor |
(mg/100 g) |
|
Total tannins |
21.16 ± 0.43 |
|
Alkaloid |
2.40 ± 0.01 |
|
Phytic acid |
13.76 ± 0.15 |
|
Saponin |
13.37 ± 0.44 |
Data represent mean ± S.E (n=6) MLM = M. bracteata
leaf meal; C.F = crude fiber; CL = crude lipid; CP =
crude protein; NFE = nitrogen-free extract; ASH = total ash content
correspondingly to the same AST. Nanomoles of pyruvate generated per
milligram of protein every minute was used to express ALT activity. Misra and Fridovich (1972) approach
was used to calculate the activity of the liver's superoxide dismutase (SOD).
The Action of superoxide dismutase could be calculated using the needed amount
of protein to reduce epinephrine autooxidation by 50% for one minute. The catalase
(CAT) activity was measured in nanomoles of H2O2
decomposed per minute per mg protein, according to Takahara
et al. (1960).
Blood, serum sampling and analysis
After five fish are being used in the experiment, chosen at random from
each of the tanks (15 per treatment) and anesthetized (clove powder, 200 mg L-1)
a 2 mL syringe to draw blood samples from the caudal vein. The serum was
separated by centrifuging the samples spun at 4000 rpm for 5 min using
non-heparinized tubes at 4°C. For the hematological indices experiment, Blood
was drawn into heparinized tubes and allowed to coagulate for 40 min at room
temperature. Leucocyte and erythrocyte count and hemoglobin concentrations were
all calculated using whole blood samples. The total amount of red blood cells
(RBCs) and white blood cells (WBCs) in the body (WBCs) were calculated using
the Jain (1976) and Carrol procedure. According to Rook and Dennis (1985), The
reduction of nitro blue tetrazolium was used to calculate the formation of
superoxide ions by leukocytes (NBT, Sigma-Aldrich Chemical, USA). Using a
Semi-auto Chemistry Analyzer, Erba Mannheim Mannheim kits, serum glucose, total protein, and albumin levels were calculated
using these methods (Rayto RT-9200, GmbH, Germany).
Albumin values were subtracted protein derived from whole serum to measure
globulin. Divide albumin measurements by globulin
values to get the albuminglobulin ratio.
Statistical analysis
The study used a complete randomized design. The mean and the data's
standard error were computed. At a 95 percent confidence level, the results
were subjected to a one-way analysis of variance using the Statistical Package
for Social Sciences (SPSS, version 16). Duncan's multiple range test was used to
measure the degree of significance between means.
Results
Nutrient and antinutrient profile
of MLM
MLM's nutritional and antinutrient profile is given in Table 2.
Phyto-chemical analysis of M. bracteata leaves showed that tannin content was 21.16 ± 0.43; phytate was
13.76 ± 0.15; alkaloid was 2.40 ± 0.01 and saponins was 13.37 ± 0.44.
