Development of a feed with EnzoMeal for commercial production of Pacific white shrimp (Litopenaeus vannamei) Presented by Dr. Vikas Kumar from Kentucky State University Presented at World Congress on Industrial Biotechnology 2016
Aquaculture is the fastestgrowing food sector for over 25 years, with an APR of 8.1% per year, compared with 0.62% for capture fisheries and 2.6% for agricultural meat production (Source: FAO, 2014) Global Aquaculture Production and Value from 2000-2014 2
Pacific White Shrimp (Litopenaeus vannamei) Most successful globally produced marine crustacean 72% of world production of all farmed marine shrimp Asia produced 78% of the 2.5 million tons
Aim and Objectives The overall aim of the present proposed project is to increase the usage of EnzoMeal (EZM) in feed for commercial shrimp production. Objective 1: Define the best feed formulation to incorporate maximum amount of EZM in shrimp diet; Objective 2: Evaluate the nutritional value of EZM based diets vs. fishmeal based feed for shrimp
Diet 4 (EZM 100 ): 100% FM protein replaced by EZM Materials and Methods Analyzed nutritional composition of raw ingredients using portable handheld probe device -Thermoscientific microphazir AG Obtained proximate composition (protein, lipid, fiber and ash) of each feed ingredients Shrimp feed were formulated according to the nutrient requirement of shrimp Crude protein 38% and lipid - 8% Diet 1 (control): 30% fish meal (FM) Diet 2 (EZM 33 ): 33% FM protein replaced by EnzoMeal (EZM) Diet 3 (EZM 66 ): 66% FM protein replaced by EZM
Shrimp Diet Formulations (g/100 g) Ingredient Control EZM 33 EZM 66 EZM 100 Fish meal 30.00 20.00 10.00 0.00 EnzoMeal 0.00 11.30 22.60 33.90 Chicken byproduct meal 10.00 10.00 10.00 10.00 Squid meal 2.00 2.00 2.00 2.00 Wheat meal 15.00 14.80 12.00 11.00 Corn distillers dried solubles 37.65 34.45 34.45 32.45 Wheat glutein 1.00 2.00 2.50 3.20 Fish oil 0.00 1.10 2.10 3.10 Dicalcium phosphate 1.50 1.50 1.50 1.50 Magnesium Sulfate 0.60 0.60 0.60 0.60 Potassium Phosphate 0.80 0.80 0.80 0.80 Lecithin 0.50 0.50 0.50 0.50 Vitamin premix 0.40 0.40 0.40 0.40 Mineral mix 0.10 0.10 0.10 0.10 Vitamin C (Stay C) 0.10 0.10 0.10 0.10 Cholesterol 0.20 0.20 0.20 0.20 Choline chloride 0.15 0.15 0.15 0.15 Total 100 100 100 100
Manufacturing the shrimp feed By commercial extruder at EnzoMeal
Members - for feed extrusion at Alltech Winchester, Kentucky
Experimental set up Indoor salt water recirculatory system (RAS) Salinity maintained at 27-28 ppt and Temp. 28 o C Total 12 tanks, each tank volume 1000 liter Stocked 200 shrimp (Av wt. 3.2 g) per tank
Water quality Measured daily: Hydrolab Quanta Water Quality Monitoring System and Spectrophotometer Dissolved oxygen Salinity and ph Temperature Alkalinity and Ammonia Statistical Analysis One-way ANOVA - Tukey s studentized range (honestly significant difference) test Statistical Analysis System version 9.3 (SAS Institute, Cary, NC, USA) Differences among mean - considered significant at the P<0.05 probability level
Sampling Weighing Every four weeks Experimental period: 12 weeks At harvest: Shrimp was anesthetized Gas Chromatography Shrimp tails (muscle) and hepatopancreas Proximate analysis feed and shrimp muscle Fatty acids - Shrimp muscle Hemolymph Parameters Thermo scientific microphazir AG IDEXX VetTest 8008 Physiological parameters: Hemolymph glucose Total protein, albumin, and globulin 11
Growth performance Feeding trial - 12 weeks Survival rate: 82 85%, no significant difference (P > 0.05) Initial weight (g) Final weight (g) Control (Fishmeal) 3.25 ± 0.03 25.5 ± 1.51 a EZM 33 3.24 ±0.04 24.1 ± 0.55 ab EZM 66 3.24 ± 0.03 22.2 ± 1.47 b EZM 100 3.23 ± 0.03 22.5 ± 1.18 ab Control: 30% fish meal (FM) EZM 33 : 33% FM protein replaced by EZM EZM 66 : 66% FM protein replaced by EZM EZM 100 : 100% FM protein replaced by EZM Values are mean (n = 3) ± standard deviation. Mean with different superscript differ significantly (P < 0.05). 12
Growth Performance Body mass gain (%) 600.00 400.00 200.00 No significant difference among the groups (P > 0.05). Similar growth performance of EZM fed groups compared to fishmeal group. 0.00 Control EZM33 EZM66 EZM100 Values are mean (n = 3) ± standard deviation. Mean with different superscript differ significantly (P < 0.05). 13
Nutrient Utilization Feed conversion ratio (FCR) = Total feed fed (g)/ total body mass gain (g) Protein efficiency ratio (PER) = Body mass gain(g)/ Total protein fed(g) No significant difference among the groups (P > 0.05). Similar performance of EZM fed groups compared to fishmeal group. Control EZM33 EZM66 EZM100 14
Treatment Urea Nitrogen (mg/dl) Health Parameters Hematological parameters Creatinine (mg/dl) Total Bilirubin (mg/dl) Glutathione (U/L) Control 3.3 ± 0.3 0.83 ± 0.4 3.0 ± 1.2 485 ± 127 EZM 33% 2.3 ± 0.3 1.00 ± 0.4 3.3 ± 0.5 512 ± 186 EZM 66% 2.7 ± 0.3 0.63 ± 0.1 4.1 ± 0.4 587 ± 46 EZM 100% 2.0 ± 0.0 0.57 ± 0.1 3.5 ± 1.3 583 ± 196 Pr > F 0.144 0.774 0.381 0.979 Values are mean (n = 3) ± standard deviation Mean values in the same column with different superscript differ significantly (P < 0.05) 15
Health Parameters Concentration of protein in haemolymph The concentration of total protein in haemolymph is used as a basic index for health and nutritional status in shrimp Control EZM33 EZM66 EZM100 Values are mean (n = 3) ± standard deviation. Mean with different superscript differ significantly (P < 0.05). 16
Health Parameters Concentration of glucose concentration in haemolymph It is considered as an indicator of stress parameter Overall there is no significant difference for health related parameters: Animals are normal and healthy Control EZM33 EZM66 EZM100 Values are mean (n = 3) ± standard deviation. Mean with different superscript differ significantly (P < 0.05). 17
Conclusions EnzoMeal: comparable to fish meal and better than soybean meal EnzoMeal can replace 50-75% fish meal protein in fish whereas 100% replacement in shrimp Promising new ingredient for the growing aqua feed industry 18
Acknowledgement Thank You! 19