EnzoMeal Evaluation for Commercial
Production of Yellow Perch
Presented by Dr. Vikas Kumar from
Kentucky State University
Presented at World Congress on
Industrial Biotechnology 2016
Aim
Control
(Fish meal)
Growth and health of
Yellow perch
Soybean meal
(+/- enzymes)
8/11/2016
EnzoMeal
(+/- enzymes)
2
Experimental design
Yellow perch (297; av wt.= 11 g) - nine
groups with three replicates
Flow through system (18 inch tank)
Water parameters: optimum range
(Water temp: 19 – 24°C, pH: 6.5 – 7.6,
DO: 6.5 – 7.7)
Potential candidate for
aquaculture in North America
Experimental period: 10 weeks
Fish fed (hand feeding) 2-3% body
mass – 4 times in a day
Diets: nine diets (Isonitrogenous and isolipidic)





Crude protein – 38%,
Crude lipid – 12%,
Vitamin premix - 3%,
Mineral premix - 4%
TiO2 – 1%
3
Composition of the experimental diets (% dry matter basis of ingredients), containing
roasted soybean meal (SBM) and EnzoMeal (EZM)
Experimental diets
Diet 3
Diet 4
Diet 5
Diet 6
Diet 7
Diet 8
Diet 9
Control SBM50
SBM100
EZM100
48.0
24.0
-
SBM100 +
Enzyme
-
EZM50
Fish meal
SBM50 +
Enzyme
24.0
24.0
-
EZM50 +
Enzyme
24.0
EZM100 +
Enzyme
-
Wheat meal
34.8
21.9
9.0
21.9
9
28.4
21.8
28.4
21.8
Soybean meal
-
32.5
65.0
32.5
65.0
-
-
-
-
EnzoMeal
-
-
-
-
-
26.9
53.8
26.9
53.8
Soy protein isolate
-
1.6
2.8
1.6
2.8
0.7
1.2
0.7
1.2
Menhaden oil
5.0
7.8
10.7
7.8
10.7
7.8
10.7
7.8
10.7
Soybean oil
5.0
5.0
5.0
5.0
5.0
5.0
5.0
5.0
5.0
Vitamin premix
3
3.0
3.0
3.0
3.0
3.0
3.0
3.0
3.0
Mineral premix
4.0
4.0
4.0
4.0
4.0
4.0
4.0
4.0
4.0
TiO2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
Methionine
-
-
0.3
-
0.3
-
0.3
-
0.3
Cocktail enzyme
-
-
-
Yes
Yes
-
-
Yes
Yes
Total
100
100
100
100
100
100
100
100
100
Ingredients
Diet 1
Diet 2
Feeds were extruded at 45-50 oC to maintain the cocktail enzymes
activity in the diets and these diets were dried at 45-50 oC for 6 h
4
Growth performance (10 weeks)
Treatment
Initial weight (g) Final weight (g)
Control (Fish meal) 10.96 ± 0.06
SBM50
11.04 ± 0.07
39.96 ± 2.42ab
SBM50+Enzymes
10.91 ± 0.25
36.90 ± 8.88bc
SBM100
10.98 ± 0.06
22.04 ± 1.65e
SBM100+Enzymes
10.99 ± 0.07
22.51 ± 2.54e
EZM50
11.16 ± 0.11
40.02 ± 0.63ab
EZM50+Enzymes
11.06 ± 0.14
45.38 ± 1.70a
EZM100
10.71 ± 0.38
24.91 ± 0.85e
EZM100+Enzymes
11.18 ± 0.20
27.89 ± 1.71de
33.13 ± 2.30cd
Values are mean (n = 3) ± standard deviation.
Mean values in the same column with different superscript differ significantly (P < 0.05).
5
Growth performance (10 weeks)
Body weight gain (%) = 100*[Final body mass (g) – Initial body mass (g)]/ Initial body mass (g)
Values are mean (n = 3) ± standard deviation.
Mean values in the different bars with different superscript differ significantly (P < 0.05).
6
Nutrient utilization
Values are mean (n = 3) ± standard deviation.