Proximate composition of whole-body fish and
experimental diets
Table
1 shows biochemical dietary composition in experiments. During the experiment,
chemical constituents such as dry matter (90.3391.07), crude protein
(31.9432.09), ether extract (6.026.05), crude fiber (7.14-7.74) ash content
(8.458.87) and total carbohydrate (43.99-46.34) showed no significant (P > 0.05) alterations. Table 3 shows
entire fish carcass composition fed varied experimental diets. Table 3: Proximate analysis of the whole body of
several experimental groups of C. carpio
fingerlings (%wet weight basis)
|
Treatment1 |
Moisture |
Crude protein |
Ether extract |
Total
carbohydrate |
Total ash |
|
Control |
73.85ab
± 0.83 |
16.01cd
± 0.08 |
3.98 ± 0.17 |
3.72 ± 0.24 |
2.47 ± 0.15 |
|
LM15 |
74.59ab
± 0.61 |
15.54bc
± 0.38 |
3.83 ± 0.18 |
3.85 ± 0.28 |
2.23 ± 0.15 |
|
LM30 |
75.85c
± 0.59 |
14.51a
± 0.31 |
3.41 ± 0.21 |
3.95 ± 0.23 |
2.30 ± 0.13 |
|
CEE |
73.74ab
± 0.84 |
16.32cd
± 0.34 |
4.12 ± 0.16 |
3.42 ± 0.22 |
2.44 ± 0.26 |
All data are presented as Mean S.E. (n=6). The
difference between the mean values in the same column with different
superscripts is significant (P < 0.05)
1 Control, 30% DORB and 0% MLM; MLM10, 10% MLM in
replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM in replacement of
66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in replacement of 100% DORB and 70.2%
GNOC
Table 4: Protein
metabolic enzyme activities in C. carpio
fingerlings fed various experimental diets fish 60 days
|
Treatments1 |
Protein metabolic enzymes |
|||
|
AST2 |
ALT3 |
|||
|
Liver |
Muscle |
Liver |
Muscle |
|
|
Control |
1.28 ± 0.01 |
1.29 ± 0.11 |
2.76b
± 0.34 |
2.54b
± 0.02 |
|
MLM10 |
1.33 ± 0.09 |
1.36 ± 0.12 |
2.45b
± 0.18 |
2.32b
± 0.02 |
|
MLM20 |
1.49 ± 0.07 |
1.68 ± 0.14 |
3.72a
± 0.26 |
3.17a
± 0.37 |
|
MLM30 |
1.27 ± 0.03 |
1.28 ± 0.12 |
2.11b
± 0.26 |
1.95b
± 0.01 |
The data is shown as Mean Standard Error (n=6); the
difference between same-column mean values with distinct superscripts is
significant (P < 0.05)
1 Control, 30% DORB and 0% MLM; MLM10, 10% MLM in
replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM in replacement of
66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in replacement of 100% DORB and 70.2%
GNOC
2 AST,
Aspartate aminotransferase, the amount of oxaloacetate released is measured in
nanomoles. min-1 mg-1 protein at 37°C; 3ALT,
Alanine aminotransferase, nanomoles of sodium pyruvate released are used to
measure particular activity min-1 mg-1 protein at 37°C
Table 5: Hepatic
antioxidant enzyme activities in C. carpio fingerlings fed various experimental diets fish 60 days
|
Treatments1 |
SOD2 |
CAT3 |
|
Control |
9.06c ± 0.09 |
26.18b ± 0.15 |
|
MLM10 |
9.09c ± 0.06 |
24.94a ± 0.01 |
|
MLM20 |
9.45b ± 0.06 |
31.93d ± 0.04 |
|
MLM30 |
13.38a ± 0.23 |
29.13c ± 0.04 |
The data is shown as a mean SE (n=6); the difference
in mean values in the same column with different superscripts is considerable (P < 0.05). 1Control, 30% DORB and 0%
MLM; MLM10, 10% MLM in replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM
in replacement of 66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in replacement of
100% DORB and 70.2% GNOC
2SOD, Superoxide
dismutase, Inhibition of epinephrine auto-oxidation by 50% is the measure of
specific action mg-1 protein min-1; 3CAT,
Catalase, nanomoles are used to measure particular activity H2O2
decomposed min-1 mg-1 protein
During experiment, chemical constituents such as moisture (73.7475.85)
and crude protein (14.5116.32) showed differences that are significant (P < 0.05). The crude lipid (3.414.12),
ash content (2.232.47) and total carbohydrate (3.42-4.12) showed no
significant (p > 0.05) shifts.
Protein metabolic enzyme and oxidative stress enzyme
The ALT activity of muscle and liver (Table 4) fluctuated considerably
between dietary groups (P < 0.05).
Between experimental groups, there was no significant difference in AST
activity in muscle or liver (P > 0.05).
Hepatic SOD and catalase behavior (Table 5) a significant difference between
the experimental group (P < 0.05).
Hepatic SOD activity was higher in MLM30% fed fish (P < 0.05) compared in control fish, while fish given only 20%
dietary MLM had considerably higher liver catalase (CAT) activity compared to
the control group (P < 0.05).
