Chong M. Lee 1, Peizhi Lian 1, David Bengtson 2, Nick King 3 and George Nardi 4 1. Dept. of Nutrition and Food Sciences 2. Dept. of Fisheries, Animal and Veterinary Science University of Rhode Island 3. Skretting, Vancouver, BC 4. GreatBay Aquaculture, Portsmouth, New Hampshire

Commercial scale production of squid hydrolysate 330 gal capacity 86% M 73-78% M Yield: 47-54% Vibrating screen In-line viscometer Evaporator Hydrolysis Raw squid byproduct hydrolysate

Proximate composition of squid hydrolysate (concentrated – 76% moisture) MoistureProteinLipidAsh 76 %18.63 %3.30 %2.19 % Squid hydrolysate contains 11.16% EPA and 24.45% DHA, while salmon oil contains 8.65% EPA and 10.67% DHA (on an oil weight basis).

Ingredients Amount (% dry wt basis) Squid hydrolysate73.33 Salmon oil9.54 Lecithin3.01 Vitamin- premix0.44 Mineral premix2.01 Starch5.02 Yeast4.02 Algae (spirulina: chlorella)2.64 Proximate composition Protein64.66 Lipid18.72 Carbohydrate7.40 Ash9.21 Energy (MJ/Kg)19.12 Composition of squid hydrolysate-based basal microdiet

Pilot plant production of microparticulate feed Grinding homogenizng drum drying sieving

(DHA) (EPA) Fatty acid profiles of microdiets *The 100 g basal squid hydrolysate diet will provide 2.00 g EPA and 3.60 g DHA based on EPA/DHA distribution (% fatty acid/oil).

Field feeding trial on Atlantic cod larvae: weaning procedure n 0.25 million of cod larvae/ production tank (5 m 3 ). n One tank for squid hydrolysate(SH)-larval diet n Six tanks (control group) for a premium commercial diet n Cod larvae upon hatch were on - initially rotifer - the combination of rotifer and Artemia - co-feeding of Artemia and microdiet Weaning 1 week later with gradual removal of Artemia, the fish were kept on the SH microdiet for another 2 weeks. (The trial was conducted at the GreatBay Aquaculture in Portsmouth, NH)

Survival n 70-75% of the fish on the SH microdiet survived through the weaning period, which is considered excellent (in the same range for the control group - the premium commercial diet) n Overall, there was no real difference in survival among the production tanks during weaning.

Swimming behavior n Different behavior between the fish fed SH diet and the rest. n The SH fish had a lighter color. A darker color is often associated with stress. n The SH fish were very responsive as a sign of good health being more uniform in size. This has very significant ramifications as it relates to cannibalism and grading. n The fish were swimming together in uniform manner and appeared to be in motion more so than those in the control diet tanks.

Tank hygiene n The SH diet was rated better than the premium control diet. n The SH diet appeared to stay very stable in the water without leaching. Leaching tends to cause foam on the surface (which was a problem with the control diet).

* GM: the premium commercial diet ** Salinity at 65 ppt for 60 min (CSI: cumulative stress index) The number of dead larvae counted every 3 min. At the end of 60 min, the % cumulative mortality was used as a Cumulative Stress Index (CSI-60). Indication of "condition“ after treatments, stress resistance, and screening DPH / / / / / / / Salinity stress test for cod larvae (48 dph) Total length (mm)

Microdiets from SH with different hydrolysis time on growth performance of Atlantic cod larvae (50 dph*) in 42 days feeding SH 2: 2 h hydrolysis at 55C, followed by 30 min at 75C SH 1: 1 h hydrolysis SH 2-1: fed 2 h hydrolysate diet for 21 d followed by 1 h diet for 21 d IBL: initial body length; BW: body wt; BCF: body condition factor; SGR: specific growth rate * 30 dph and 20 d acclimation

SDS-PAGE profiles of squid by-product during hydrolysis Molecular marker (kDa) VISCOSITY DEGREE OF HYDROLYSIS

Surivial, weight, length and specific growth rate of summer flounder larvae (Paralichthys dentatus) after 22-day

Additional studies to be considered n Feeding trials on additional species n Hydrolysate from finfish-squid combination n Squid hydrolysate for broodstock nutrition n Feed performance test in plant protein-based aquaculture feeds