Mean values with different superscript differ significantly (P < 0.05).
7
Nutrient utilization
Values are mean (n = 3) ± standard deviation.
Mean values with different superscript differ significantly (P < 0.05).
8
Nutrient utilization
Protein productive value (PPV) = Retention of protein in fish body from feed
Apparent lipid conversion (ALC) = Retention of lipid in fish body from feed
Values are mean (n = 3) ± standard deviation.
Mean values with different superscript differ significantly (P < 0.05).
9
Activities of digestive enzymes
Amylase
Protease
Lipase
Treatment
Pyloric caeca
Intestine
Pyloric caeca
Intestine
Pyloric caeca
Intestine
Control
4.88 ± 1.6ab
7.17 ± 1.9
42 ± 7.3a
57 ± 7.1a
8.00 ± 2.2
19 ± 2.2a
SBM50
5.42 ± 2.0ab
9.55 ± 1.3
31 ± 6.9ab
45 ± 6.1ab
6.48 ± 1.8
16 ± 1.8ab
SBM50 + Enzyme
4.30 ± 1.4ab
9.69 ± 4.6
25 ± 2.8bc
38 ± 2.8b
6.81 ± 1.3
18 ± 1.3 ab
SBM100
3.43 ± 0.7ab
5.61 ± 4.7
21 ± 5.3c
35 ± 5.3b
5.04 ± 2.1
13 ± 2.1 ab
SBM100 + Enzyme
2.59 ± 0.6b
6.76 ± 3.7
22 ± 3.1c
36 ± 3.1b
5.72 ± 3.5
16 ± 3.5 ab
EZM50
4.38 ± 0.7ab
7.48 ± 0.6
41 ± 3.2a
55 ± 3.2a
7.92 ± 2.3
18 ± 2.3 ab
EZM50 + Enzyme
6.64 ± 1.2a
13.22 ± 1.0
41 ± 10.7ab
55 ± 10.7a
8.66 ± 2.3
18 ± 2.3 ab
EZM100
3.77 ± 1.3ab
9.04 ± 2.0
18 ± 3.2c
32 ± 3.2b
4.15 ± 0.6
12 ± 0.6 b
EZM100 + Enzyme
3.91 ± 1.7ab
8.07 ± 3.3
24 ± 2.1c
36 ± 3.8b
6.74 ± 2.2
13 ± 2.2 ab
Values are mean (n = 3) ± standard deviation. Mean values with different superscript differ significantly (P < 0.05).
Amylase (nmole/min/mL) - One unit of amylase is the amount of amylase that cleaves ethylidene-pNP-G7 to generate 1.0 μmole of
p-nitrophenol per minute at 25 °C.
Protease (nmol/min/ml = mU/ml) - Unit Definition: One unit is defined as the amount of protease that cleaves the substrate, to yield an amount of fluorescence
equivalent to 1.0 nmol of unquenched FITC per minute at 25°C.
Lipase (nmole/min/mL = milliunit/mL) - One unit of Lipase is the amount of enzyme that will generate 1.0 μmole of glycerol from triglycerides per minute at 37 °C.
10
Metabolic enzymes
(protein metabolism)
Values are mean (n = 3) ± standard deviation. Mean values with different superscript differ significantly (P < 0.05).
ALT (U/ml) U- One unit is defined as the amount of enzyme that will cause the oxidation of 1.0 μmol of NADH to NAD+ per minute at 370C.
One unit of AST is the amount of enzyme that will generate 1.0 mmole of glutamate per minute at pH 8.0 at 37 °C.
11
Antioxidant enzymes
Values are mean (n = 3) ± standard deviation. Mean values with different superscript differ significantly (P < 0.05).
Superoxide dismutase (inhibition rate %) – production of water-soluble formazan dye upon reduction with a superoxide anion
Catalase (nmol/min/ml) – 1 U - the amount of enzyme that will cause the formation of 1.0 nmol of formaldehyde per minute at 250C.
Glutathione peroxidase: 1 U - the amount of enzyme that will cause the oxidation of 1.0 μmol of NADPH to NADP+ under the assay kit
12
condition per minute at 250C.