Haemato-immunological profile
MLM leaf meal had a major impact on the overall RBC count. A rise in RBC
count was observed as the MLM level increased (Table 6). The count of RBCs and
WBCs in the control group and The MLM10% groups were statistically comparable (P > 0.05). The NBT values differ
significantly (P > 0.05) among the
treatments. When MLM was used, the NBT value increased dramatically in
comparison to a control group. The treatment with MLM30 had a higher NBT value.
Serum proteins
Table 7
shows albumin, globulin and total protein levels in the blood, for various
experimental groups. The control group had the highest albumin blood
albumin-globulin ratio and concentration, which was not statistically different
(P > 0.05) from other groups given
leaf meal but was higher in comparison to other groups. The MLM10 % group had
the highest serum globulin levels; this was statistically different from the
other groups (P > 0.05). The serum
glucose levels (Table 7) a significant difference among the treatments (P > 0.05). MLM participation in 10%
and 20% resulted in a substantial reduction in serum glucose levels as in
comparison to a control group, while participation in 30% resulted in an
increase in serum glucose level but the increase was not significant.
Discussion
This the purpose
of the research was to see if raw Mucuna DORB could be replaced in the diet
with leaf meal of common carp. However, there have been no studies on M. bracteata Table
6: Hematological
and haemato-immunological parameters of C. carpio for
60 days, fingerlings were fed various experimental diets
|
Treatments1 |
Hemoglobin (g dL-1) |
TEC2 (x106 cmm-1) |
TLC3 (Χ104 cmm-1) |
NBT4 (OD 620 nm) |
|
Control |
7.25c ± 0 .05 |
2.00b ± 0.03 |
14.05b ± 0.43 |
0.55b ± 0.01 |
|
MLM10 |
8.08b ± 0.12 |
1.96b ± 0.02 |
14.03b ± 0.08 |
0.58b ± 0.02 |
|
MLM20 |
8.93a ± 0.04 |
2.21a ± 0.01 |
12.49c ± 0.32 |
0.65a ± 0.01 |
|
MLM30 |
7.46c ± 0.04 |
1.89c ± 0.02 |
15.69b ± 0.43 |
0.68a ± 0.01 |
The data is shown SE as Mean (n=6); the difference in mean values in the
same column with different superscripts is considerable (P < 0.05)
1Control, 30% DORB and 0% MLM; MLM10, 10% MLM in
replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM in
replacement of 66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in
replacement of 100% DORB and 70.2% GNOC 2TEC, Total erythrocyte
count; 3TLC, Total leucocyte count; 4NBT, Nitoblue tetrazolium
Table 7: Haemato-biochemical characteristics in fingerlings
of C. carpio
fed various experimental diets for 60 days
|
Treatments1 |
Serum TP2 (g dL-1) |
Serum Alb3 (g dL-1) |
Serum Glob4 (g dL-1) |
Serum A:G5 |
Serum Glu6 (mg dL-1) |
|
Control |
3.03 ± 0.24 |
1.37 ± 0.23 |
1.57c ± 0.01 |
0.87 ± 0.12 |
82.15a ± 0.33 |
|
MLM10 |
2.93 ± 0.07 |
1.12 ± 0.02 |
1.94b ± 0.06 |
0.57 ± 0.01 |
80.43b ± 0.54 |
|
MLM20 |
3.01 ± 0.04 |
1.16 ± 0.03 |
1.54c ± 0.03 |
0.75 ± 0.02 |
79.42b ± 0.31 |
|
MLM30 |
3.08 ± 0.13 |
1.20 ± 0.07 |
1.74a ± 0.05 |
0.68 ± 0.02 |
83.09a ± 0.37 |
The data is shown as Mean Standard Error (n=6); the difference between
mean values in the same column with different superscripts is significant (P < 0.05). 1Control, 30% DORB and 0%
MLM; MLM10, 10% MLM in replacement of 33.3% DORB and 23.4% GNOC; MLM20, 20% MLM
in replacement of 66.6% DORB and 46.8% GNOC; MLM30, 30% MLM in replacement of
100% DORB and 70.2% GNOC
2TP, Total protein; 3Alb, Albumin; 4Glob,
Globulin; 5A:G, Albumin to globulin ratio; 6Glu, Glucose
leaf meal has been used as a DORB
alternative in fish diets. M. bracteata leaf meal is
used in fish diets has never been documented., but there is information on
other leaf meals. Leaf meals are inexpensive, easily accessible, and have no
direct competitors in the livestock feed industry. Leaf meals can be
utilized in the C. carpio diet as an
alternate aquafeed component, several prior studies have found (Meshram et al. 2018; Anand et al. 2019; Ahmad
et al. 2019; Maiti et al. 2019; Jayant et
al. 2020; Rani et al. 2020; Sahoo et al. 2020). The major
challenge in limiting the use of alternative feed sources of plant origin is
its fish acceptability, which is typically linked to the diet's palatability
(Rodriguez and Perston 1996). The majority
of tannins, oxalates, phytic acid, alkaloids and other anti-nutritional
substances and other anti-nutritional factors can be found in plant substances
(NRC 1993), which obstruct nutrition use and have detrimental consequences for
animal growth and other physiological functions, including fish. Total tannins,
phytic acid, saponins, and total alkaloids were discovered in MLM. For the
anti-nutrient content of MLM, there are no quantitative reports available.
Anand et al. (2019) reported that C. carpio
fed a 15% raw Sesbania leaf meal-based diet had worse development efficiency,
most likely due to increased saponin levels. Green pea meal could be used to
substitute 10% of fish meal in barramundi and Lates calcarifer feed without
affecting growth or nutrient utilization (Ganzon-Naret
2013). Puycha et al. (2017) found that higher inclusion of
moringa, Swai catfish were fed 150 to 200 grams of Moringa oleifera leaf meal per kilogram
of body weight, Pangasius bocourti, possibly due to its high phytate content
(0.4%). Dietary tannin and phytic acid concentrations of 0.5-2.0% and 0.5-0.6%,
respectively, were found to reduce C. carpio growth performance (Hossain and Jauncey 1990). The
experimental diets using mucuna seed meal
demonstrated a high acceptability and no essential amino acid (EAA) deficits (Siddhuraju and Becker 2001). Sweet potato leaf meal, raw (Meshram et al. 2018) and Hygrophila spinosa leaf meal (Maiti et al.
2019) were found to DORB should be fully replaced (30% incorporation)
within the feed of Labeo rohita.
Proximate composition of the M.
bracteata
leaves indicated that the leaves of M. bracteata
are relatively rich in crude protein (28.12 ± 0.18). Janardhanan and Lakshmanan (1985) discovered that the seeds
of M. pruriens var. utilize to have a high crude
protein content (26.2529.6%). According to Jambunathan
and Singh (1980), the crude protein value of pulses, pigeon pea, black gram,
red gram, chickpea and green gram are often consumed, appears to be lower than mucuna. The protein level, in
particular, tends to compare favorably to recorded levels of other traditional
protein sources, especially those of plant origin (Nwokolo
and Oji 1985; Fasuyi et al. 2008). Mucuna
meal-based diets resulted in the highest carcass moisture and crude protein
levels and contain the least amount of lipids and energy. Surprisingly, the
difference was insignificant in carcass moisture, protein, ash, or calorie
value between the treatment diets. This may be due to the experimental diets
being isonitrogenous and isolipidic (Yengkokpam et al. 2013; Fawole
et al. 2016; Garg et al. 2019).
When fish are stressed and have an energy shortfall, existing amino
acids are converted to a different amino acid by transaminases, which are
subsequently reduced for the generation of energy from keto acid
gluconeogenesis (Silva and Anderson 1995; Chatterjee et al. 2006). As a
result, the availability of nonproteins, the need for amino acids in energy
synthesis can be reduced by using alternative energy sources. In this way, ALT
and AST help fish growth by serving energy metabolism as a link between
protein, carbohydrates, and fats (Shamna et al.
2015). As a result, the activity of the AST and ALT enzymes can be useful
markers in terms of amino acid metabolism in fish (Lin and Luo 2011; Jiang et
al. 2015). Higher ALT and AST activities in the MLM20 group in this study
suggested that fish could make nonprotein energy, particularly carbohydrate,
available at an optimal level, thus sparing amino acids facilitate the
production of body proteins, resulting in increased fish development.
When tissue free radicals are abnormally strong, animals suffer from
oxidative stress (Sies and Groot 1992). Antioxidant defense mechanisms in all
organisms protect them from ROS-mediated cellular and nutrient damage, with CAT
and SOD playing a key role in lowering ROS levels in tissues. Catalase and
peroxidase are enzymes that convert superoxide ions to water, hence an increase
in their activity is thought to boost antioxidant defenses (Dawood et al.
2017b; Abdel-Tawwab and Monier
2018; Hoseinifar et al. 2018; Doan et al.
2019). SOD converts the poisonous superoxide anions to hydrogen peroxides,
which are then degraded by CAT into oxygen and water. SOD and CAT movements are
thus increased during oxidative stress to neutralize ROS. As a result, when
there is oxidative stress, SOD and CAT activity both rise. In this work, higher
SOD and CAT levels in MLM groups that have been fed demonstrated oxidative
stress caused by ANF in fish.
Hematological
parameters in fish have been demonstrated to be improved by extracts from
plants (Reverter et al. 2014; Dawood et al.
2017a). As the amount of Mucuna leaf
meal as a dietary supplement increased, the number of red blood cells
increased. The rise in WBC in common carp fed MLM enriched meals
was found, particularly in this study, the MLM20 supplemented diet was used,
which could be due to MLM's immunostimulatory properties. WBC numbers have
phagocytic activity as well as biomarkers for immunological function. According
to Valenzuela-Grijalva et al. (2017), immunostimulatory activity in
animals is one of the biological effects of phytogenic feed additives. Between the control and treatment
groups, there was a substantial difference in RBC, Hb and WBC (P > 0.05). In this analysis, indices
like RBC and Hb in C. carpio through
the different dietary treatments indicate that the MLM diet supports or does
not obstruct regular functioning hemopoiesis processes. The higher RBC levels
noticed in fish-fed individuals' MLM20 diets suggest that the dietary protein
content is higher and that MLM20 improved blood quality. Increased RBC values
were related to high-quality dietary protein and disease-free livestock,
according to Hackbarth et al. (1983). The
measurement of oxidative radical generation is critical for evaluating the
stimulation of cellular defense in fish. Some immune
cells, such as neutrophils and macrophages, use oxygen during this process and
produce reactive oxygen species (ROS), which are harmful to pathogenic germs
(Srivastava and Pandey 2015). In a reduction reaction, these ROS interact with
nitro blue tetrazolium (NBT), showing whether or not oxidative radical
generation has risen.
Conclusion
Finally, the findings imply that feeding common carp a diet containing
20% MLM for 60 days may be sufficient to increase immunological parameters,
protein metabolic enzymes, and antioxidant status. As a result, Mucuna leaf
meal can be used to substitute regular rice bran that has been de-oiled in common
carp feed. Mucuna leaf meal offers potential as a rice bran replacer, according
to the findings of this study. The nutritional quality of M. bracteata leaf meal, on the
other hand, was adequate, resulting in metabolic and antioxidant enzyme
activities. However, the inclusion of MLM at 20% exhibited higher better
physio-metabolic responses in fish than DORB. As a result, more to analyze and
understand the effects of varying quantities of dietary mucuna
leaf meal on a variety of finfish species, more research is needed.
Acknowledgments
The authors express sincere thanks to the Director/Vice-Chancellor,
ICAR-CIFE, Mumbai, India, for as well as the necessary infrastructure for the
current experiment and lab analysis. The institutional Ph. D. fellowship, as
well as ICAR (Indian Council of Agricultural Research) to the first author, is
gratefully acknowledged.
Author Contributions
Ashutosh Dharmendra Deo and Narottam Prasad Sahu planned experiments, Tincy
Varghese and Hafeef Roshan interpreted the results, Hafeef Roshan made the review and genuinely broke down the
information and made outlines.
Conflict of Interest
The authors do not have any
conflict of interest to declare.
Data Availability
The appropriate author will
be able to provide data upon request
Ethical Approval
The institutional ethical
committee gave its approval to all of the ethical rules that were followed.
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